Science method

Cytology - Science method

Explore the latest questions and answers in Cytology, and find Cytology experts.
Questions related to Cytology
  • asked a question related to Cytology
Question
1 answer
I’m looking for recommendations on the best fixative for preserving vaginal cytology swabs collected from wild polar bears in extremely cold field conditions. Preparing slides on-site isn’t an option, so we need a fixative that allows the swab to be immersed and preserves the cells for PAP staining at a later date—potentially up to three months post-collection.
Given the freezing temperatures, our options for fixatives are somewhat limited. We are considering 95% ethanol, knowing it may cause some cell shrinkage, but we’re open to better alternatives that would maintain cellular integrity for later staining.
Has anyone worked with long-term storage of cytology swabs in harsh conditions? Are there fixatives or techniques you would recommend to minimize cellular distortion while ensuring preservation?
Any insights or references would be greatly appreciated!
Relevant answer
Answer
Dear Louisa A Rispoli,
The standard fixative for PAP is ethanol, however in such circumstances you can use 4.5% neutral buffered formalin and keep the vehicle moist during sampling. Later you can wash gently with water to remove the formalin and go forward with the staining procedures using PAP.
You can also use Giemsa stain if you have difficulties with it or you would like to compare the cellular elements .
Best wishes
  • asked a question related to Cytology
Question
4 answers
Hi everyone, In the diagnostic workflow of the test, we often need to reuse cytology slides, which requires removing the coverslip previously mounted over the sample. Currently, we use xylene, and the slides are left immersed for up to 72 hours until the coverslip detaches. However, we are looking for a faster and more efficient method to optimize our routine. Has anyone had experience with alternative solvents or techniques that are more effective for removing the coverslip and the mounting medium (typically resin or Canada balsam)? Any suggestions based on practical experience or literature references would be greatly appreciated.
Relevant answer
Answer
Klaus Berkefeld Thank you so much for the additional details, Klaus — that’s very helpful! Indeed, your point about the variability of mounting media is important, and I appreciate your clarification about not using heat and letting the slides sit overnight. We’ll definitely consider testing this approach with a few slides, including scoring the coverslip beforehand, and see how it performs in our context. Thanks again for your valuable insights and your time!
  • asked a question related to Cytology
Question
12 answers
I am planning to expand my business and hope to learn some industry information from you in your area. Or share some resources and explore potential cooperation opportunities.
#Medicine #Biotechnology #Doctor#Research
Relevant answer
Answer
Yes, I am interested to work with you.
  • asked a question related to Cytology
Question
3 answers
Liquid cytology is more sensitive and specific than conventional cytology. Its procedure is easier and less time consuming. The image showing a case of adenocarcinoma.
Relevant answer
Answer
I would argue, it could be lab and staff-dependant
  • asked a question related to Cytology
Question
3 answers
Hi, I am Sebastian, a bachelor of Veterinary Medicine from Lima, Peru. My main interests are internal medicine, emergency care, evidence-based medicine, cytology, and other topics related to small animals. For this reason, I am searching for online courses to develop my skills as a veterinarian.
I would appreciate it if you could recommend some online courses that offer this type of training, preferably from universities. Thank you.
I hope this helps! Let me know if you have any more questions.
Relevant answer
Answer
Everybody can find every subject totally free of tuition fees in YOUTUBE videos. If anybody really wishes to learn !?
  • asked a question related to Cytology
Question
3 answers
I am doing an analysis of a dataset. And I want to find out if cytological features can predict/correlate well with acid fast bacilli positivity/negativity. I know such work has been but I am considering using multiple logistic regression analysis.
Predictor = cytology
Outcome = Positive AFB
I am not sure if multiple logistic regression analysis tool is the best statistical tool to use for this prediction model. I would appreciate any feedback or comments.
Relevant answer
Answer
I am new to R so I have a steep learning curve.
  • asked a question related to Cytology
Question
5 answers
These are the microscopic images of a fruit pulp observed under Binocular microscope at 40x.
Please help me identify the same.
Relevant answer
Answer
I believed that the spring shape is the xylem of the plant
  • asked a question related to Cytology
Question
3 answers
Hello,
We are planning to collect and centrifuge mice bronchoalveolar lavage fluid cells. The downstream analysis we are planning to do (neutrophils, macrophage and lymphocyte % distribution analysis) requires us to transport the cells to another laboratory while maintaining viability. How are the cells normally transported? In what medium, what temperature? How long could the cells be stored?
Thank you!
Relevant answer
Answer
In my opinion, you should not add anything, take the lavage and place in appropriate size tube with minimum PBS just to help transfer lavage to the tube. Transfer the tube at room temperature. Start processing right away at arrival.
I have done with bone arrow (heparinized), upto 4 hours at room temperature, all cells were fine.
  • asked a question related to Cytology
Question
3 answers
I would like to ask whether there will be a worse result of the immunocytochemical reaction, which will be carried out on archival materials (2006-2008): cytological smears of lung cancer, stained by Pappenheim and Papanicolaou?
Relevant answer
Answer
I am afraid it won't work, especially on the previously stained smears.
  • asked a question related to Cytology
Question
9 answers
Dear researchers, according to your practice in field of staining bone marrow cells in rodents.
What are the types of staining and which is the best and easy used differentiating BM cytology.
Dose anyone have atlas or reference guide for rats bone marrow cytology?
Thanks in advance
Dr. Ali Alchalabi
Relevant answer
Answer
If we want to detect infectious agents, there are various histological stains for the agent. however, if we want to look for different cells, we can use immunohistochemical stains. Which different cells do you want to detect?
  • asked a question related to Cytology
  • asked a question related to Cytology
Question
4 answers
Do you think it is possible to study the spread of lung cancer in the parenchyma only by cytological and immunocytochemical methods (if access to histological material is limited)?
Relevant answer
Answer
Kindly check also the following good RG link:
  • asked a question related to Cytology
Question
2 answers
Hello! Help me, please. I have a problem with washing away cells from cytological smears...
Currently, our laboratory uses a manual method of staining cytological smears with immunocytochemical dyes:
1) Fix cytological smears in methyl alcohol for 5 seconds.
2) Stain cytological smears by Pappenheim and Papanicolaou methods.
3) Dissolve 0.01 grams of trypsin in 1 ml of phosphate-buffered saline (PBS) (PH 7.2-7.4) and apply to smears for 10 seconds. (!!!!!) (Are there any errors at this stage? Probably we need to dissolve 0.001 grams of trypsin in 100 microliters PBS? Or reduce the exposure time of the solution from 10 seconds?)
4) Thoroughly wash the smears with PBS.
5) Wash the smears with distilled water to prevent drying.
5) Apply to smears 5% acetic acid solution for 10-15 minutes.
7) Wash the smears again with distilled water.
8) Apply to smears 1% hydrogen peroxide solution for 30 minutes.
9) Thoroughly wash the smears with PBS.
10) Apply the first (І) antibodies to the smears for 1.5 hours (at room temperature or at -4 C per day) in a humid chamber.
11) Wash thoroughly with PBS.
12) Apply the second (ІІ) antibody to the smears: Mix 1 microliter of the second antibody, 10 microliters of human serum 4 (AB) group RH (-) and 100 microliters of PBS and apply to smears for 1.5 hours at room temperature.
13) Thoroughly wash the smears with PBS.
14) Mix 1 milligram of 3 diaminobenzidine tetrachloride (DAB), 6 milliliters of PBS, and 20 microliters of 6% hydrogen peroxide solution and apply to cytological smears for 15 minutes. 15) Thoroughly wash the smears with distilled water.
Please indicate what I am doing wrong, or advise where you can find a better manual technique in which the cells will not be washed away from the cytological smears.
Relevant answer
Answer
Use superfrost plus slides for your smears and take care that they are not too thick. I think the protein digestion is the most critical point in your protocol and the reason why the smears are washed away during the immuno process. If the trypsin pretreatment is necessary I would recommand to prepare a stock solution. Play with different concentrations of the trypsin solution and incubation times. Sometimes a permiabilisation with Triton-x-PBS (0,2% Triton in PBS for 30 min; 3 x 5 min rinsing in PBS afterwards) could be sufficient.
  • asked a question related to Cytology
Question
2 answers
Tell me please, which of p63 antibodies (Manufacturer "Diagnostic biosystems": rabbit RMAB086-01 (antibody titer 1:50); mouse BSB 3605 (antibody titer 1:200) ; mouse BSB3602) is better for staining cytological smears of alveolocyte II type and cells of lung cancer (adenocarcinoma, squamous cell cancer)?
Relevant answer
Answer
Rabbit provides greater benefit over mice when it comes to monoclonal antibodies. Immunoglobulin genes from rabbits form antibodies that fit a much wider range of epitopes than mouse antibodies. Isoforms of a protein that differ by one amino acid residue, results in slight variations in structure. The rabbit's immune system better recognizes these subtle differences, producing monoclonal antibodies.
