Science method

Cytology - Science method

Explore the latest questions and answers in Cytology, and find Cytology experts.
Questions related to Cytology
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Routine CT Staging is necessary in managing Ca Stomach.
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Yes as mentioned in most of the replies above.
Please see details in some scientific papers such as http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3296804/
--A gastric cancer patient with positive cytology would be stage IV. -------
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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
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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!
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Hello. We do not routinely take this exam. You've already seen this study. Maybe the article can help you.
Vet Ophthalmol. 2005 Nov-Dec; 8 (6): 401-5.
Conjunctival impression cytology in dogs. Bolzan AA.
In which species will you do the cytology ?
Good luck!
in 6s
Anyone who can host me in their lab to learn ICP measurement protocol? - ResearchGate. Available from: https://www.researchgate.net/post/Anyone_who_can_host_me_in_their_lab_to_learn_ICP_measurement_protocol#58c2bd40cbd5c278170e96c3 [accessed Mar 10, 2017].
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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)?
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Hi, 
    It's not mandatory to use FcR blocking reagents while staining with fluorophore conjugated antibodies. You will find lot's of literature supporting this. However there is a chance of non specific binding of antibodies to FcR on monocyte/macrophage, if the antibody, which you will be using is human origin (very less likely), then I believe you should consider use of FcR blocking reagents. If it is from other species eg mouse/rabbit (more likely), then there will be no point of using this. 3% BEA will be fine. 
good luck. Let me know if I can help with this further. 
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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?
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I work with nanos in fish rather than seeds, but I saw that no one had replied to your question so I thought I would send a few suggestions to you. I have seen several talks on seed germination and growth with nanoparticle exposure and both of the methods you listed have been done (see publications below). The best method may be dependent on the properties of your nanoparticles (type, size, coating, etc.), species of interest (water plant, terrestrial, etc.), and environmental relevance (how would your species encounter the nanos in the environment?).
Good luck!
- Melissa
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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!
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Dear Ivan,
I have already found somebody who will co-operate with us. I want to test a human Pneumocystis antibody solution by IHC. Aim is to use this solution for immuno-precipitation for Pneumocystis in pig BALF. The initial steps are the establishment of a functional IHC on human samples and on pig samples.
If I need the mouse sample, I come back to you. Thank you for offering this.
I have passed on your greetings.
Best regards,
Christiane
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8 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
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If cytospin after fixation is not possible:
You can do IF-taining in tube (1.Antibody, washing steps 2.Antibody, DNA-staining ...)
and fix the cell before microscopy on the slide with low melt 0.5%-1% agarose.
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Cytologist, wheat, mitosis   
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Hi, Nader,
Cytogenetic plant (wheat) techniques, namely, Fixation, Pretreatment, and Staining, please, see at site
Godspeed
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the biological characteristics like growth, physiology, cytology, phenology etc.
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climate change effect the physiology of plant
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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!
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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. 
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Attached I send two documents in PDF with photographs of the sample, stained with MGG, another with Pap. Final magnification of 400x.
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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?
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Cytology? VIA/VILLI? HPV testing? Co-testing?
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Thanks for your question - we're not performing any screening for this study, just interviewing women and HCPs who perform testing about their experiences of the test. (The project is in the UK, so the tests the women will have experienced will be liquid based cytology, proceeding to an HPV test only when abnormal cells have been detected.)
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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.
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Dear Qian,
That is a common problem with acinar structures of pancreas and salivary glands which makes immunostaining so difficult. I have worked on both and what helped me was using a prolonged incubation in serum-free protein block (I have used Dako) and background-diminishing antigen diluent from the same source. I am sure that there are similar preparations from other manufacturers, but for pancreas I have only used those two. You may have to do quite a few experiments to establish the working protocol for timing of incubations with protein block and concentration of your prime antibody (after blocking and using this diluent - usually higher than you would use for another type of tissue).  
