Science topic

Histopathology - Science topic

Histopathology refers to the microscopic examination of tissue in order to study the manifestations of disease.
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In an article regarding IHC usen in myeloid sarcoma there was a report on a positive reaction to neutrophil esterase (Granulocytic sarcoma of the lips: report of an unusual case Badri Srinivasan. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2008;105:e34-e36), but i'm not sure if it is he same (synonym) to chloracetate esterase.
Thanks.
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Histologically, granulocytic sarcoma (GS) undergoes various morphological changes in the cells. It is composed of immature cells of the neutrophil granulocytic series. The infiltration of immature, poorly differentiated cells with round to oval nuclei is the predominant finding. Crystalline, rod-like, intracytoplasmic acidophilic bodies called Auer rods are sometimes seen. With these histological findings, it is hard to distinguish GS from other lymphomas including histiocytic lymphoma, lymphoblastic lymphoma, Ewing's sarcoma, lymphocytic leukemia, undifferentiated carcinomas, and primitive neuroectodermal tumors.
You may refer to the article attached below.
Anti-lysozyme and chloroacetate esterase can be used to diagnose GS.
There are two types of esterases, specific and non-specific. The specific esterase will enzymatically hydrolyze naphthol AS-D chloroacetate liberating a free naphthol compound. This then couples with a diazonium compound, forming highly colored deposits at sites of enzyme activity. This enzyme is usually considered specific for cells of granulocytic lineage. This specific esterase stain, chloroacetate esterase (also called Leder stain), stains neutrophils, neutrophil precursors, eosinophils, and mast cells and confirms the granulocytic nature of the tumor.
On the other hand, the nonspecific esterase activity (alpha-napthyl acetate esterase) is seen in monocytes. The cells of granulocytic series are negative for the nonspecific esterase activity.
So, in the article regarding IHC used in myeloid sarcoma in which there was a report on a positive reaction to neutrophil esterase means that the author is referring to the specific esterase namely, chloroacetate esterase.
Best.
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i want to better understand what are the factors affecting the staining process in histopathology section in the lab.
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Factors such as concentration of the dye, pH of the staining solution, temperature, and tissue fixation are the most common factors that affects dye binding.
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What is the best histopathological stain for translucent collagen
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Mason's Trichrome stain is used to highlight connective tissue fibers in a tissue section.
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After histopathology, for how many years the formalin preserved wet tissues (biological specimen) has to be stored in archival and. Method of disposal of those tissues? Kindly help in this regard . Thank you in advance
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Hi Monika,
If your specimens are medico-legal related, it should depend on your institutional law regarding the specimen ownership and disposal. Concerning specimen integrity, prolonged formalin storage will harden the tissues and cause formalin pigments (artifacts). Should you need the wet specimens in the future, I would advise changing the formalin solution once in a while.
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In affected patients with endotracheal intubation. Good nigth
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Metaplasia
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Hi,
I am starting to work on a liver histopathology image classification project and looking for publicly available datasets. Any guidance related to this will be really helpful for me.
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I think Ankush truth answer .
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It is kidney obtained from a rat with early diabetes mellitus type 1 (streptozotocin model).
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Hi
I think the eosinophilic material in Bowman's space is protein debris. I advise looking for signs of proteinuria such as hyaline casts. Indeed, occasionally within the renal tubules there appear to be small amounts of eosinophilic (proteinaceous) material...
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The strains of broiler in consideration are Marshall, Agrited, CHI
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If that is what you mean, I have only found information on effect on haematology and biochemistry and faecal loads (in broilers) and its antiviral effects.
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Oxylipins play a huge number of roles in animals and plants – including cell-signalling, inflammation and wound response – but did they help ancient microbes ‘invent’ multicellularity? https://buff.ly/3wibP9I
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From Royal Society of Biology RSB discussions
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I need input to calculation of sample size for an agreement study. We will collect several biopsies (n=8) from the uterus of mares for histopathological examination. The aim is to investigate if one biopsy is represenative. How do I calculate the sample size?
Bland-Alman analysis? Or is that only for different analysis?
Thanks in advance,
Cheers Mette
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I don't know that I can give you a good sample size estimate for that. The most straightforward calculation, estimating sample size for detecting discordant scores in a sample, gives an n=1383 with a (assumed) prevalence of 10% at a 95% confidence level and a 5% margin of error with a 0.90 assay sensitivity. This would be quite the undertaking (>11,000 H&E stained slides to prepare and score).
I don't know that this is the proper calculation for your question, and perhaps I am misunderstanding what you are looking to do.