Moreover, in immunodominance certain epitopes from the same antigen are more immunogenic than others thereby the immune system produces a great amount of antibodies against the dominantly immunogenic epitope, and few antibodies against the other epitopes. In this case, rabbits exhibit less immunodominance than mice.
So, rabbit monoclonal antibody because of the unique features of the rabbit immune system would be more preferred than the mouse monoclonal antibody.
You could also refer to the research article below wherein rabbit monoclonal was preferred over mouse monoclonal antibody for ICC.
So, I recommend that you use p63 rabbit monoclonal antibody for immunocytochemistry in your study because of its higher affinity and increased specificity.
Good Luck.
  • asked a question related to Cytology
Question
3 answers
Hello everyone, I want to know if there is a golden technique in the diagnosis of canine leishmaniasis to evaluate the results of other diagnostic techniques such as ELISA and IFAT and even lymph node or splenic cytology in the latter.
Relevant answer
Answer
urine sedimentation technique
  • asked a question related to Cytology
Question
1 answer
"Approximately 15–30% of all thyroid nodules evaluated with fine-needle aspiration biopsy (FNAB) are classified as cytologically indeterminate. The stepwise unraveling of the molecular etiology of thyroid nodules has provided the basis for a better understanding of indeterminate samples and an opportunity to decrease diagnostic surgery in this group of patients."
Relevant answer
Answer
Recently, TBSRTC, 2nd ed. has been using and Category III, atypia of undetermined significance or follicular lesion of undetermined significance (AUS/FLUS); IV, follicular neoplasm (FN) or suspicious for a follicular neoplasm (SFN); and V, suspicious for malignancy (SM) have been accepted as constituting indeterminate cytology of thyroid nodules. The risk of malignancies (ROMs) for indeterminate cytology are declared as 5-15%, 15-30%, and 60-75% in TBSRTC 1st ed. while 10-30%, 25-40%, and 50-75% in 2nd ed. for Category III, IV, and V, respectively, reflecting a wide range of ROM. The 2016 Royal College of Pathologists, United Kingdom (RCPath, UK) reported indeterminate cytology as Thy 3a, neoplasm possible, atypia/non-diagnostic and Thy 3f, neoplasm possible, suggesting FN, with the ROMs of 20-31% and 24-39%, respectively. In addition, the 2016 RCPath, UK declared Thy 4, SM with the ROM of 70-87%. The 2014 Italian Consensus for the Classification and Reporting of Thyroid Cytology (ICCRTC) divided diagnostic category TIR3, indeterminate cytology, into two subcategories, TIR3A (low-risk indeterminate lesion) and TIR3B (high-risk indeterminate lesion), with different expected ROMs and discrete clinical manners as <10% and 15-30%, respectively. The 2014 ICCRTC also harbors TIR4, SM with an expected ROM of 60-80%. The 2014 Royal College of Pathologists of Australasia (RCPA) and the Australian Society of Cytology (ASC) propounded: i) 3, Indeterminate (AUS/FLUS) with low ROM, 5-13%; ii) 4, Suggestive of an FN (FN/SFN) with moderate ROM, 21-26%; and iii) 5, SM with high ROM, 85-90%. The 2013 Japan Thyroid Association (JTA) Guideline for the Management of Thyroid Nodules declared: i) 3, Indeterminate, 3B, others; Indeterminate, ii) 3A, FNs, 3A-1, favor benign, 3A-2, borderline, 3A-3, favor malignant; iii) 4, Malignancy suspected, respectively. As such, each diagnostic category of the last TBSRTC, 2nd ed., with noninvasive follicular thyroid neoplasm with papillary-like nuclear features, NIFTP, harbor a wide range of implied ROMs.
  • asked a question related to Cytology
Question
3 answers
What is your distinctive clinical approaches for the management of thyroid nodules of both 10-15 mm and over 15 mm, separately, with Category III of indeterminate cytology, TBSRTC, 1st and 2nd ed., harboring high-to-intermediate suspicion sonographic pattern, low clinical risk factors, repeated FNA cytology, molecular testing, or both, are not performed or inconclusive?
Relevant answer
Answer
We might recommend surveillance for the management of the mentioned thyroid nodules, 10-15 mm with Category III of indeterminate cytology, TBSRTC, 1st and 2nd ed., harboring high-to-intermediate suspicion sonographic pattern, low clinical risk factors, repeated FNA cytology, molecular testing, or both, are not performed or inconclusive, comparing the ones >15 mm. This issue deserves further investigation.
  • asked a question related to Cytology
Question
6 answers
I am so curious how ideogram was constructed. I found a publication that used software to construct ideogram, but this software seems like a photoshop software. Is there any proper software to draw or construct an ideogram? thanks ^^
Relevant answer
Answer
For drawing Ideogram you can Visio software as well. It is very easy and applicable. Below is the link if you want to download and use it, but you have to purchase and activate it for the full version. Fortunately, I have the crack and if you are interested I can give it to you.
  • asked a question related to Cytology
Question
3 answers
We are examining tissue from a cluster of human and canine patients with a similar pattern of systemic illness of unknown cause. All members of the cluster have evidence of motile zoospore-like objects in their blood and other tissue aspirates. Control wet-mount preps from healthy relatives of these patients do not show the presence of such motile objects.
Despite the tiny size of these motile objects, the “swimming” motion seems more consistent with the “falling leaf” forward motility pattern associated with a eukaryotic flagella than with bacterial motility patterns. Also, the staining patterns and SEM appearance of these objects appears more consistent with a eukaryote.
Preliminary sequencing studies have suggested sequence homology with stramnopile-type organisms. We are attempting to sequence cultured colonies of the organism but are having extremely low DNA yields despite robust growth of the organism in culture.
We would appreciate the opinion of those familiar with the morphology and zoospore motility of oomycete and related type organisms about the similarities and differences seen in these movies of motility in unstained, aseptically collected adipose tissue nodules from a patient in this cluster, suspected to be infected with a novel or emerging type of eukaryotic pathogen.
Relevant answer
Answer
sure, I'd like to see the pictures - philageis@aol.com
  • asked a question related to Cytology
Question
2 answers
I'm used standard squash technique using fuelgen staining.
Relevant answer
Answer
Nice! Good question....yea..if we know techniques of your experimental, I may tell my students to repeat the same,,, regards!
  • asked a question related to Cytology
Question
3 answers
Hello, Our background is explained in the profile section- but we are a group of veterinarians/other scientists working on an unexpected research project that arose from observations of a cluster of illnesses among dogs and their owners in our veterinary practice. After thorough traditional work-up, we ruled out known causes of this constellation of symptoms and because of the seemingly transmissible nature of the illness, we began looking for perhaps a rare infection. We did find unexplainable objects in urine, cyst fluid, and subcutaneous nodules from the affected animals/humans- and did not see those same objects in blinded control studies using spouses/housemates of the affected individuals. It has taken a while to characterize the shape/properties of these objects since we were seeing them in the context of semi-degraded in tissue or tangled/broken up in urine samples. But now, we have retrieved enough reasonably intact samples to realize that they are large shell-shaped objects, resembling bivalves slightly, that contain long thin complex fibers as structural elements. The hinge/latch-like objects along the length of the fiber appear to bind to other elements in the internal contents of the "shell" and help keep them packaged and organized. I'm quoting "shell" because despite the marked resemblance to some bivalve shells, the shell portion is more dynamic. It can stretch and when the shell opens and the internal contents unfurl, the majority of the tissue forming the "shell" shape actually comes away in plumes of sheets of tissue that unfold in a very organized manner. Intriguingly, these sheets of tissue appear to contain a middle layer with a non-staining fibrous semi-liquid filling that may be mesoglea. The sheets of tissue are made up of connected small versions of almost exactly the same shell body plan as the larger organisms, and these small objects appear connected by a series of tubes that resemble stolons- leading us to wonder if the organism could be some type of hydrozoa or even myxozoan? I've attached a few new photos to demonstrate. We would appreciate any opinions at all about what the identity of these objects may be, and how we can optimize staining protocols +/- DNA extraction techniques to be able to get an identifying sequence for these probable organisms- though there will undoubtedly be some contamination with human/canine DNA.