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anyone can advise the litrerature about methods(cytological,molecular and other ) of reserch orchid mycorriza?
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I have attached some useful papers. Go through it.
Regards
Abhishek Raj
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Comment please 
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Telomeres are the very end of linear chromosomes and can be defined by sequence (in most organisms T-rich on the "sense" strand), these are covered by specific protein complexes such as Rap1-Rif1-Rif2 and Cdc13 (POT1 in humans). This sequence can be extended by telomerase (telomerase also contains an RNA which is complementary to the telomere sequence which helps to recognize telomeres and extend them). Now, subtelomeres are less well defined on a sequence level, it's mostly the region next to telomeres, these can be repetitive in nature and they are often heterochromatinized. Hope this helps!
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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! 
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Sorry I thought you wanted to analyse immune cells in blood. In that case 
Try adding some dextran 40 to your media at about 30mg/L. Heparin can cause RBC aggregation so don't use it in the media that you use in the microfluidic device. You can wash your RBCs in PBS twice and resuspend in PBS with dextran 40. You could also try increasing the shear in the device as well (higher flow rate or smaller dimension in part of the fluid path) to disaggregate the cells. RBCs often aggregate in low shear environments.
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1.History & Clinical examination.  2.Ulcer edges.  3. Lab reports. 4. Cytological examination.
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Skin lesion, lump, or ulcer that do not resolve in 14 days located:
On the tongue, lip, or other mouth areas
Usually small
Most often pale colored, may be dark or discolored
Early sign may be a white patch (leukoplakia) or a red patch (erythroplakia) on the soft tissues of the mouth
Usually painless initially
May develop a burning sensation or pain when the tumor is advanced
Behind the wisdom tooth
Even behind the ear
Additional symptoms that may be associated with this disease:
Tongue problems (moving it)
Swallowing difficulty
Mouth sores
Pain and paraesthesia are late symptoms.
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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.
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Hello Inaki,
in case you don't get your chambers, an alternative is to mark lines on the bottom of your petri dish, e.g. in 0.5 cm distance, can be by a fine pen, or with a scalpell. Or to buy transparent thin foils with such patterns printed, or prepare such foils yourselfe.
When upright illumination, we just print patterns on paper (usually black background + white lines, but can also be opposite) and put that unterneith of petri dish, for counting. Just in case,
Arthur
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What are the main tasks and problems in image processing cytological and histological preparations.
How can help medical professionals image processing cytological and histological preparations.
I'm interested in the publication on this topic.
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Dear Murali, thank you.
This is a good addition to my question.
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I'm interested in the images of cytological and histological preparations. Where can I find a database of images of cytological and histological preparations?
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Dear @Claudia Mazo.
Thanks for your reply.
Vyacheslav.
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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.
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sorry I am not a botany is not my speciality K.
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Hi again. I'm looking for a nice topic regarding the effect of impaired response to apoptosis for our cytology lecture class. Any suggestions? Thanks
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Cytology? VIA/VILLI? HPV testing? Contesting?
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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.
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What do you refer with "large": battery, capsule or tubule? The largest undischarged nematocyst capsule I ever measured is ca. 55 x 9 µm, which is large for a benthic hydroid, but not quite impressive compared with nematocyst capsules found in some Siphonophores [e.g. Purcell (1984) reported heteronemes capsules of ca. 150 µm] 
Kitatani et al (2015) reported tubules of several mm in some Carybdeid species. Have you checked it?
 
Why is a restitution nucleus in meiosis so called?
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Frequently cited in papers on meiosis and meiotic aberrations induced via various agents, what does the term "restitution nucleus" actually connote? How is it different from the normal functional nucleus? Also are FDR and SDR one and the same or do they have any notable distinction (based on either cytological observations of their fates post-meiosis)?
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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''
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Kindly elaborate about the essence of your project. Are your findings against the soul theory mentioned in Geeta or Kriya Yoga techniqs....