I would consider establishing an arbitrary, large sample size (e.g. n=100) and determining the proportion of discordant biopsies that you obtain. It's going to be difficult to determine biopsy sampling error without being able to analyze the entire uterus.
Some things to consider:
  • If you perform more biopsies from a single uterus, you are more likely to find discordant scores. How many biopsies from a uterus need to have a different score to consider the results discordant?
  • Is this discordance meaningful? Enough to warrant multiple biopsies?
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I'm looking for a software to make panels for histopathology images and wound contraction images. Kindly suggest me a software which can import normal jpg/tiff images and make perfect sized images.
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Shreshtha Gaur….My colleagues use Adobe Photoshop for general purpose work. Those that do specialized image analysis use special software that came bundled with hardware or a variety of open source packages. The most well known and developed is "Image J" or one of the many custom and free variations (download here…https://imagej.nih.gov/ij/) - Image J also has an extensive on-line community that develops tutorials and plug-ins for specific fields of research and reading special file formats. You can read more here…
Another site that offers an overview of some other open source packages is here…
Until you receive other ideas I'd recommend two things; (1) starting with Image J and (2) ask the people at your university what they use. Having a nearby source of help is worth a lot.
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I am looking for suggestions for credible medical conferences for publication on pathology / infectious diseases.
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Please avoid the Rome conference with everything you have. The journals are bad, very bad (even different journal names on the same PDF) and a google search for "is waset.org legit" has lots of reasons to avoid this completely. One headline on an investigation of them: "How the World Academy of Science, Engineering and Technology became a multimillion dollar organization promoting bullshit science through fake conferences and journals." I am not sure if Lotenna is paid by them to promote the conferences or is just naive, but please do NOT get yourself into that rat hole.
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Deep resection for correct histopathological analysis in TUR-B can be challenging for the surgeon with a low to mid-experience level. This leads to inadequate analysis, possibly postponing radical cystectomy.
How do you do it? Do you only rely on what you see?
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Most of the time, it is possible to know if muscle has been included in the specimen from visual inspection and chip thickness. If doubt remains, it is prudent to take biopsies from the floor with cold-cup forceps.
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I have done H&E staining of uterus and ovaries of rats, now want to do scoring of the lesions in percentage%, any idea which technique or software I can use??
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Image J software.
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I need a histochemical method (without antibodies) simple and efficient.
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Movat's Pentachrome.
Pretty complicated.
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I have a number of small bodied fishes that I want to extract gills from, so as to do morphological analayses and histopathology. The problem is: I have a lot more that I need to do the same work with that will be ready in about a fortnight. I don't have regular access to the histology lab so would ideally be able to process all the gills at once. I'm looking to see how I should best preserve the tissue for this period of time? Is that feasible without compromising the sample?
Thanks in advance.
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T@@o provide baseline data on the gill tissue lesions in Huso huso under exposure to direct and water-soluble fraction of diesel oil. In both condition, histopathological analyses exhibited many gill lesions, including epithelial lifting, erythrocyte infiltration, lamellar aneurism, hyperplasia, and lamellar fusion. We report here the results on several abnormalities in the gill structure of the studied fish under direct and water-soluble fraction of diesel oil concentrations which could be used as biomarkers of diesel oil pollution.
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I want to do scoring for macrovesicles and microvesiclular fat diposits in Histopathological Liver sections. Can anyone suggest me a software and discuss a suitable protocol to achieve this. I want to check for the development of Hepatic steatosis or fatty liver condition.
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Previously I used a semiquantitative method for fat accumulation assessment in hepatocytes. the following sentences of our paper (2017) describe the method:" For each section, four to six unbiased counting frames were sampled in a systematic random fashion. Fat accumulation in the liver was graded as previously described (Brunt et al. 1999). Briefly, Grade 0, no fat was found; Grade 1, <33% (light); Grade 2, 33–66% (mild); and Grade 3, more than 66% of hepatocytes affected by fat droplets "
- Brunt E M, Janney C G, di Bisceglie A M, Neuschwander-Tetri B A and Bacon B R (1999) Nonalcoholic steatohepatitis: a proposal for grading and staging the histological lesions. American Journal of Gastroenterology 94 2467–2474.
Hope this will meet your concerns,
Mehran
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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.
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sure, I'd like to see the pictures - philageis@aol.com
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We prepared tissue sections of striatum from excised brains of rats and stained with haematoxylin and eosin (H&E) reagents for the purpose of observing histopathology alterations in the stained tissues. My question now is, can we possibly count the the total number of tyrosine hydroxylase (TH)-positive neurons of substantia nigra in the H&E stained tissues? We would appreciate answers with reasons and references (if available) and any other available guide that could give clarity to the question.