Relevant answer
Answer
Yes..it seems some invertebrates
  • asked a question related to Cytology
Question
3 answers
Hello, there is more background in our overview page, but briefly- we are a group of veterinarians who came across unusual microscopic findings in a group of dogs and their owners (who happened to also be veterinarians) with an otherwise unexplained syndrome involving hematuria, chronic fibrosis of deep connective tissues, chronic cough, fatigue, and neuropathy. Both the dogs and their owners had a thorough medical work-up in which the cause of illness cold not be determined. Several other close human contacts of the original humans/dogs affected also developed similar symptoms over the following year. Some of the humans have had plateau of their symptoms while others have continued to worsen. The structures shown in attached images were found in the urine, blood, needle aspirates of subcutaneous nodules, and sometimes sputum of the affected individuals (canine and human) and not in healthy family members (matched controls)- this finding was confirmed by blinded readings of the affected vs. control cytology samples. CDC has been informed of these findings and may begin an investigation, though they are hobbled by the current COVID crisis and also relayed that they do not have a pathologist on staff trained to read this type of cytology!? So, we are asking your help in determining if the objects shown do seem to match the criteria for some type of protist (or other organism?), or if they are some type of unusual artifact. We have attempted sequencing without success, but this may be hampered by a thick glue-like mucus that seems to be produced by the organisms binding everything in the near vicinity tightly together. As veterinarians, our knowledge of invertebrate zoology is limited- but collectively, we thought that the objects seemed to resemble cysts of amoebae or perhaps ciliates, or even myxozoans. They are protected by a very stain-resistant outer "shell" (test?) that seems made of aggregates of regional debris. However, some layers stain well with Alcian Blue/Aniline Blue and negative staining with Nigrosan helped to clarify the appearance of some of the objects. The white surface is very birefringent, so we were only able to get clear micrographs when stacking software was used to improve focal range. Some features seemed similar to entamoeba histolytic, but the outer cyst wall is too thick in many examples. Some resembled Blastocystis- with a large central vacuole, but again, there were inconsistencies with that identification as well. If anyone can point out specific details/features that suggest real organism vs artifact, and/or label any of the details for us to give us some landmarks to follow (we though we were seeing distinct nuclei, but were not certain...) that would be extremely helpful.
Relevant answer
Answer
Hi
Photos show artifacts and no structures seen at all related to a protozoan, all protozoa usually have regular structures in various forms. Search the slide again carefully and also with high magnification!
good luck
  • asked a question related to Cytology
Question
5 answers
Hello everyone,
I am looking for a scientist who have experance in histopathology and cytology to advise me the essential equipments that using in Histopathology and cytology Laboratory. We want to establish this Laboratory in our new centre for cancer treatment.
THANK YOU
Relevant answer
Answer
Good day,
Equipment of Histopathology and Cytology lab depends from volume of biopsies which is planning to be, the clinical departments etc. I can give you some advises about Histopathology equipment, in my country Histopathology and Cytology are different departments.
Best regards,
Dmitry
  • asked a question related to Cytology
Question
10 answers
In a patient with multinodular goitre ( non toxic and no compressive symptoms) , with a single suspicious nodule (2 cm) on ultrasonography and cytology of that nodule as Bethesda V ( suspicious of papillary carcinoma), what would the extent of initial surgery be? Hemithyroidectomy? Total thyroidectomy?
Relevant answer
Answer
If the contralateral lobe is normal and is not an unfavorable histological variety, for a lesion of that size both procedures are acceptable
  • asked a question related to Cytology
Question
3 answers
Full history is in the project description, but briefly- a team of other veterinarians/colleagues and myself have stumbled across a case of multiple animals that have similar symptoms and are co-housed. Their presentation suggests an infectious disease, but no known infectious agent could be found despite thorough work-up. However, cytology samples from clean aspirates of subcutaneous nodules and urine filtrate (they have bloody urine) seems to be showing repeated structures that do not appear mammalian in origin. We seek to understand if these structures are some type of very organized artifact, or if they instead suggest that there may be some very unusual/novel type of organism (Annelid or Polychaete esp.) causing/involved-in these animals illnesses. Thank you for your time!
Relevant answer
Answer
@ Matthew B Paddock; Thanks for your reply. We did attempt 16s sequencing of a tissue sample taken from one of the subcutaneous nodules (before we realized that there could be a eukaryote involved). The 16S sequencing (from RNA extracted from tissue blocks from what should have been a surgically sterile site) did show a mixture of many bacterial species. However, the profile wasn't anything that would suggest a relative of a known pathogen, nor did it suggest surface contamination (ie- no staph species etc..) The profile didn't have any one main dominant species either, but was 20% this, 18% that, 15% this etc.... I don't have the profile in front of me right now, but, from memory, the bacterial species in the tissue sample were unusual, with an overrepresentation of extremophiles. At first I interpreted this as potentially some species that might be surviving our autoclaving of instruments (ie- could tolerate the autoclave temperatures but not typically mammalian pathogens so were not the cause of disease in the animals). But, once the observation of the filaments/worm-like organisms was made, it did occur to me to that perhaps if the infection were eukaryotic, perhaps the profile of bacteria seen could reflect the composition of the microbiome of the putative novel parasite. I cross-referenced the profile of bacteria with the known microbiome profile of c-elegans and, though not an exact match, the proportions and types of bacterial species represented did suggest that perhaps the species identified could represent the bacterial population of some eukaryotic species. Our first thought was that the putative novel species was some type of nematode/filarial infection. We did attempt to have the tissue sample probed with nematode-specific 18S rRNA primers to see if we could amplify/sequence any nematode DNA. Interestingly, the primers did bind and amplify a target sequence, but when that was attempted to be sequenced, the PCR products were not clean enough to yield an identifiable species. Later, other microscopic findings led us to consider other phyla (beyond nematodes/trematodes). Since this is mostly a project done out of curiosity rather than our primary line of work, we have not had the budget to proceed with any additional PCR studies. If you would like to see the actual genus/species break-down for the 16S analysis, I'd be happy to dig it up and post it for you (just don't have access to that computer today).
  • asked a question related to Cytology
Question
8 answers
Hi. As far as I understand you just be having histology as your gold standard. What I want to understand is what were your true negatives ? Say if p16 and ki 67 came out negative on cytology how did you confirm it's a true negative? Did you have another negative group?or you took the same histology as negative control too?
Relevant answer
Answer
I am reading your comment here now.
Yes, Nikhat. Both Tauangtham and I have understood your question the first time around.
It seems though that you did not read carefully the scientific links that I have sent you.
If you had, you would have understood that it is long established that there is no such thing as a 100% fool-proof diagnosis of benign, pre-cancerous or cancerous histology, based on three imperfect tests, the scope of which (esp. sensitivity & specificity) we still explore, and that are, therefore combined to complement each other.
In this format (and without knowing the life style, the medical history or the current physiology of the women that are being tested), these three tests used once will never give a 100% negative testing results. Your supervisor is correct.
You are testing living creatures with complex anatomy and physiology, whose body condition changes dynamically. Anomalous cytology is present at all times, but some of it is successfully cleared by the immune system, while other is not.
It takes time for persistent anomaly to develop to a degree sufficient to diagnose in histology.
Testing negative for diffuse p16 today does not automatically mean health, or that the same woman is going to test negative for diffuse p16 again in three-months' time:
Diagnosis would also depend on lab guidelines, as well as patho-histologist's personal judgement and experience.
The last link I have sent you is a dissertation that discusses new DNA methylation markers to try and better the success rate of the three testing tools you work with.
As a physician, I will argue that (and I have already told you via messaging) in the face of:
-- a pronounced lack of certainty on how many diseases develop, and
- imperfect test data on patients' condition,
it is unethical (to recommend to patients or) not to argue in favor of exploring all avenues (even invasive ones) to:
- make an informed diagnosis, (and by doing so)
- save patients more anxiety, suffering and stronger medication (with negative side-effects) due to delayed/incorrectly made diagnosis, and
- ensure that patients stay alive (by detecting pre-cancerous and cancerous conditions early)
I hope this brings further clarification.
  • asked a question related to Cytology
Question
4 answers
Morphological, cytological, biochemical and molecular tools are used to differentiate Two species. Which is the best and accurate method?
Relevant answer
Answer
I agree with Valeria Tananska , the reasoning being that morphological differentiation has been utilized exclusively, until recently. However, these days, molecular methods can provide exact quantification of the relatedness between two organisms. The issue here is that the molecular methods are not as well developed and as common to practice as morphology. As molecular methods become more commonplace within organism differentiation we will see a different, and hopefully more exact, classification of species. This does also mean that different classification methods might showcase alternative features, and perhaps even the opposite of what you would expect, so the methods might diverge in interpretation.
  • asked a question related to Cytology
Question
3 answers
Is it possible to extract DNA for further analysis/diagnosis from Papanicolaou stained cytology slide?
Relevant answer
Answer
Zohar gavish the Answer of Helena is correct
  • asked a question related to Cytology
Question
3 answers
Noninvasive follicular thyroid neoplasm with papillary like nuclear features is a very good topic for discussion.
Relevant answer
Answer
You can work with cytological specimens both retro- and prospective but (see the answer of Olena Polyakova) will need histological (surgical) material to make the diagnosis of NIFTP.
  • asked a question related to Cytology
Question
2 answers
Hello!
After daily vaginal lavages, we have had a mouse who appears to have been in estrus for three days in a row based off of cytology and the presence of only cornfield epithelial cells. The samples typically dry clear, but as you can see in the photo this one dried very white. Also, yesterday both of her samples washed off the slide after ample drying time and our standard 1% toluidine blue staining protocol.
