Cytology - Science method

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.

These are the microscopic images of a fruit pulp observed under Binocular microscope at 40x.

Please help me identify the same.

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!

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?

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

Need some help with my methods

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)?

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.

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)?

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.

"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."

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?

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 ^^

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.

I'm used standard squash technique using fuelgen staining.

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.

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.

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

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?

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!

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?

Morphological, cytological, biochemical and molecular tools are used to differentiate Two species. Which is the best and accurate method?

Is it possible to extract DNA for further analysis/diagnosis from Papanicolaou stained cytology slide?

Noninvasive follicular thyroid neoplasm with papillary like nuclear features is a very good topic for discussion.

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!

Many a times we don't get good cellularity to give a clear diagnosis of ovary in cytology. Any advise please?

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?

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.

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.

anyone can advise the litrerature about methods(cytological,molecular and other ) of reserch orchid mycorriza?

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.

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''

Protandim and NRF2 stimulator do that. Any researcher want to add some thing, I cordially invite them. I am very thankful to your suggestion.

Does anyone have a recipe for Saccomanno fixative (a cytology fixative) for sputum preservation?

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!

the patient is 18 years old complains of weight loss, abdominal distention and menstrual irregularities. Is there a role for FNAC?

Looking for ways to improve cell block for cytology.

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. 

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!

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.

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..

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)?

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?

Does anyone have experience with Merk's Neoclear and Neomount solutions? Does this require additional steps after the standard Papanicolaou staining procedure?

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!

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

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! 

Cytologist, wheat, mitosis   

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!

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

Attached I send two documents in PDF with photographs of the sample, stained with MGG, another with Pap. Final magnification of 400x.

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.

Can anybody help me to get the protocol for wright staining? I want to stain the different cell present in sputum. 

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.

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.

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”.

I am looking for some databases to apply some techniques for the detection and classification of abnormalities in images of cytological tests.

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?

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?

Hello everyone,

Is there any solution to reduce considerably the 4T1 mammospheres aggregation.Is there any other agent that can replace methyl cellulose? 

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.

After a classical Ficoll, I spread PBMC (suspended in PBS1X) on Poly-L-Lysin pre-treated and positively charged glass slides with cytospin (200rpm during 5 min). Then I fix them with PFA 4% and cold methanol 100% before immunofluorescence.

Their nuclei look like those of polymorphonuclear cells or are broken (with "streaming DAPI").

Is this distortion known with cytospin (even so I reduced the speed and time of cytospin) ? 

What are the differences between cytology, cytogenetics, and karyology?

How do they deal with each other?

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!

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

Cervical cytology images contain nuclei,cytoplasm and background. If the nuclei overlap each other what method can be used for segmentation

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 ^^

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

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?

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?