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Thank you Dr Ogunlade Babatunde for your answer. It is much appreciated.
Regards.
Sunday
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Tissue processing techniques in particular..
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Hi Muhammad,
1. Have you placed the bone in fixative (10% Neutral Buffered Formalin or 4% PFA)?
2. Decal in 10% Formic acid
3. Tissue processor protocol for bone
I have a few questions to ask you before I give you specific details.
Please contact me: tina.vanmeter@gmail.com
Regards,
Tina
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How to get access to LYON19 dataset? histopathological image dataset of Lymphocytes provided by LYON19 challenge is not accessible from its challenge site https://lyon19.grand-challenge.org/Background/ .
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Train dataset not available in this chalange..
" No training set is provided, participants should use their own data to develop a method. "
Test data can be downloaded from the  Zenodo platform
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Should I combine both the datasets and train the ML algorithm or should I need to train the algorithm separately with the datasets and test on a single set.The test set is the combination of both the datasets.
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based on your q: " If H and E images are stained from different labs, training and testing with different datasets won't work?? And for hue similarity, can I do color normalization ? "
I would suggest you do colour normalistion to all images as hue would be different. mixed both dataset and divide into train and test so your algo will learn & train on wider variety of image compositions.
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If you have let say a case report that you wish to have published and you submit your paper to a surgical related journal. Do you have to add histology slides?
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In my opinion it's OK to add histopathological features because the reader better understand what the case you are publishing, especially if your case really requires a diagnosis with histopathology
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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
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Good day,
Equipment of Histopathology and Cytology lab depends from volume of biopsies which is planning to be, the clinical departments etc. I can give you some advises about Histopathology equipment, in my country Histopathology and Cytology are different departments.
Best regards,
Dmitry
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recently i have some researches about histopathological changes in the bone , therefore i want to know the typical properation methods to (SEM ) photos . .
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Depends on what are you looking for in a bone. Bone could be mineralized or demineralized, fractured or embedded. I bit more information is needed.
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I want to know if Reinforcement learning or Q-deep learning can be used for analyzing histopathological images for detecting cancer. I know Reinforcement learning is great for scenarios where labeled data is not available but can it outperform a CNN (supervised learning).
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Dear expertise,
i stained frozen section lung tissue sample against virus protein, which should be detect virus infection. i trying to reduce non-specific interaction using serum (5% BSA in PBS) blacking steps to prevent non-specific interaction. its not sufficient in order to control non-specific binding.
i need your help/suggestion to control non-specific interaction in IHC.
Thank you very much for your valuable help/reading.
-N. Stalin
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Dear Nattan,
If you want to show that the antibody specifically detects the viral proteins, an appropriate control would be lung tissue of an animal (or patient) not infected with that virus.
If you use the same staining protocol and microscope settings, you should not see staining in the control (not-infected) tissue, while the infected tissue should give a clear staining.
If this does not work put, you should reduce concentrations of the antibodies, use a different blocking solution (add horse serum, fish gelatine, etc), perform more/longer washing steps and, if the antibody still does not work properly, contact the company. Usually, you can get a different antibody for the same antigen for free, if you show that an antibody showd unspecific binding.
Best,
Sebastian
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Dear Colleagues!
A patient of mine had a tumor of the nasolacrimal duct. CT and MRI scans conclusion was a nasolacrimal duct polyp. I performed endonasal DCR, no neoplasm inside the duct, but the wall was strangely solid and with corkscrew-shaped vessels. I took the lateral wall of a nasolacrimal duct for biopsy.
Histopathological conclusion: a fragment of a mucose membrane, covered with cubical epithelium with mucous glands, with underlying mixomatous stroma, fragments of vessels with thickened walls (cavernous vessels). Morphological pattern is highly suspicious of belonging to cavernous hemangioma.
But: 1) intraoperatively it didn’t look like CH; 2) as far as I know, cavernous vessels are normal for nasolacrimal duct.
Could you help me with the diagnosis? Any cases of CH of nasolacrimal duct in your practice?
Kind regards,
Vasily Yartsev, MD
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Epithelioid hemangioendothelioma?
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I have segmented histopathological image dataset to identify ROI. to check the efficiency of algorithm i need to compare it with ground truth. How can i get the dataset with the available ground truth?