Any idea what might be happening to this mouse in terms of her cycle?
Thank you in advance!
Relevant answer
Answer
it must be diluted or spread in order to be visualized well under microscope because it looks thick
  • asked a question related to Cytology
Question
21 answers
Many a times we don't get good cellularity to give a clear diagnosis of ovary in cytology. Any advise please?
Relevant answer
Answer
I still feel as and when Ovarian lesions are excised, one can prepare a set of good well fixed slides for study and learning from classical lesions.
  • asked a question related to Cytology
Question
2 answers
In literature there are different methods to detect hpv on bladder tumour studies. Frozen sections, direct tumour tissue, urine, cytology. Using PCR assay I'm planning a study on bladder tumour & hpv. Which sampling method do you recommend?
Relevant answer
Answer
actually I wanted to study on fresh samples. So urethral swab and first void samples I collected.
  • asked a question related to Cytology
Question
5 answers
Dear Sir/ Ma'am
Can any researcher share/provide the reprint of following articles (Avdulov 1928, 1931) on grass cyto-systematics...
1. Avdulov, N. P. (1928). Systematicheskaya kariologiya semeestva Gramineae. Drievnik vsesojuznogo Sezda Bot. Leningrade 1928: 65-66.
2. Avdulov, N. P. (1931). Karyo-systematische Untersuchungen der Familie Gramineen. Bull. Appl. Bot., Genet. & Plant Breeding Suppl. 43: 1-438.
Relevant answer
Answer
I can only provide you with Avdulov (1931) in Russian version (>270 pp), German version is not available.
The file is attached. You will need DJVU player software in order to open the file (free download available from https://www.cuminas.jp/en/downloads/download/?pid=1 )
  • asked a question related to Cytology
Question
5 answers
The taxonomy of the genus Crocus is extremely complicated due to the lack of clear distinctive characters, the wide range of habitats and the heterogeneity of the morphological traits and cytological data.
Moreover they show that there are several problems at the infraspecific level. Genetic dissimilarities between the known subspecies revealed that most of the subspecies must be ranked as species.
Relevant answer
Answer
The genus Crocus L. belongs to the large family Iridaceae and is a systematically problematic genus. In the Old World, about 100 species are known (Harpke et al., 2013). Although molecular studies have been increasingly used to examine the phylogeny of living organisms, they have only recently been applied to the genus Crocus (Petersen et al., 2008). DNA markers have been used to characterize germplasm collections, inform breeding programs, and facilitate genetic diversity studies and taxonomic analysis. The utilized methods include interretrotransposon amplified polymorphism (IRAP) (AlaviKia et al., 2008), random amplified polymorphic DNA (RAPD) (Grilli Caiola et al., 2004; Beiki et al., 2010; Rubio-Moraga et al., 2010), amplified fragment length polymorphism (AFLP) (Zubor et al., 2004; Erol et al., 2011; Nazzal et al., 2011), intersimple sequence repeat (ISSR) (Rubio-Moraga et al., 2010), and simple sequence repeat (SSR) (Rubio-Moraga et al., 2010; Nemati et al., 2012) analyses. Among these molecular markers, AFLPs were found to show high levels of polymorphism per primer pair and yield high resolution and reproducibility (Meudt and Clarke, 2007). Fore more information follow this link:
  • asked a question related to Cytology
Question
5 answers
Currently, I would like to study more about seed development and apomixis. I want to check the ploidy of endosperm of seed in the case apomixis happens. I know one way is to determine by using flow cytometry. But my lab is not keen on cytology. I wonder if there is another simple way or not? Thank you so much for reading my question.
Relevant answer
Answer
Dear Dung
In the attached papers, apomixis frequency and ploidy levels in Poa and Centotheca using flow cytometric techniques were examined, which I hope will be of interest to you.
  • asked a question related to Cytology
Question
3 answers
In this regards, I wish to point out that there is gross miss concept regarding soul & rebirth among almost all ethnic groups of human societies. I wish that people should rethink the concept of Rebirth & Germ Cell Soul & (not sole) & based on Science of Genetics-classical, cytological & molecular) the point of thinking is just similar as matter is not destroyed but transform into solid gas & liquid in the way germ cell egg & sperm) are not destroyed but continue the life through individuals female & male generation after generation. this phenomenon I call it "THE BIRTH''
Relevant answer
Answer
Kindly elaborate about the essence of your project. Are your findings against the soul theory mentioned in Geeta or Kriya Yoga techniqs....
  • asked a question related to Cytology
Question
4 answers
Protandim and NRF2 stimulator do that. Any researcher want to add some thing, I cordially invite them. I am very thankful to your suggestion.
Relevant answer
  • asked a question related to Cytology
Question
1 answer
Does anyone have a recipe for Saccomanno fixative (a cytology fixative) for sputum preservation?
Relevant answer
Answer
Hello Hamdia,
Saccomanno's Fixative is an excellent pre-fixative for cytology specimens and the fixative of choice for collecting and fixing sputum specimens for cytology. This fixative in in fact, one of the most widely used fixatives in cytology that is recommended for cytology specimens such as sputums, urine, FNA'S, bronchial washings, pleural and peritoneal fluids, and ThinPrep™ preparations. The formula is accessable at the website indicated below:
Saccomanno's fixative is 50% ethyl alcohol which contains approximately 2% of Carbowax 1540 (Union Carbide Corporation, UCAR). Carbowax 1540 is solid at room temperature, with a melting point of 43 to 46 C. To avoid having to melt it whenever the fixative is prepared, a stock solution can be prepared by melting Carbowax (melted in an incubator or hot air oven at 50 to 100 C) and adding it to an equal volume of water or 50% ethyl alcohol. The mixture will not solidify. Saccomanno's fixative can then be prepared with 430 mL of water, 530 mL of 95% ethanol, and 40 mL of the stock Carbowax solution. Some light green SF or fast green FCF can be added to color the fixative. Koss warns that the denaturants in reagent alcohol may cause excessive hardening of mucus.
ref: Histology FAQ
Staining, Histochemistry and Histotechnology
(Frequently Asked Questions)
Dr. John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada
With best regards,
ROWEN T. YOLO, M.D.
Dept of Pathology
Faculty of Medicine and Surgery
University of Santo Tomas
Manila,Philippines
  • asked a question related to Cytology
Question
22 answers
My group is working on embryo rescue in Cucumis genus. We cultured the immature seeds and we got a lot of types of germination. Some seeds germinated as usual. But almost seeds regenerated callus. And I think we got two kinds of calli (embryogenic callus and non-embryogenic callus). The problem is that my lab is not expert in cytology or tissue culture. So that, I want to identify them in the simple way but correction. I am looking forward to your suggestions. Thank you in advances!
Relevant answer
Answer
Under the microscope the sotic emryo cells are round in shape. Highly cytoplasmic densely stained more over the starch grains are accumulated in the cytoplasma. See the attached file please
  • asked a question related to Cytology
Question
6 answers
the patient is 18 years old complains of weight loss, abdominal distention and menstrual irregularities. Is there a role for FNAC?
Relevant answer
Answer
Hello Dr Noha Hassan
I fully agree with the opinion of the others wherein correlation, with clinical presentation, biochemistry (AFP-B-HCG), and ultrasonographic findings point out that this is most likely a germ cell tumour of the ovary. In an 18 year old woman, this is most likely a mature cystic teratoma involving both ovaries which is not uncommon. The findings of ascitis and omentoperitoneal thickening may perhaps suggest peritoneal irritation with, in my opinion, mesothelial reactive changes that is secondary to the compression effects of the huge ovarian masses, however of course, I cannot discount the possibility of a malignant teratoma with peritoneal metastasis, albeit this is rare in this age group. Exploratory laparotomy with ovarian resection (or frozen section), peritoneal implant biopsy, and ascitic fluid cytology, would be the management of choice. In my opinion a pre-operative ultrasound-guided percutaneous FNAB may yield not much of a helpful information as far as diagnostic determination between a benign vs malignant teratoma is concerned. A sample of the ascitic fluid of course may be collected for cytodiagnosis, but may well prove to be of limited value in this case. I would be most interested to follow up this case. Please update me with any latest findings. Thank you very much for sharing this interesting case.
  • asked a question related to Cytology
Question
2 answers
Looking for ways to improve cell block for cytology.
Relevant answer
Answer
Hello, in my lab we use HistoGel - Thermo Scientific (Ref. HG-4000-012). The first spin (sample +fixative are made at 2800rpm (aprox. 1300-1500g) for 5 min. The second spin (sample + histogel)  3500rpm (2000-2250g) for 1 min.
Hope it helps
  • asked a question related to Cytology
Question
4 answers
I don't know how to adjust bonferroni correction to my data. 