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I want to reduce plastic waste and (in the long run) expense and a colleague has previously used cleanable reuseable funnels but I can't find a product or manufacturer. I suspect providers have more to gain by selling huge quantities of single use plastic funnels.
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Many a times we don't get good cellularity to give a clear diagnosis of ovary in cytology. Any advise please?
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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.
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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?
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Dear Mehmet,
Because of the nature of HPV (epitheliotrophic and local infection), you can find the HPV DNA from direct sampling of tumour tissue by PCR assay. The stability of DNA let you detect the HPV DNA from frozen (at -20 degree) or paraffined sample.
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Mustapha OT (1999). Cytological evidence for the affinities of Urginea indica (Roxb.) Kunth and U. nana (Oyewole). AJMNS 1: 223-225.
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OBA TOYIN MUSTAPHA
Department of Plant Biology, Faculty of Life Sciences, University of Ilorin, P. M. B. 1515, Ilorin, Kwara State, Nigeria
The author can be requested for the full form of AJMNS
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I am trying to use a solution for long-term preservation of DNA in cytological samples.
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TENT buffer is used to extract DNA, the composition should be:
10 mM Tris-HCl, 0.1 M NaCl, 1 mM EDTA, 5% [v/v] Triton X100, pH 8.0
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Clinical vs Biological Death; what are the differences and what is the interphase?
  • Death
  • Biology
  • Clinical
  • Evolution
  • Physiology
  • Legal death
  • Physiological Death
  • Psychological Death
  • Emotional Death
  • Body death
  • Cellular Death
  • Cognitive Death
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How do cells determine what size to grow to before dividing?
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What is the function of Alkaloids in living organisms which produce them? It is quite a mystery. We are not taught about it in our university syllabi. Are we missing a big chunk of biology?
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Hi there,
Alkaloïds are secondary metabolites mainly produced in plants. As these molecules don't participate to primary metabolism, they circulate freely into the whole organism. Their role is purely protective against insects and mamals due to their toxicity on non plant specific targets.
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We would like to isolate the cells from imprints on strips of Millipore filter membranes and culture it further. apart from mechanical shaker and vortexing is there any other method to retrieve cells from NC membrane.
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we are working on liquid based cytology in cervical cancer screening would like to work on hpv testing in a larger scale
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The GOI Operational framework recommends VIA for cervical cancer screening till the time an appropriately priced and suitable HPV test becomes available.
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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. 
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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.
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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.
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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.
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Hello everybody.
I like to know why do some leafy plants need high concentration of enzymes to hydrolysis in cytological experiments?
I studied chromosomal number of some landrace Lepidium species and l had to examined more than 70 tests to find suitable treatment of hydrolysis roottips to direct observation via enzymes. I want to know about the reason of this resistance.
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Dear Dr. Roughani
In addition to herbaceous plants, this problem is also present in some fruit trees, and the cell wall is not broken down by the enzymes. In the article I sent, they present a new enzyme digestion protocol that is fast, efficient and flexible.
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Protandim and NRF2 stimulator do that. Any researcher want to add some thing, I cordially invite them. I am very thankful to your suggestion.
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Use epigenetic modulator both DNMT and HDAC like 5-Aza cytidine (500nM) and SAHA (500nM to 2uM) for 24 to 48 hours in combination.
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Is the free radical able to age the cell and oxidative stress? How aging is related with the process? Is this related with the Bio-hacking the age code?
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  • Free radicals are a toxic byproduct of normal cell metabolism. If not cleaned up by antioxidants, free radicals roam the body, damaging DNA, proteins, mitochondria, and telomeres. And what’s the consequence of this oxidative damage? Aging. Most of the theories of aging mentioned above involve damage to one or more of these important structures. Free radicals are especially hard on mitochondria, pulling them into a vicious cycle. Free radicals impair mitochondrial function, which in turn leads to the mitochondria producing even more free radicals. Eventually the mitochondria become so damaged that they are unable to generate enough energy to meet the demands of the body, which can prevent the body from working optimally.