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Hello,
You can find the data in kaggle.com (has a lot of information about several topics) :
and as a extra information here is a post to handle this data:
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The endometrial biopsy was taken in the mid-lutheal phase of the menstrual cycle from a woman with recurrent implantation failure (RIF). Extremely high percentage of stromal and luminal epithelial cells were p16-positive. The observed strong and constitutive expression was significantly different from the p16-expression in the functional layer of endometrium from the other studied patients (more than 200). The studied endometrial tissue was negative for pax-2 and has a “normal” progesterone receptor and estrogen receptor expression. Does anyone have an idea what is the possible explanation for this? Is it possible this p16 expression to be due to a certain pathological condition?
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I'm not a doctor, I am a biologist who worked on cancer previously, and sometimes work on cancer cells now. I was taught that p16 is BAD, and you should have a "heads up!!" reaction whenever you find it overexpressed (see the article). I would check the HPV positivity in the woman, possibly try to get genome data on her (for any unusual cancer-associated mutations, I'm not sure how much and what you can do in your setting) and generally monitor her in the future regularly. Because p16 is NOT GOOD, p16 means "think cancer". So I was taught, at least.
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I am working on hepatoprotective activity. As a part of my study I have to conduct histopathological study of larvae. I went through different research papers. But each paper showing different procedure. Can any one share exact protocol for fixing zebrafish larvae by using 4 % PFA.
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Thank you Melissa Chernick, Daniel Turkewitz, Emmanuel Ifeanyi Obeagu for your guidance,
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IgG4 related diseases are immune mediated fibroinflammatory disease. Hispathologically there are soft tissue fibrosis and lymphohistiocytic infiltrates. Scleroderma is also an immune mediated disease and there are histopathological similarities with IgG4 related diseases.
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There are similarities between IgG4RD and scleroderma. For example, T-lymphocytes seem to be of particular importance in both the diseases. These cells are predominantly CD4+ with a predominant Th2 cytokine profile characterized by high levels of IL-4, IL-5. This key role of T cell proliferation and cytokine secretion suggests that both these condition could be associated with a defective control of T cell activation. B cells are also activated and, through the production of autoantibodies, forced fibroblasts to switch in a profibrotic phenotype. Macrophages in perivascular infiltrates produced transforming growth factor beta that promote fibrosis. However, to diagnose an IgG4RD, histopathological hallmarks must be respected: 1) dense storiform fibrosis and 2) lymphoplasmacytic infiltrate rich in IgG4+ plasma cells (with a definite cut off of IgG4+/IgG+ ratio in the various tissues examined). The IgG4 antibodies in scleroderma are not the most representative and most expressed. In contrast to systemic sclerosis, in which the tissues fibrosis remain largely intractable to treatment, many IgG4-RD patients appear to have a condition in which the collagen deposition is reversible. Therefore, there are some similarities but also substantial histological and pathogenetic differences which must be investigated.
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Hello
My research project involves comparing histopathological changes in Mice Brains from 4 groups. This is my first time working on brain and would like to know how to compare the histopathological changes between different groups, as in, do i have to compare the histopathology from the slides made from same part of brain for all the groups. If so, I have to make sections from the same part of brain right. I am looking for neurodenerative changes to be specific.
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According to my experience in brain sectioning for special staining and brain topography .... you need brain slicer
if you need brain for frozen sectioning.
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Either experimentally or naturally occurring coccidiosis, histopathologic description mentions the presence of schizonts of any Eimeria species in mesentric lymph nodes of any animal species. With references if possible.
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B-lymphocyte responses in the large intestine and mesenteric lymph nodes of mice infected with Eimeria falciformis (Apicomplexa). Nash PV et al. J Parasitol. (1988)
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Noninvasive follicular thyroid neoplasm with papillary like nuclear features is a very good topic for discussion.
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You can work with cytological specimens both retro- and prospective but (see the answer of Olena Polyakova) will need histological (surgical) material to make the diagnosis of NIFTP.
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I have stored the rat's organ at -80°C. How can I do the histopathology of the hippocampus of rats brain? Because at room temperature organ became melted & flexible.
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1. Defrost the hippocampus.
2. Immediately make an incision by the midline in the tissue with a scalpel.
3. Then put the sample in 10% formalin pH 7.2 to fix it for at least 2 hrs.
4. Perform the inclusion in paraffin and continue with the histopathology routine technique ...
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We use very often traditional cryo molds for cryopreservation of tissue samples before cryostat sectioning and we have some inconvenience with that ( not hands free, spend tons of time for that etc.) Probably science went ahead and you use something else for preparing. Could you share with me your experience and I will mutual share my solution for this.with you.
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Freeze a fresh, unfixed tissue sample, up to 2.0 cm in diameter, in OCT in a suitable tissue mold. Freeze the OCT containing the tissue onto the specialized metal grids that fit onto the cryostat.