Firstly, I tested the levels of several types of biomarker candidates in normal and cancer serum using ELISA. I found three types of potential biomarkers (A, B, C) for discriminating cervical cancer from normal cytology. 
Secondly, I combined three types of biomarkers using logistic regression model. 
Then, I wanna using bonferroni correction to reduce type I error but I have no idea how to adjust the bonferroni correction to my data. 
Is the bonferroni corrected p value < 0.016? (<0.05/3 biomarkers)
Thank you for anyone's help. 
Relevant answer
Answer
Yes, for Bonferroni correction you did correct. Basically, here are 2 ways of doing it and both lead to the same result: one way, as you did: when deviding threshold level (0.05) by number of tests. And the second way: to multiply obtained p-values for each test by the number of tests and if the adjusted p-value is still below 0.05, consider as significant. This is about Bonferroni correction.
But now, before doing the Bonferroni correction, clarify for yourself: did you do multiple logistic regression of univariate? If you did multiple logistic regression, you do not need any further correction. As I understood, you combined them, meaning that you did multivariate logistic regressoin analysis. So, just use the p-value you obtained. You could apply the Bonferroni correction to the initial analysis, when all biomarker were analysed between normal and cancer serum.
  • asked a question related to Cytology
Question
2 answers
Hi there,
we are focused on impression cytology of conjunctiva. We would like to isolate the cells from imprints on strips of Millipore filter membranes. We´ve tried mechanical isolation/detachement into the DMEM with microcentrifuge tube shaker and then vortexing. However the yield and cell condition are not good. Simply, the most of the cells are damaged. Does anyone have some experience with the cell isolation from filter membranes? Any hint can be helpful. Thanks a lot!
Relevant answer
Answer
In the publication attached, they use laser microdissection (MMI CellCut) to isolate cells from ISET filters. I hope this helps.
  • asked a question related to Cytology
Question
2 answers
I am studying siphonophore tentacle batteries across many species, using different microscopy techniques. I think I might have found the largest nematocysts known for all Cnidaria, as far as my literature search goes, but maybe someone has found something larger. Also, let me know if you heard of any intracellular structure larger than nematocysts.
Relevant answer
Answer
Thanks Joan! The ones I've measured are way larger than those, heteronemes >270um capsule length, >1M um3. I don't have discharge tubule length values for them yet, but it would be interesting to record that too.
  • asked a question related to Cytology
Question
1 answer
I've noticed several contributors on here implicating that ATP synthase could operate outside on the mitochondria. Under Peter Mitchels chemosomotic theory it is possible that the membrane potential created by calcium ion concentrations in endoplasmic reticulums could drive hydrogen ions down ATP synthase if present in the ERs membrane. I've spent sometime thinking how this could be demonstrated, unfortunately I have fallen short as the majority of assays i'm aware of use false substrates for the citric acid cycle to demonstrate ATP synthesis..
Relevant answer
Answer
Sorry , Idont no
  • asked a question related to Cytology
Question
2 answers
I would like to perform a immunocytochemistry (ICC) on differentiated THP-1 to evaluate the expression of a membrane receptor. I will use for that a primary conjugated antibody. On blocking steps do I really need to use a Fc receptor blocking solution or can I use regular blocking agents (like FBS or BSA)?
Relevant answer
Answer
In our experience we have saw binding of conjugated antibodies to macrophages only once. So technically it could be possible. Since serum contains immunoglobulins (in contrast to BSA) thus using of BSA or pooled human heat inactivated AB serum could be used as source of a concurrent ligands for Fc receptor and effectively downgrades false positive signal (if exist) to minimum.
  • asked a question related to Cytology
Question
2 answers
1. To germinate the seeds on petriplate and use the meristems of the root and dip into the nanoparticles solution for 0,1, 2 hrs ?
Or
2.First dip the seeds in nanoparticles solution for desired period and then allow it to germinate and then fix the root meristems for cytology assay?
Relevant answer
Answer
Dear Melissa Chernick,
Thanks for adding the answer along with reference papers.
  • asked a question related to Cytology
Question
1 answer
Does anyone have experience with Merk's Neoclear and Neomount solutions? Does this require additional steps after the standard Papanicolaou staining procedure?
Relevant answer
Answer
Tibor, just asking about your standard PAP-staining: which one do you use (when reading:
with 3 procedures: 
i)    Procedure 1 (Standard Method)
ii)   Procedure 2 (Modified Pap Procedure)    and 
iii)  Procedure 3 (Rapid Economic, Acetic Acid, Papanicolaou Stain Method)
I am wondering why you did not get any answer since 2012! (but admit that I found your question some minute ago....)....would recommend, to edit your question and add some keywords more: e.g.: Histology, Histochemistry, smears, Staining   to eventually increase readership.... best wishes and good luck...W.M.
NB: I haven't done PAP-staining during my former professional career...but perhaps I could help -at least knowing the protocol of your PAP-stain....
  • asked a question related to Cytology
Question
4 answers
I am veterinary pathologist and I am working on Pneumocystis in pigs. We want to establish a method and would need unstained slides from Pneumocystis jirovecii positive AND negative humans. Most publications deal with PCR or cytology, therefore it is really difficult to find somebody who may have formalin fixed paraffin embedded material. I really hope that there is any possibility to get material by this way. Thank you!
Relevant answer
Dear Ivan,
we will use the native samples. A method has been described for human Pneumocystis and we will try to adapt the protocol.
Greatings back from my colleagues!
Christiane
  • asked a question related to Cytology
Question
9 answers
Hi all,
I would have to irradiate lymphoma cell lines (Bl41 and J3D) and planning on looking for DNA damage clearance by IF following different nuclear markers. The center where I will irradiate the cells has no cytospin and so for short time point I would not be able to get the cells to my lab on time.
One solution would be to fix the cells before cytospin but I cannot find any protocol for that. Has anybody try to fix floating cells before cytospin?  
Thanks,
Matt
Relevant answer
Answer
No issue at all, I developed techniques over 20 years ago. Simply re-suspend your cells in medium and add formaldehyde (1:10 producing a 4% solution of formalin). I recommend using 'charged' slides for your cytospins and I spin at 500rpm for 10 minutes. The remaining cell suspension can quite happily be stored in the fridge for 12 months+ and more cytospins made at any point. I've used this for HCC, breast cancer, prostate cancer cell lines to name but a few. I've attached images of EBV+ lymphoma cell lines to demonstrate the results.
Formalin fixation obviously will then require some form of antigen retrieval technique, pre-immunostaining. Having been at the forefront in developing these in the UK in the early 90's I actually published on the use of microwave antigen retrieval for cytological preparations (Reynolds et al Cytopathology 1994; 5: 345-358).
Some say this is problematic for fluorescent techniques, but that is a myth. I use pretty much the same protocols for light or fluorescent microscopy, particularly on formalin fixed paraffin embedded tissues, as evident by my numerous publications showing this, eg Reynolds et al Hepatology. 2008;47(2):418-27.
Hope this resolves your problem and please feel free to contact me direct if you experience any specific issues.
Regards
Gary
 
  • asked a question related to Cytology
Question
6 answers
Hi all,
Here I attach the snapshot of the imaging area. These are RBC's forming clusters which affects my further calculations/experiment. How do I avoid this? Any coating to the channel would help? Thanks in advance! 
Relevant answer
Answer
Try to wash flow-chamber  (before measurements) by buffer that supplemented with 0.5-1.0 % of albumin.
  • asked a question related to Cytology
Question
3 answers
Cytologist, wheat, mitosis   
Relevant answer
Answer
Hi, Nader,
Cytogenetic plant (wheat) techniques, namely, Fixation, Pretreatment, and Staining, please, see at site
Godspeed
  • asked a question related to Cytology
Question
4 answers
I have always worked with whole tissues. Now I will work with cells. Granulosa cells will be obtained form follicular fluid (from follicle aspiration) after centrifugation. As I have never worked with these samples, I´m looking for a protocol to collect them in a slide, fix them, preserve them for a month and then perform an immunocytochemistry. thanks!
Relevant answer
Answer
You can make a cell block the technique you can found everywhere. 
But in my experience, instead of using commercial gel (ex Histogel), we use Agar gel 3%, it works well. 
  • asked a question related to Cytology
Question
3 answers
Hi,
I'm trying to analyze the ecological variation in two cytotypes of the same plant species. I do not have environmental data collected myself. I would like to use open source data such as "worldClime". I would like to see if there is a difference in the distribution of the two cytotypes in terms of environmental factors. I use R for my analysis. I would much appreciate suggestions about appropriate methods.
Thanks
Relevant answer
Answer
Dear Piyal
Though I never work on the ecological variation analysis methodology, but as a researcher on plant taxonomy I can guess how tough your work is.  actually for any sort of ecological variation analysis or study your own data about the climatic condition of the study area is needed, other wise if you depends on the data available in the internet system may distract your experiment due those data are not accurate in-every sense.  
besides these during analysis of the cytological variation it is also very important to take a grave look on the environmental condition because the expression of the gen often depends on various environmental condition. 
with best wishes
Sunit Mitra 
  • asked a question related to Cytology
Question
9 answers
Attached I send two documents in PDF with photographs of the sample, stained with MGG, another with Pap. Final magnification of 400x.