With increasing knowledge of the aging process and its associated mechanisms, scientists are finding novel ways to “biohack the aging code” to help people maintain optimal health over the years and preserve quality of life. What is biohacking the aging code? It means findings ways to –
  • overcome the natural deterioration process that ultimately leads to interruptions in health and
  • protect against anything that might interfere with the body’s ability to function optimally.
The end goal—to help increase a person’s healthy lifespan, also called healthspan. Though many methods are being studied, one promising method focuses on establishing balance within the body.
Why?
Because the body is an intricate system that requires balance—something that largely depends on body input and output. Specifically, what goes into the body influences body output or how we feel and perform on any given day. Interruptions to this state of balance appear to be at the root of the changes we experience with age that often interfere with day-to-day life.
Therefore, to ensure optimal health and performance throughout life requires that you manage inputs to the system to maximize body output; something that can be challenging in today’s world.
Check out the articleBiohacking the Aging Code: You Are Your Choices to learn more about inputs and their effects on your health, performance and behavior.
Some extra knowledge will be appreciated.
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Does anyone have a recipe for Saccomanno fixative (a cytology fixative) for sputum preservation?
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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
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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!
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Simply, from callus appearance. Embryogenic callus white and granuular apperance while the other callus looking yellow and friable.
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the patient is 18 years old complains of weight loss, abdominal distention and menstrual irregularities. Is there a role for FNAC?
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Have serum alpha-fetoprotein (AFP) and beta-human chorionic gonadotropin (HCG) levels been checked?
FNAs in most cases may help. However the size of the mass raises additional risk of torsion.
Laparotomy with biopsy is a gold standard to obtain a definitive diagnosis.
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Differences that may cause a pathologist to identify cancerous cells vs. nomral cells.Is identification depend on the types of the cell that is evolved or not?
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Interpretation of cytology is based on the complex of features, including cellularity, architecture, nuclear features, nucleo-cytoplasmic reatio, presence of mitosis and/or necrosis etc. You are right that identifiation of malignancy in some cases is based on specific features, typical for some cancers. In addition, you can use motelular testing when having indeterminate cytology
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Looking for ways to improve cell block for cytology.
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Why is a restitution nucleus in meiosis so called?
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Frequently cited in papers on meiosis and meiotic aberrations induced via various agents, what does the term "restitution nucleus" actually connote? How is it different from the normal functional nucleus? Also are FDR and SDR one and the same or do they have any notable distinction (based on either cytological observations of their fates post-meiosis)?
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2 answers
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?
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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.
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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?
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I don't have an universal answer. If you have a touch it means you have both tissue sample and imprint taken from this sample i.e. both histological and cytological material whereas the FNA is cytology alone. However several FNA needle passes may be done in different directions so it's sensitivity could be higher than that of core biopsy, and the cell block could be (I would say should be) prepared as well. The question is whether your reference laboratory is able (both practically and legally) to perform the mutational analysis in the FNA material (cell blocks I mean) in the cases it is needed.
Sincerely,
Alexander Shtabsky
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Liquid Based Cytology
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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 ?
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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?
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Dear Ahmed,
apologize if my reply was not quite revealing. I once had a similar problem for my semithin sections and the problem was that semary "dirt-particles" disturbed the viewing as well as imaging/documenting of stained sections. So I decided to do the same... returning ... and got another batch of slides which was ok. Nevertheless I always clean (HCl-& alcohol,washings in distilled water) slides to be used (even if they are taken from an originally packed box). This helps(ed) always not only for correct sticking of the resin sections but also in achieving good staining results.
Have a nice weekend,
best regards, Wolfgang
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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.
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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
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Ehrlichia canis is mostly seen in monocytes, whereas, E. ewingii has been reported in neutrophils and E. platys in platelets.
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There are many members of the genus and also Anaplasma in animals that infect a broad variety of blood cell types.
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Which markers? (histopathology, serum enzymes, genes, ...)