OCT is viscous at room temperature and miscible with H2O, but freezes into a solid support at −20°C.
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brain tissue slide process
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Your question needs more elaboration. Variations in technique happens according to the need for specific methodology. In addition to Teresa GB's answer I just want to add 30% sucrose solution (for over night) for better sectioning results.
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Dear Scientific Community
What are the Chemicals Companies that can produce highly purified products (Antioxidants and Toxins) with adequate quantities and suitable prices, any recommendations?
Regards.
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1.Sigma chemical company
2.Merck
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Does anyone have access to histopathology images for cancer categorization?
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Thanks, but I need the data for the images you need to process in MATLAB.
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I have been analysing skeletal gastrocnemius muscle specimens from C57 mice and have observed these green rod-shaped structures in the photographs. thinking they were artefacts I ignored them mostly as they were usually surrounding the muscle and not embedded deep within the specimen.
But then looking at the H&E stained tissue, this specimen has a lot more of the rod-like structures placed deep within the tissue and they are surrounded by inflammation. Also they are not staining artefacts as their position is consistent between two specimens cut 5μm apart (see first two pictures).
Has anyone come across anything similar in their skeletal muscle specimens? Tips in how to interpret and understand what is going on would be highly appreciated. I have included other images taken from different part of the muscle specimen to maybe help in interpreting what is going on.
Many thanks!
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I think you could have some hair contaminating these tissue samples. The regular striations could be pigment bodies.
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Recurrent chalazia are seen to harbor adenocarcinoma clinically and on histopathology examination 1.
1. Ref: Shah SIA et al: Concise Ophthalmology Text & Atals. 5th ed. Param B (Pvt.) Ltd. 2018: 27-31
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A very good question. Recurrent chalazion is a risk. However patients under immunotherapy ,recurrent ulcerative blepharitis and with other malignancy or known to harbour the risk should be kept a close eye on. And another practice that should be commissioned is -Every piece of tissue excised from the body mandates a histopathology with proper technique of biopsy.
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Histopathological examination never shows cut nucleus at different level
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There is no mechanism as such through which the nucleus avoids sectioning during microtomy. It is a matter of size of the nucleus of your cell sample and thickness of the section you fix during microtomy that decides how frequently you get sections of nucleus on slide. By the way are these paraffin embedded tissue sections? Which cell types form your sample?
I have done histology of ovaries and ovarian follicles of fish, frog and mouse. I get not only nucleus also nucleolus in histological sections. Also try serial sectioning and number them consecutively, I am sure you will not miss sections of nucleus.
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Findings have shown that an embalming fluid containing Formalin, ethanol and Ammonium Salt go a long way to halt putrefaction faster.
Can this mixture affect:
1. The Forensic results to be obtained from the human sample?
2. Histopathological results?
3. Biochemical Results if any?
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In my opinion you have to look at the single constituents. It depends on the concentration which one is the main effector in the embalming fluid. If the ethanol is of high percentage (96-100%) you will get an hardening effect at the surface, lower penetration-rate. If you use lower concentrations it will rather act as a dilution for the formaldehyde with milder denaturation. Formaldehyde has an optimal ph-milieu for binding to specific functional groups of aminoacids. It may act better in aequous than in alcoholic solution. And the presence of ions can influence the pH, the binding-quality and -preference of formaldehyde etc.
A high ion-concentration may mask the binding-sites for formaldehyde, but may act for itself as dehydrating agent.
In older literature you find many fixative-mixtures, and each of it has a special aim. But I think, they were found by trial and error and the scientific reasons are not documentated well.
I am not clever enough to give a really good answer. - maybe someone else?
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Dear all,
Currently I am working on Alzheimer's disease. In the literature i had noticed the staining of both Cresyl violet and H&E for histopathology studies. what are the things we can differentiate through different types mentioned stains.
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To see neuron/glia in hippocampus, Niissl stain (Cresyl voilet) is the conventional one.
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Burn injury causes an acute stress reaction in body which affects different body tissues in different manners. Is there any comprehensive research articles mentioning histopathological micro & macro features?
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Dr. Sorrentino absolutely right. Also depends of stage and area of burn injury.
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I want to do the spermogramme of mal rats and I need a detailled protocol to do that work.
I want also to do an histopathologic study of genetal organs.
please send me detailled protocols and videos if possible
thanks in advance
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Please be aware that you need detailed knowledge on testes/ovaries histomorphology in order to be able to do staging.
Hope that helps.