Relevant answer
Answer
these cells are the size of lymphocytes with very dense nuclei, I think  they could be apoptotic cells, perhaps due to therapy?
you can try confirm apoptotic cells by immunocytochemical detection of c-caspase 3?
  • asked a question related to Cytology
Question
8 answers
I use rat ant-mouse CD8a Ab(from eBioscience) to stain mouse pancreas and the Ab always stains the cells of secretory acini which looks like "specific binding", the "specific binding" not in islet. The CD8 antibody works well for mouse spleen,thymus, lymph node. I think it's pancreas tissue problem,  no matter how I try , like bake slides,  deparaffin, antigen retrieval, block, Ab incubation time and temperature, wash….the secretory acini staining always there, some area strong, some weak, no particular pattern(I'm sure it's not CD8 antigen). How can I get rid of the "specific" non specific binding? Thanks.
Relevant answer
Answer
Hi Maria;
Thank you very much. Your answer is very helpful and I'm going to use the 2 reagents for CD8 staining. So far for pancreas IHC , only CD8 has the problem, I did other IHC with primary Abs from rat and from rabbit, no such problem.
  • asked a question related to Cytology
Question
15 answers
Can anybody help me to get the protocol for wright staining? I want to stain the different cell present in sputum. 
Relevant answer
Answer
To Hamid:
despite the request he posted long time ago I would like to add here for the convenience of others:
go to the Reply from  Tam Quyet Nguyen (this thread, # 08) and you'll find the Sigma-Aldrich protocol....
best wishes and regards, Wolfgang
  • asked a question related to Cytology
Question
1 answer
I am studying 'chromosomal count' and 'mitosis' in tree plant. I have germinated the seeds in petri plate and want to study allelopathic effect of plant extract on cell division. Kindly help me in root tip treatment and staining technique for better results.
Relevant answer
Answer
sorry I am not a botany is not my speciality K.
  • asked a question related to Cytology
Question
5 answers
I am interested in acquiring a Doncaster counting dish to count AM fungal spores since the use of the Burker-Turk or Neubauer chamber is not suitable for this. Many spores are bigger than 100 µm so they get stucked between the two glasses. Counting directly in a petri dish is not easy since you don't have a path to follow and is easy to get disoriented. Thus, the Doncaster chamber seems to be the best option but I cannot find where to buy it. Any advice about providers or alternatives? Thanks in advance.
Relevant answer
Answer
Nosotros las conseguimos con colegas venezolanos del Instituto Venezolano de Investigaciones Científicas (IVIC), por favor explora estos contactos: Dra. Milagros Lovera: mlovera@ivic.gob.ve y Dra. Alicia Cáceres:alicia2001@gmail.com. Suerte con esta gestión, saludos cordiales de Eduardo
  • asked a question related to Cytology
Question
4 answers
In 1869, the resident physician at Melbourne hospital, Australia, Thomas Ashworth for the first time observed cells from a postmortem blood sample, from a patient who had about 30 subcutaneous tumors, that were morphologically identical with those of the cancer itself. This observation prompted him to suggest they “may tend to throw some light upon the mode of origin of multiple tumours existing in the same person”.
Relevant answer
Answer
You don't happen to have the publication available somewhere ... and a photo of Thomas Ashworth?
  • asked a question related to Cytology
Question
1 answer
I am looking for some databases to apply some techniques for the detection and classification of abnormalities in images of cytological tests.
Relevant answer
Answer
I am dealing with invertebrate tissues but my supervisor ifor bowen has a lot of papers about cancers you can search for i.d. bowen and also cathrin saraf
  • asked a question related to Cytology
Question
1 answer
Has anyone ever made their own PTFE-coated microscope slides? The ready-made PTFE-coated microscope slides are sold at a high price in my country, so I am wondering if it is more cost-effective if I simply buy a PTFE tape, and stick it to the available glass slides (we have drawers of glass slides in stock at the moment). What do you think?
Relevant answer
Answer
You could use a silane such as 1H,1H,2H,2H-perfluorodecyltrichlorosilane, commonly abbreviated as PFDTS.
It is rather inexpensive and I think it would do the job.
  • asked a question related to Cytology
Question
2 answers
Hey!
I just started working with NIH3T3 Cells and this week I conducted my second plasmid DNA transfection with lipofectamine2000 in this cell line.
Unlike the first time, which worked perfectly, 3 hours into the transfection I noticed my cells started dying and the survivors nuclei started looking like crescents or really thin donuts or onion rings and more like epithelial cells rather than fibroblasts. The transfection didn't work.
I work a lot with SN4741 cells and I used my SN4741 cell media to culture the 3T3 cells as all the others in my group are doing the same with no deleterious effects on the cells.
The main difference between our SN media and the 3T3 media recommended by ATCC is the serum (we use FBS and not BCS) and we supplement the media with extra L-Glutamine and Glucose.
Can someone tell me if this morphology is to be expected or if it is otherwise abnormal?
Relevant answer
Answer
Hi,
Maybe you should also take care of the quality/purity of your plasmid DNA, e. g.  by using an endotoxin-free plasmid kit when preparation from bacteria. The amount of DNA might also be a critical factor. Also, try out transfection without serum, if you did not so. Good luck.
  • asked a question related to Cytology
Question
2 answers
Hello everyone,
Is there any solution to reduce considerably the 4T1 mammospheres aggregation.Is there any other agent that can replace methyl cellulose? 
Relevant answer
Answer
Please see these papers. They may help you out.
Cancer Letters vol.323(1) 2012 20-28.
Good Luck!
  • asked a question related to Cytology
Question
4 answers
This is a picture about IHC result. We found that the protein was highly expressed in the nuclear and cytoplasm of spermatocyte, round  spermatids, spematogonia and Sertoli cells,  but was  hardly detectable in elongated  spermatozoon. I am confuesed how to distinguish second spermatocytes and round spermatids.
Relevant answer
Answer
I agree with Laura and Shahzad, it is not easy to detect secondary spermatocytes, specially when tubular sections are not completely obliques (since maturation stages are mixed). Performing complementary histological sections with PAS staining  could help you to differentiate stage XII, and thus secondary spermatocytes. But tubules must be completely rounded and with the lumen visible. You can look up my last work in which I combine immunohistological techniches with PAS staining in mouse testis.
  • asked a question related to Cytology
Question
5 answers
What are the differences between cytology, cytogenetics, and karyology?
How do they deal with each other?
Relevant answer
Answer
Agreed with Andrew - apart from the fact that cytogenetics covers whole field of chromosome research - not only in human, but also animal, plant, fungi...
Also male human karyotype reads as 46,XY according to ISCN(2013).
As soon as you do molecular cytogenetics in interphase nuclei you also may say here you start to enter the field karyology.
  • asked a question related to Cytology
Question
6 answers
Hello
I have a really confusing question. I wanna count the cell of liquid based cytology(LBC). But the cell morphology is different between normal LBC and cancer LBC. And normally, cancer LBC contains so many sediments which with red color. I have no idea that if the red sediments also are cells or may be other debris. 
Could anyone help me?
Thank you!
Relevant answer
Answer
Cell counting depends on the type of cells you require.If the sample contain only one type of cells,you can use an image analysis software easily.Otherwise you have to train the software first.
  • asked a question related to Cytology
Question
4 answers
I have read Levan et al. 1964 and Atlas of Mammalian Chromosomes, but I am still want to know proper nomenclatur of mammalian chromosome pair number, especially in bats and rats. Thank you so much
Relevant answer
Answer
Thank you so much Prof. DC Gautam
  • asked a question related to Cytology
Question
5 answers
Cervical cytology images contain nuclei,cytoplasm and background. If the nuclei overlap each other what method can be used for segmentation
Relevant answer
Answer
There are different methods for touching nuclei seperation in literature. Before segmentation find out the seed points accurately. Euclidian distance map with multy scale laplacian of gaussian, radial symmetry transform,single path voting with shifted gaussian and concavity point detection are some of the technique seen in the literature. I also need matlab code for these. if any one has the code,pls help  
  • asked a question related to Cytology
Question
6 answers
I found a publication about chromatid break in mice. The mice were induced by some chemical subtances. So, in the natural condition (wild) what's factor that makes chromatid break happen?. Thank you ^^
Relevant answer
Answer
There are several factors causing chromatid breaks in natural conditions: i) replication across a nick would give rise to chromatid breaks; ii) a second cause is oxidation from radicals occurring during normal oxidative respiration; iii) failures of DNA  topoisomerases; and  iv) natural ionizing radiation including gamma rays and X-rays.