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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.
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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”.
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You don't happen to have the publication available somewhere ... and a photo of Thomas Ashworth?
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Or is it of no worth to get it in a Souvenir?
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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.
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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.
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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
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FISH with Y chromosome-specific probes can be done, but it is time-consuming and relatively expensive.
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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.
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Can you guys list the institutions which provide SEM and TEM service for biological specimens?
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In PSG tech, the TEM studies cost of per sample is 2800/- with tax.
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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.
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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
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I am looking for some databases to apply some techniques for the detection and classification of abnormalities in images of cytological tests.
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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
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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.
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You are correct that imaging software is more accurate. Try Image J, its free and works on many platforms
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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..
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Sorry , Idont no
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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?
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Hi,
ThermoElectron Corp Cytospin 4 CytoCentrifuge and its a good one.
Good luck.
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I want to know the cytolytic assay to confirm the presence of cutinase enzyme.
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What type of microbes u want to perform a cutinase assay?
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I have included the cells in culture, in epoxy resin directly on multiwell plates. To detach, I tried using liquid nitrogen, but the plate was not broken. Do you know a method to separate the inclusions from the plate? Alternatively can you suggest a good method to include adherent cell cultures with epoxy resin without, however, bringing them into suspension so that I can observe them by TEM?
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How to prevent coverslips from sticking to the bottom of six-well plates when removing for mounting to slide?
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Are there any tricks out there that anyone can provide to help when removing coverslips after a staining procedure from a six-well plate for mounting the coverslips to slides? I use very fine tweezers, but still the coverslips seem to stick to the bottom of the wells due to cohesive forces from the buffer or media, etc..
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Michael, All of the suggestions above are correct. I use hooked needle and tweezers to remove coverslips from the culture dish. I'm not sure what is your technique of staining but I thought that it may be useful to post some other tip... When processing coverslips for IF I use a cassette made of two lids of 24 well plates. Inside of such chamber, on the bottom I put thick (2-3mm) filter paper that later on is soaked with water to prevent coverslips from drying. Top of the filter paper I cover with parafilm on which I put my coverslips. Removing coverslips from the well and processing it in the "wet" chamber greatly reduces volume of reagents (like antibodies) needed for staining. In fact I'm able to use as little as 30ul of diluted antibody solution per 12mm coverslip. Also, parafilm is hydrophobic which helps your solutions to stay on the coverslip. I hope that my suggestions are useful. I can send you a pic of the wet cassette that I'm using for IF staining. Good luck with your experiments! Robert
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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
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hi Alexandra
sorry about delay. I have problem with site.
I don't no whole of your process. but i think you can use nylon wool for separation lymphocyte.
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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?
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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,
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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.
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Thanks David and Christoph
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Does anyone have experience with Merk's Neoclear and Neomount solutions? Does this require additional steps after the standard Papanicolaou staining procedure?
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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....
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I'm working with sea urchin coelomocytes.
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Hello Vinicius,
I'm working with coelomocytes of Paracentrotus lividus.
Christopher
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I need some meaningful contributions please.
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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.
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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?
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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.
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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
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Blood collection done with precautions for asceptic conditions will not get contaminated. It is better to use vacuuatainer, and if it is manual method i.e. collecting with syringe and needle and opening the collection vial, then we have to ensure that; there are no direct air currents like fan etc. nearby, use two burners, light them some 10 minutes before and open the vial between the two, use only Sodium Heparin as anti-coagulation factor, and most imp., mix it well and store at room temperature only, set the cultures not later than 36 to 38 hours, and it should yield proper metaphases. Also when you add the sample to culture media see if it is very old, in that case  L-Glutamine can be added to ensure active proliferation.
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Hello everyone,
Is there any solution to reduce considerably the 4T1 mammospheres aggregation.Is there any other agent that can replace methyl cellulose? 
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Please see these papers. They may help you out.
Cancer Letters vol.323(1) 2012 20-28.