Best regards, Geertje
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I am planning to conduct anti-gout study of plant extracts in vivo using hyperuricemic rats. Based on the previous papers, some of them did 7 days and 14 days. So, I would like to ask whether 7 days are enough to study the anti-gout activity of the plant extracts? Is it possible to do the histopathological examination as well?
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.14 days study is enough.it is possible to do histopathological test
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Hi Everyone,
Could you please guide me where I should start to write a code to register 2 whole slide images together? I don't know how I should start coding (I mean according to a pair or a book or what?)
Thank you so much
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Thank you so much for your kind reply. I appreciate your guidance and the list of papers and open sources.
Poupack
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I am conducting research to quantify oral dysplasia of different grades with different IHC stains using new method of qualification.
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Dear Natheer,
Here are some resources:
The two articles I included are freely available online as well.
Hope this helps.
Best,
Kin
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I have two groups of patients. Both of them have CT and Histopathology results. I want to compare they AUROC and PPV / NPV etc values in SPSS ? Is that possible ??
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Hi
1. If the sample size of each group is >30, you can develop two independent T test.
2. Otherwise, non-parametric T test has to be developed.
3. For more information, please refer to:
. Landau, S. and Everitt, B. S.(2004). A Handbook of Statistical analyses using SPSS.
.Howitt, D. and Cramer, D.(2008). Introduction to SPSS.
Regards,
Zuhair
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After staining, i observed that pancreatic tissues are discrete in slide, images are not so clear but in case of kidney tissue i have good image ....I used xylene-ethanol method and stained with eosine-iron hematoxylin.... So is there any difference between soft tissue (pancreas) and hard tissue ( like kidney, heart)? should i follow different method for pancreas tissue?
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I'm not familiar with cardiac perfusion fixation so can you please help me on this regard by providing a authentic protocol ?
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Histopathological section of 8 years old child colon, complaining of chronic diarrhea only.The lesion present many many times in the section.
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Honestly, just as a tip/pointer: Make sure the next time you are requested to make a diagnosis that the "Pathology Department in the Hospital" provides at least adequate (if not optimal!) material for evaluation [means: either reliably processed and stained original slides (tissue sections) or adequate images of the section(s), lesion(s') morphology (low mag,high mag), highlighting the most important guiding structures a pathologist MUST know to evaluate as well as to document / keep records of the found alterations, aiding and finally ending up in a correct diagnosis. And, naturally, without a separate formal request, there should be provided / added additional data regarding the patient's anamnesis, the clinical history / disease course, as well as additional compiled laboratory values. And therefore: I for 100% second the reply #005 as stated by Alexander Shtabsky....But: Thank you for letting the colleagues know what the diagnosis was....
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Do you think this a renal amyloidosis or hyaline glomerulonephropathy?
Because both PAS and congro stain are positive?
I attached the HE staining.
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I think this case is a diffuse form of renal amyloidosis
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Does anyone have any scoring systems for liver and kidney histopathological evaluation?
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For histopathological evaluation in our study, the largest lobe of the liver was fixed in 10% neutral buffer formalin, embedded in paraffin and 5micrometer thick section was used for H&E staining. The scanned images were analyzed and gross morphological alterations could be seen. For an unbiased scoring, the identity of slides were blinded and evaluated (by pathologist) for histopathological lesions (necrosis, fat infiltration, fibrosis and mononuclear cell infiltration) and given a score that ranged between 0 (<5%) to 4 (>60%). The lesions observed fell under following categories viz none (score=0), minimal (score=1), mild (score=2), moderate (score=3) and marked (score=4). Further, the distribution of these lesions were also classified as focal, multifocal and diffuse.
Hope this may provide some help!
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part or tissue of liver and kidney that is subjected to histopathological examination in toxicity studies.
what is the best part of liver and kidney wherein we can observe the toxicity effects of our extract?
Thank you in advance.
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Agree with Dr. Aboumosalam,. In addition, if you worked on small animals like rats and mice, you can take LS of the kidney, this section give you a clear vision about the cortex and the medulla together so, you can detect any pathological features easily. In the liver I prefer to take a piece from each lobe to be sure that the pathological effect is localised or generalized.
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In the lab, this week, we were working with tissues from chicken with (possible) tuberculosis (the samples came ready for processing from another department of the Faculty). But we had some problems sectioning and staining them.
We did our common protocol for paraffin inclusion¹ (since we are starting to work with pathologic samples), and the tissue was still hard as it damaged the microtome blade. We stained the tissues that were possible to cut, but many fell off. Those that stayed, looks really bad, since they are scratched.
Does anyone has any recommendation for processing and sectioning granulomas?
And what about the missing sections during staining?
Thanks!