  • asked a question related to Cytology
Question
5 answers
I have conducted karyotype of bone marrow in the wild
But, I have never try karyotype using blood. I need some methods that really useful and work. I think blood is more easily contaminated, so it must quite difficult
Thanks
Relevant answer
Answer
Sounds good thank you so much for both of you Sonal and Khaled :D
I have to try it
  • asked a question related to Cytology
Question
2 answers
Dear Researchers, I have been having difficulties in determining the chromosome numbers of some accessions of Hibiscus sabdariffa germplasm in Nigeria. Common cytological procedures for both mitosis and meiosis had ben used, yet no result. What should I do?
Relevant answer
Answer
@Please enlist the difficulties/observations you have come across.
Regards
  • asked a question related to Cytology
Question
6 answers
When red blood cells are stimulated by toxic compounds (e.g. mercury) they change morphology to acanthocytes. Most part of papers report SEM analysis; is it possible to apply a flow cytometric method?
Relevant answer
In theory, you might be able to identify your acanthocyte population simply by using Forward and Side scatter, as these cells appear to be smaller and possibly more granular than typical erythrocytes. Have you ever looked at your control and toxin treated cells via SSC/FSC? Happy to have a look, if you could post a picture of your flow cytometry data.
AnnexinV is used to detect phophatidylserine exposure - this is for example the case for apoptotic neutrophils that can be thus phagocytosed by macrophages. I have found one article, where AnnexinV was used to detect changes in the phospholipid structure of erythrocyte membranes (PMID: 8562945), so might indeed be worth using this as a stain.
  • asked a question related to Cytology
Question
4 answers
cytological criterias are helpful ?
Relevant answer
Answer
The following article gives answer to the question
Sarcoma. Jun 2003; 7(2): 87–91.
doi: 10.1080/1357714031000081207PMCID: PMC2395518Malignant Glomus Tumour: A Case Report and Review of the Literature
Annarosaria De Chiara,1 Gaetano Apice,2 Stefano Mori,3 Giustino Silvestro,4 Simona N. Losito,1 Gerardo Botti,1 and Vito Ninfo5
1 Department of Pathology, Istituto dei Tumori di Napoli ‘G. Pascale’ di Napoli, Via M. Semmola, Napoli, 80131, Italy, 2 Division of Medical Oncology B, Istituto dei Tumori di Napoli ‘G. Pascale’ di Napoli, Napoli, Italy, 3 Division of Surgical Oncology B, Istituto dei Tumori di Napoli ‘G. Pascale’ di Napoli, Napoli, Italy, 4 Division of Radiotherapy, Istituto dei Tumori di Napoli ‘G. Pascale’ di Napoli, Napoli, Italy, 5 Department of Pathology, Facoltà di Medicina e Chirurgia di Padova, Padova, Italy, Annarosaria De Chiara, Email: moc.liamtoh@araihcedanna.Corresponding author.
Author information ► Copyright and License information ►
Copyright © 2003 Hindawi Publishing Corporation.This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
This article has been cited by other articles in PMC.
Abstract
Purpose: Glomus tumours are characteristically benign solitary tumours. At our knowledge, about 23 reports are present in literature regarding the malignant counterpart, but only a minority developed metastases. We describe a locally aggressive glomus tumour with lymphnode metastasis.
Patient: The patient was a 40 year-old man presenting a 1.5-cm lesion on the right wrist incompletely excised and a recurrent tumour, 4 × 2 cm in size, removed after 9 months, for which he received radiotherapy. After 2 years he developed an axillary lymphnode metastasis.
Results: Histologically, both tumours (primary and metastasis) were similar. There were sheets and nests of uniform small cells with scant eosinophilic cytoplasm and round to polygonal nuclei; there was some degree of pleomorphism and the mitotic index was high (up to 18 m/10 HPF). The tumour cells were positive for vimentin and smooth muscle actin, but negative for desmin, NSE, Factor VIII, chromogranin, cytokeratin. Remarkably, in the primary, the cells strongly expressed p53 (70%) and MIB-1 (35%).
Discussions: In many reported malignant cases, the histology of the tumour cells suggested that they were malignant, yet the clinical course has been benign. Carefully reviewing the literature, it seems that actually we have enough histological criteria to identify the cases with biological adverse outcome. Those unfortunate cases behave as high grade sarcomas and therefore may deserve an aggressive therapeutic treatment.
  • asked a question related to Cytology
Question
2 answers
For observing chromosome aberration, treated allium cepa root tips were stained with Acetocarmine and destained with 45% acetic acid. But while obseving under microscope, cytoplasm is also stained. Staining time was 3-4 h.
Relevant answer
Answer
Instead of acetocarmine, you can go with "Feulgen staining" which stain only nucleic acid.
  • asked a question related to Cytology
Question
11 answers
While doing cytotoxicity studies of Cyanchum on Allium cepa root meristem, majority of cells were found to be broken. Is there any effect of the biomolecule on it? Which of the phytocompounds (in general) are responsible for cell breakage?
Relevant answer
Answer
Cell breakage describes the phenomenon of cell disruption. In mammalian cells with plasma membrane containing phospholipid bilayers with the hydrophobic lipids in the membrane partitioning outside it would be lysis. Plant cells have very thick walls with chlorophyll and other carbohydrates, and cannot be easily lysed.
  • asked a question related to Cytology
Question
9 answers
We have designed a novel family of drugs that target an enzyme within the mitochondrial matrix.
A number of those compounds have been shown to inhibit the target enzyme in cell-free assays. However, only one compound is active in cell-based assays.
We attribute this discrepancy to the fact that most compounds cannot freely access the mitochondrial matrix.
Which would me the easiest way to test this hypothesis? In other words, we would like to have a rough idea of the concentration of each compound in the mitochondrial matrix. It does not need to be very precise.
Relevant answer
Answer
Hi Edelmiro,
I'm not a chemistry, but i think that you could add to your compound (and as control even to the others drugs that don't work) some specific sequence for a mitochondrial matrix enzyme. Then you could verify that only your drug is it inside the matrix because it will be the only that will give you the specific reaction. In this way you will demonstrate that only your compound is inside the matrix and not the other. Probably is not easy to do, but you could try. On the other hand, you wrote that your compound block an enzyme inside the matrix. You could evaluate some substrate of this enzyme and you should find a different amount ( phosphorylation- cleavage - i don't know what is your enzyme )compared to the untreated or the treated with the un-functional drugs.
i don't know if they are good ideas, but with so few informations i can't help more.
regards.
Francesco
  • asked a question related to Cytology
Question
5 answers
I need some meaningful contributions please.
Relevant answer
Answer
Yes
The HPV virus has to reach the basal cells of the cervix. Therefore, there must be some form of break in the epithelium for the HPV to reach the basal cells. Cervicitis is one way in which there can be a break in the epithelium. A similar situation exists in throat cancer and the relationship between smoking and drinking spirits. The smoking and spirit drinking results in a break in the epithelium and therefore HPV can reach the basal cells. In cervical cancer it does not have to be cervicitis and the process of metaplasia is also thought to be associated.
  • asked a question related to Cytology
Question
2 answers
Cam 5.2 is meant to be a cytoplasmic stain. Apart from the nuclear halo, it does stain the cytoplasm well and also doesn't give any nuclear staining as expected.
Relevant answer
Answer
Thank you Danilo. I noticed this artefact on CK7 staining as well. Might be an effect CK7 has on certain cultured cancer cells.
  • asked a question related to Cytology
Question
3 answers
I have spent some time evaluating cytology extensions from bronchial alveolar lavage, but I do not have enough information regarding this issue, and I can't find any protocols to improve my diagnosis. Can anyone recommend some articles or any book I could consult?
Relevant answer
Answer
We follow the same basic steps in reporting:
1- Adequacy: assessed by many alveolar macrophages (not columnar and certainly not oral squamous). Number is not absolute and depends on method of preparation, but ~50 per slide would be a minimum for me
2- Presence or absence of evidence of viral, fungal or bacterial infection, such as inclusions, inflammatory cells, frothy background of Pneumocystis. etc. Abundance of neutrophils, eosinophils is noted.
3- Presence or absence of Atypical or malignant cells and type.
4- Specific comments in case of possible aspiration, allergy or pulmonary hemorrhage (as outlined in the previous response)
The typing of lymphocytes as to T and B by immuno is not reliable. It should be done by Flow cytometry and needs a separate sample, specifically processed for that, This is unlike FNA, where the samples are much more cellular and can be preserved rapidly, thus preserving the integrity of surface markers.
(see Erozan: Pulmonary Cytopathology, just published by Springer, 2014)
  • asked a question related to Cytology
Question
3 answers
I am trying to optimize assay sensitivity for molecular diagnostics from a fine needle aspirate biopsy, but I'm struggling to find good information on the total number of cells harvested from the procedure.
Relevant answer
Answer
Dear James
This article enclosed may be useful for you
Quantitative analysis of fine needle aspiration biopsy samples
  • asked a question related to Cytology
Question
5 answers
I need more explanation on this, what is the difference between "H &E" and "H+E". In mathematics "and " is refers to as "plus". Ok if it is called Haematoxylin processing, who coined the name. Is it an appropriate name? if yes than other stains will also following, eg. Alcan blue processing etc.
Relevant answer
Answer
Am not aware of the so called new name " H+E" for Haematoxylin and Eosin stains, all articles both in local and international journals carry H&E as an abbreviation for haemtoxylin eosin stains.
  • asked a question related to Cytology
Question
4 answers
I had a difficult time having poor staining quality especially with H & E, Leishmania and rapid cytology stains. We investigated all possible causes without revealing the cause. Then one of the technicians observed some cloudiness or faint smear-like dirt on the surfaces of the new slides.These slides were supposed to be of excellent quality and made by a reputable company! Has anyone had a similar experience?
Relevant answer
Answer
Dear Wolfgang
That was very helpful and at least showed me that these things do occur. I will consider your experience with cleaning the slides. Thanks with my best regards and wishes.
  • asked a question related to Cytology
Question
2 answers
Ehrlichia canis is mostly seen in monocytes, whereas, E. ewingii has been reported in neutrophils and E. platys in platelets.
Relevant answer
Answer
There are many members of the genus and also Anaplasma in animals that infect a broad variety of blood cell types.
  • asked a question related to Cytology
Question
15 answers
A biliary stricture (common bile duct or confluence) without history of calculi is often proposed for exploratory surgery due to low sensivity of bioptic procedures and the need to avoid further bilirubin rise.
Relevant answer
Answer
I suppose that we are considering extra-hepatic bile stricture.
I think that a preoperative cytological confirmation is not necessary. The incidence of benign biliary stricture in absence of previous surgery or calculi is less than 10%. The chance to have preoperative positive cytology is about 50%. In case of a report "negative for tumor cells" are we so confident in suggesting follow-up or stenting? We have to clearly inform the patient, what about: you have a 90% chance to have a malignant stricture.
It's extremely important to have a well done pre-op work up and than we can offer surgery without hesitations
  • asked a question related to Cytology
Question
26 answers
Can you guys list the institutions which provide SEM and TEM service for biological specimens?
Relevant answer
Answer
TNAU having both SEM and TEM facility facility is available.
animal science reserach institute (madras vertinary collage) TEM
facility is available.
IIT(M)also having SEM (SAIF lab)
  • asked a question related to Cytology
Question
10 answers
Which markers? (histopathology, serum enzymes, genes, ...)
Relevant answer
Answer
There are various biomarkers for inflammation in the GI tract. Calprotectins may be used as a biomarker for evaluating in GI inflammatory disorders.
Attached is an article that may be of some used to you when looking for markers of inflammatory conditions of the GI tract.
  • asked a question related to Cytology
Question
3 answers
Or is it of no worth to get it in a Souvenir?
Relevant answer
Answer
I too, have wondered about this! If an abstract is published, in the proceeding of a research gathering, it definitely is worth some credit, in terms of points or else. But, I have not come across any such quantification.
  • asked a question related to Cytology
Question
18 answers
Onion root-tips bathed in suspected water for 48 hours, what was observed appeared to be dense abnormal prophase (B) as well as dense metaphase (A), also abnormal.
Relevant answer
Answer
A is possibly sticky chromosmes at Anaphase and not metaphase as the chromatids are already migrating to opposite poles. B. is an abnormal stage in prophase. For clearity and easy identification of the abnormal chromosomes i suggest that the magnification be increased. Thank you
  • asked a question related to Cytology
Question
13 answers
FISH with Y chromosome-specific probes can be done, but it is time-consuming and relatively expensive.
Relevant answer
Answer
logically, We can use barr body as marker to discriminate between male and female nuclei. histone macroH2A1 staining is used to identify barr body due to histone macroH2A1 content of histone protein which play important role in silencing of whole X chromosome.
Best regards.
  • asked a question related to Cytology
Question
3 answers
Liquid Based Cytology
Relevant answer
Answer
TinPrep: The technique is simpler but expensive. It is less cells then Surepath.
SurePath: More handwork but cheaper. More cells, no problem with bloody and mucoid specimen. You find more adeno ?
  • asked a question related to Cytology
Question
8 answers
Microvesicles (MVs) or exosomes measured 30-1000 nm and all publications noted them through electron microscope but unfortunately EM is unavailable in our institute. Is it possible to visualize the MVs or exosomes using confocal or phase contrast microscopes? I mean to use a special/fluorescent stain for MVS to be easily investigated.
Relevant answer
Answer
Since the size of MVs and exosomes close (or under) the physicial resolution limit of visible light microsocpys, you have to mark the vesicules with fluorescent markers to make them visible - so you will not see directly the subcell. ves.-s, but their fluorescent halo. The fluorescent markers cen be specific (such as fluorescent protein conjugated specific antibodies), or aspecific (using lipid markers, such as Annexin V for phos. serin, or other PKH stains). With the first method you can detect specific population of subcell. ves.-s, while with the other you can detect all of them. It also important to make a control of specificity of your stain, such as using detergent (e.g. Tween, Saponin, etc.), to eradicate the vesicles - in this case you wont see fluorescent marks. A third approach can be using intravesicule markers, as dr. Ganglum wrote.
By the and, let me recomend a good protocol oriented article: Detection and isolation of cell-derived microparticles are compromised by protein complexes resulting from shared biophysical parametershttp://www.ncbi.nlm.nih.gov/pubmed/21041717
With best regs: Csaba Timar
  • asked a question related to Cytology
Question
4 answers
I want to check the vitality of PDL fibers for tooth and dental pulp cells. Are there any strains other than tryphan blue that can be used to check for cell vitality?
Relevant answer
Answer
Besides the usually used Trypan Blue also can be used: methylene blue (absorptive stain), indigo carmine (contrast stain), Cresyl violet; Erythrosine (Red No 3), Eosin (both are exclusion dyes), Propidium iodide; 7-aminoactinomycin D, Alizarin Red, fluorescent dimethyltin(iv)-alizarin red S complex....all depending on the exptl. necessity you have. Even injections of lead acetate as an intravital stain of calcification sites have been described. Perhaps interesting to you also would be reading: Time-Saving Benefits of Intravital Staining
Best wishes and regards,
  • asked a question related to Cytology
Question
14 answers
In lung cancer can we study slide touch print cytology to prove that the underlying lung mass is a malignant mass, its type and so on? No need for biopsy?
Relevant answer
Answer
In all cases we are able to make a correct diagnosis based on cytology alone. In the lung cancer:
1. The use of immunohistochemistry (IHC) on slides and/or cell block .
2. Mutational analysis in adenocarcinoma.
  • asked a question related to Cytology
Question
21 answers
The subjective method involves scoring of colour intensity of stains on micrographs using adjectives like very strong, strong, moderate, feeble and negative and representing these adjective by numbers ranging from 4 to 0 in that order. With this, I feel strongly that analysis of just a micrograph could be different among three individuals. Software imaging method is very expensive and in fact is not affordable for me.
Relevant answer
You can use ImageJ. It is free, straightforward and give good analysis in terms of cell count, area, color intensity, etc.
It has many advantages over Photoshop like when u want to differentiate between cells and fibers having same color, with different shape.
You can download from the following link and the user manual is attached as well.
  • asked a question related to Cytology
Question
4 answers
We are looking for a brand new cytocentrifuge which will be used for cell sedimentation and fixation.
Since this is very new to me, are there any comments or criteria should I have to concern.
Our samples will be blood cells and some epithelial cells. Afterward, those cells will be stained for ICC/IF study.
If any, who is number one in this market?
Relevant answer
Answer
Thanks for your comments. Anyone ever has experience with CellSpin II from Tharmac?
  • asked a question related to Cytology
Question
4 answers
I want to know the cytolytic assay to confirm the presence of cutinase enzyme.
Relevant answer
Answer
Sir it is a bacterial culture
  • asked a question related to Cytology
Question
2 answers
I have some worries about the observation of blades in cytology.
These blades are produced from tonsil samples in ThinPrep and are very rich in lymphocytes. One does not wish to solely have epithelial cells.
I would like to eliminate most of the lymphocytes before observation.
One does not wish to use FACS for cost reasons.
Would you have any idea about the separation of the cells by density gradient, like the Ficoll technique for the PBMC?
Namely because ThinPrep contains liquid of conservation (methanol 50%).
P.S. some examples of observation
Relevant answer
Answer
Hi Babak,
Sorry about delay too.
I see your techniq wich use nylon wool, but this techniq is efficient for purification of lymphocyte. But here my question is the reverse, i wish purificate my epithelial cell and eliminate lymphocyte.
Thank you Babak.
Alexandre.