Good Luck!
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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.
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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.
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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) ? 
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Try to fix with only 100% of methanol.  PFA (fixative) consist sodium and formic acid which usually causes some shrinkage and/or distortion of the specimen. This might caused the distortion of the nucleus. Most of the staining (e.g: Wright's staining) for thin blood smear (obtained using blood smear or cytospin) requires methanol as the fixative agent.
All the best.
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What are the differences between cytology, cytogenetics, and karyology?
How do they deal with each other?
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cytogenetics :the study of chromosome and the related diseases caused by the abnormal no or structure of chromosome.
chromosomes can be identified by their size,shape and banding patterns such as Q,G,R and C-Banding.
Karyology:the study of the cell nucleus, organelles,structure and its functions.
Karyotype has the formula.such as 46,xy,....
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A 45 years old man with cc of hematospermia from about one year ago. Negative PMHx and normal P/E. Normal sono, PSA, UA, UC and cytology. Negative response to any antibiotic therapy. Whats the next best step?
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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!
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The LBC was developed in an attempt to reduce gaps in conventional cytology promoting the use of cleaner slides, no superpositions of cells or other obscuring elements. This is due to the filter system where only epithelial cells are retained resulting in a monolayer or a thin layer slide.  In this cytology sampling is similar to the conventional one. However, in order to decrease the presence of artifacts, the cytological sample is dispersed in a liquid suspension that after centrifugation, passes through a filter. Cytology is obtained from the suspension and prepared in a monolayer with the morphological structure of the preserved cells. Note that the sample preparation was adequate and correct, if so, you need to differentiate artifacts of cells. The reading of the blade is about the same, the advantage is the precision with which the cytological blade shown. Depending on the site collection, it may be some artifacts that were released from the cells, or technical artifacts. But as for your question, the red cells may be cells that stain of non-specific way, try changing the observation area and see if it repeats the rest. It is difficult to say something when we do not have the vision of the blade.
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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
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Levan et al. 1964 is generally followed. 
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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 ^^
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As our DNA or any other kingdom's or species' DNA is made by same kind of chemistry i strongly believe the DNA repair capacity matters and have a great effect on genomic integrity. The strand breaks (single or double- if they are not repaired) of DNA can be visualized as Chromatid Breaks in the metaphase (by observing Chromosomes). There are lot of publications which can interpret the possibility of gaps and breaks in the chromosome arms (Chromatids) !
1) On the nature of visible chromosomal gaps and breaks. 2004) Savage JR. Cytogenet Genome Res. 104(1-4):46-55.
The above article is just an introduction, if your question concentrates on how the chromatid break happens .?
The damage to the human genome incurs on the order of 1, 000-1,000,000 DNA lesions/cell/day ( Molecular Cell Biology by Harvey Lodish, 4th edition 2000) Most of the lesions are caused by endogenous sources, including reactive oxygen and nitrogen species (ROS, RNS) that can oxidize cellular macromolecules including lipid, protein and nucleic acid where one or the other takes part in the integrity of genome and DNA damage signals
"in common man's words' ...for example..... what does affect the normal cell.?"
the lymphocytes can be affected by the air u breath in
Skin can be affected by the sun light u get exposed
squamous cells in the the passages of the respiratory and digestive tracts, and the linings of the hollow organs of the body can be affected by the HOT DRINKS you take and the SMOKE u inhale.. and they get repaired...this is all normal stories Check out the below article it might be useful
2) Schanz S , Flockerzi E, Schuberth K, Rübe CE (2014) Genetically-Defined DNA Repair Capacity Determines the Extent of DNA Damage Accumulation in Healthy Mouse Tissues after Very Low Doses of Ionizing Radiation. J Carcinog Mutagen 5:196. doi: 10.4172/2157-2518.1000196 good luck!
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3 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 ^^
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Thank you so much Mr. Silva and Emin for recommendation, I am going to download it
then I need your advice later
Thank you :)
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1 answer
cytological studies?