__________
¹Protocol: Alcohol 70° 1h; Alcohol 80° 1 h; Alcohol 96° 1 h; Absolute Alcohol 1 h x3; Xileno 1 h x3; Paraffin 1 h x2 (the samples stay in the last paraffin bath some additional hours, because the program finishes at middle of the night).
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Please share your experience if problem solved with above suggestions or not
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I have transfected cell line with an inhibitor of a particular gene. that gene promotes tumor growth. Now I want to check whether the inhibitor has caused change in the tumor growth. So can I do this by staining techniques and comparing it with the control cell line?
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You could prepare cell pellets on histological glass slides by cyto-centrifugation. Following centrifugation, cell pellets on slide could be easily fixed in ice-cold alcohol or a mixture of alcohol and acetone (50/50, v/v) for 5 min. Subsequently, the cell pellets could be immunocytochemically stained with antibody to cell proliferation-associated proteins ( i.e., ki-67 or topoisomerase II alpha). The later antibody is more specific to cell proliferation stage, where as the former recognizes cell in their cycling phase.
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I am looking for a research article that has specifically written about histopathological effect of filed cancer in case of major cancer sites such as breast, lung, liver, prostate, colon, etc.
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I am already very deep in literature survey since I am in my 3rd year of PhD. I am currently reading about field cancerization... And since my work focuses on histopathological analysis of images.. I couldn't find any good reference paper with respect to histological changes due to field effect. Hence thought to ask here. Thank you for your suggestion.
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I have one lab that injects TTC before the animal is euthanized.  I have another lab that wants to use it on the heart tissue after it's removed, but they don't know the solution concentration or specifics of how to do that.  Does anyone have a protocol for using it on heart tissue? Thanks!
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Try wrapping the heart in cling-wrap once extracted from the mouse and put it in a container and put straight into dry ice. Use 1% TTC in PBS, incubate the sections of the heart on each side for 10 mins at 37C. After 10 mins flip the sections and incubate again for 10mins. Then put the sections in 10% buffered formation and leave overnight at room temperature. That way you get the best contrast. Without formalin, the infarct area is hard to see.
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78 year lady underwent upfront lap sigmoid colectomy for a bulky tumor. Distal resection margin below promontory with distal resection margin 6 cm. post operative course uneventful. Final histology t3n1 with 2/30 nodes positive. However distal resection margin showed acellular mucin. Would she require resurgent for this or close observation with adjuvant chemotherapy?
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Patient recieved adjuvant chemotherapy and is on surveillance more than 2 years, disease free. No rerescection required
she underwent flexible sigmoidoscopy every 6 months
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I have a number of patients with a quite persuasive clinical picture of malignant melanoma and the histopathology not confirming the clinical diagnosis. I require a revision and additional sections from the pathologist, but the answer is almost  the same. In all these cases I can not blame the pathologists for not doing their job properly. Could some clinical features of malignant melanoma anticipate histopathological changes? How often do you have similar experiences? 
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I can tell you that early diagnosis of cutaneous melanoma on acral skin should rely on clinical pathological correlation. Early lentiginous melanoma on acral sites can be clinically very obvious but histopathologically very subtle. There are a some articles published regarding this issue. At least on volar skin, yes, melanoma can be diagnosed earlier on clinical grounds.
Arch Pathol Lab Med. 2011 Jul;135(7):847-52. Acral junctional nevus versus acral lentiginous melanoma in situ: a differential diagnosis that should be based on clinicopathologic correlation. Bravo Puccio F1, Chian C.
Am J Dermatopathol. 2014 Feb;36(2):142-7. Acral lentiginous melanoma: indolent subtype with long radial growth phase. Kim JY1, Choi M, Jo SJ, Min HS, Cho KH.
Ann Dermatol. 2011 Aug;23(3):400-4. Acral Lentiginous Melanoma Developing during Long-standing Atypical Melanosis: Usefulness of Dermoscopy for Detection of Early Acral Melanoma. Oh TS1, Bae EJ, Ro KW, Seo SH, Son SW, Kim IH.
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I'm trying to analyse images of animal tissues produced with a 3DHistech slide scanner. I usually use ImageJ but have problems fitting large enough images in the memory. Are there any other free alternatives, or tricks that could be used with ImageJ? My analysis is simple, just detecting & measuring circular holes of certain size (adipocytes). The images are like 25000 pixels x 25000 pixels.
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QuPath Open source software for digital pathology
Orbit Image Analysis
Both software are easier than matlab. There cut images into tiles automatically.
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What are types of tissues in these slides of clear cell renal cell carcinoma respectively? Necrosis or tumor stroma?
 Fig1
Fig2
Fig3
Fig4
Fig5
Fig6
Thank you for your help!
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Good day, dear Jun Wang 
Fig1 - it is too low magnification to understand what is situated in left part of figure, but right part is area of calcinosis
Fig2- area of necrosis and blood imbebition
Fig3 - Necrosis and particulary sclerosis of pseudocapsule
Fig4- Pseudocapsule of clearcell renal cell carcinoma
Fig5 and Fig6 - Areas of renal tisue with necrosis, so you couldn`t see any epitelium of tubules and endothelium of vessels. Due to cutting of thin sections there isn`t glomerulus.
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I have the idea about PARP-1 and Caspases which are usually taken as apoptotic markers. But still if anyone can enlighten me with some more idea. It would be great if anyone can share some information about the histopathology in stroke model.
Thanks!
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Thanks Dibbanti and Maria for your valuable suggestions!
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Hi,
I want to do immunohistochemistry of GFAP in brain tissue (coronal sections). And this the first time I am doing IHC of mouse brain tissue. Normally for histopathology we take 5 micron sections, but in articles that I referred for IHC, the thickness is considered 30 micron. Please guide me through if the section thickness matters for IHC.
Moreover, I am going to use GFAP Rabbit mAB, provide me general guidance for AB dilution strategy (how much amount of AB solution I need for one slide (3 sections). 
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If your tissue is fixed with 4% paraformaldehyde , the best thing is to do 30/40 µm floating sections. and perform IHC in culture wells. After DAB, you spread them on super-frost+ glass slides and allow them to dry for 1 night before dehydratation and coverslipping.
With these thick sections, you see better the arborization of astrocytes.
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iam not specialized in histopathology and have some work in this area so, iam in need to identify and diagnose the  pathological changes involved in my tissues.  
so, iam searching for a software or any helping material can aid me to achieve this goal.
thanks in advance
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thanks a lot
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I am trying to find out whether the Michels solution used in histopathology analysis could be used and stored in room temperature. Or what would be the optimal temperature to store this solution.
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images 40X
normal and cancer cell
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Thank you very much
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can anyone give me suggest what happen with this aorta?proliferation, hypertrophy, hyperplasia? i give it NaCl 8%
I have picture with 400x magnification in microscope of aorta with HE staining
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I want to compare with wild type picture :)
Anyway, it looks like medial hyperplasia (between elastin lamina). 
Trichrome stain is helpful to detect elastin lamina.
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In Acute oral tox studies (OECD 423),  histopathological examination carried out on those animals died during the experimental period. But if they died with in 24 hrs then histopathological examination should be carried or not ?
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Thank you so much William for your detailed explanation!!
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I am trying to help rural area for diagnosing wildlife disease by histopathology method. Hence, I am looking for light weight, portable microtome, which can cut at least 5 micron. Does anyone have any suggestion?
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@Adejoke: Do you mean the weight of the microtome? I only found the lightest can cut 5 micron is about 14 kg. This is considered ok. 
@Nabil: I want to find a "portable" microtome for field pathology. Hence, IHC may not be possible.
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Hi, I am trying to incubate a rat pancreatic section with two different antibodies (co-incubation):
1. Anti pancreatic polypeptide (goat polyclonal) - did a solo staining and works on rat's pancreas
2. Anti TTMP (rabbit polyclonal) - supposed to work on rats 
The secondary antibodies that I am using are:
1. Rabbit anti-goat (ab50623) - for anti pp
2. Goat anti-rabbit (ab6717) - for anti ttmp
I have done staining on rat's pancreas section using anti-pp + the rabbit anti-goat secondary and it worked fine. But when I mixed both antibodies (anti pp & anti ttmp), I couldn't get any staining for both antibodies. 
I also tried incubating the primaries separately. I incubated the slide with anti pp first then washed with PBS 3X then I incubated the slide with anti TTMP. Still no staining observed. 
I believe it has something to do with the secondary, maybe? Because if you take a look at the secondaries that I am using, they were raised in the same animal that the other one's anti to. If that is the problem then what should I use? Should I buy a completely different secondaries that have no connection to either rabbit or goat? I browsed the list of antibodies but don't see any other animals aside from rabbit, goat and donkey!
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i agree it might be the secondary, have you tried using donkey anti-goat and donkey anti-rabbit? and block with donkey serum if you are using serum. Donkey anti-goat and donkey anti-rabbit secondaries can be found by lifetechnologies (thermofisher)
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Can we differentiate between Post-kala-azar dermal leishmaniasis and cutaneous leishmaniasis on histopathology?
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