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The nature of chromosome pairing at meiosis in some tuberiferous Solanums (2n, 3n and 4n) is described. A number of abnormalities have been noted at meiosis in the diploids including the formation of multivalents and univalents. Although the occurrence of univalents as early as diakinesis suggested that chromosomal differences do exist, the evidence from the diploids as to the nature of the basic genome in Solanum is not clear cut. An examination of meiosis in triploid hybrids (4n × 2n) revealed a considerably higher value for the mean number of bivalents plus trivalents per cell at metaphase I (14.2; 13.5; and 13.3) than has heretofore been reported. The maximum number of such associations recorded was 16. It appears that in the hybrid between the induced tetraploidS. chacoense (n=24) andS. neohawkesii (n=12), at least 8 out of the 12 chromosomes from the diploid parent are capable of exhibiting partial homology. The analysis of pairing in the triple hybrids ((6n × 2n) × 4n) is more complicated. However, there are some indications that autosyndetic pairing within the complement of chromosomes derived from the 4n parent may exist. The occurrence of pairing within the haploid set of chromosomes derived from one diploid species appears to be suggestive of the possible existence of two subgenomes within the haploid genome but further studies of the whole genus are required for a verification of this possibility.
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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?
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@Please enlist the difficulties/observations you have come across.
Regards
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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?
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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)
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4 answers
cytological criterias are helpful ?
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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.
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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?
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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.
Question
I recently learn vaginal cytology in estrous cycle. as i know, there are abundant of leucocytes in metestrus phase and the amount of leucocytes decrease until disapear in estrous phase. where did those leucocytes go? Are they apoptotic or back to the blood flow? if they're dead, how did the debris gone?
bunch of thanks for response :)
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5 answers
Cervical cytology images contain nuclei,cytoplasm and background. If the nuclei overlap each other what method can be used for segmentation
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Could you post some example images?
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15 answers
Can anybody help me to get the protocol for wright staining? I want to stain the different cell present in sputum. 
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Prepare always fresh specimen- Peripheral smears  from a freshly  drawn blood specimen (EDTA purple top tubes). Allow smears to air dry completely before staining.Please mix the Wright Stain and buffer mixture before blood smears are
made or before the smears are air drying. Combine 15ml of Wright Stain with 75ml of Wright Stain Buffer, mix well and allow to stand for at least 10 minutes. Make sure that a metallic sheen is observed on the surface before staining, Place freshly prepared and air dried blood smears in a manual  staining rack -->Place slide rack with air dried blood smears in Methanol for 30  seconds---> (Adjust timer for 30 seconds--> Take a disposable pipette and flood the Wright Stain on the
appropriately labeled slides. Set timer for 3 minutes. Place oxidizing Wright Stain and Wright Stain Buffer Mixture on Wright stained slides laying on slide rack (Displacing the Wright .Stain off the slides with the pipette filled with Wright Stain/buffer mixture and viewing a metallic sheen on the top of slides.Set timer for 6 minutes. Place slides in Wright Stain Buffer for 1.5 minutes. Set timer for 1.5 minutes. Retrieve slide rack with stained slides from buffer rinse and allow to air dry before examination with immersion oil and 100X objective.Wipe excess stain from back of slides with  methanol soaked gauze, being careful not to wipe off the stained side of the
slides.Place on slide envelope or slide flat/folder and proceed to the microscope room. Place 1 drop of immersion oil onto slide to be viewed making sure
it is completely air dried after staining. 
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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?
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What do you mean by the cells are broken?
 
 
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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.
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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
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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.
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Instead of acetocarmine, you can go with "Feulgen staining" which stain only nucleic acid.
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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.
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Thank you Danilo. I noticed this artefact on CK7 staining as well. Might be an effect CK7 has on certain cultured cancer cells.
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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.
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Dear James
This article enclosed may be useful for you
Quantitative analysis of fine needle aspiration biopsy samples
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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.
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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: