Anatomic Pathology - Science topic

نعم ولكن تحتاج الى عملية بحث واسعة وكبيرة للحصول عليه

The process of fixation is irreversible - I agree to that.

But I am wondering, why you would like to rehydrate the viscera. Wouldn't it be easier to simply 'wash out' the helminths (I suspect that they are in the lumen of the intestines?), then make a cell block of the fluid and then look an it via microscope?

Hello Laura,

First of all, I would say, you should cut your slides from each experimental group at the same thickness (4µm works well for paraffin sections; as long as the section you get at this thickness is transparent ,you can observe the overall cell details well).When the thickness exceed 6µm,you cant see the cells properly (Staining solution may not incorporate well, Specially for staining such as HE and IHC).Another point is the incubation time you keep the slides in haematoxylin or eosin makes the colour different (follow a same incubation time for each staining >I would recommend 1.3min in hematoxylin and 2.30 min for eosin for paraffin sections).you may try to mix the haematoxylin or eosin solutions using a slide glass between each staining to avoid uneven mixing (staining solutions tend to precipitate over time) and try to get rid of water as much as you can after you wash the sections in running tap water after hydration step (water makes hematoxylin or eosin solution become diluted, possibly making the light color in your slides; after staining 75 slides in your automated system , slides may have added extra water to staining solutions)

Hope this helps,

Charith

not all bacterial may be see in slides , but in general we can see the bacterial colonies in infected tissue section, especially in severe bacterial infection

This paper may be useful:

Experimental mammary carcinogenesis - Rat models.

Alvarado A, Faustino-Rocha AI, Colaço B, Oliveira PA.

Life Sci. 2017 Mar 15;173:116-134. doi: 10.1016/j.lfs.2017.02.004. Epub 2017 Feb 8. Review.

Best regards,

Daniela

Hello

We stopped using TUNEL as a demonstration of apoptotic cells a long time ago, mainly as a result of the issues you are finding. The TUNEL assay gives far too many false positives, which can result from anoxia, necrosis or even physical shear damage to the tissue. The assay is difficult to perform reproducibly, even on automated platforms.

We now use cleaved caspase 3 IHC, which is fantastically reproducible, easily automated, easy to quantitate and far less likely to give false positives. The antibody is from Abcam (ab13847).

Cheers

Kevin

Hi Dimitri;

As our lab does immunochemistry and histoenzymatic I can tell you by personal experience that frozen sectioning improves results in many ways; as the tissues are not fixed, epitope retrieval steps are not needed in much cases. And the histoarquitecture is pretty well conserved, endogen eznymatic activity is also conserved.

hope it helped!

Tyler E. Curtis

The elements that would be involved in examples I mentioned would copper, Iron and the compound silica.

Do you think this method could work in some of these cases?

Extramedullary hematopoiesis in soft tissue is not so rare, but extremely rare in solitary fibrous tumor. I've never seen. Please share it by case report if possible.

You are indeed an admirable researcher. It is always hard to deal with such a major constraint. I would say yes, you must keep publishing your work. In that case I think creativity and innovation are key.

I'm not sure about the porcine intestine. For activated microglia in the brain I use the Dako mouse monoclonal antibody CR3/43 which labels HLA-DR-DP-DQ, catalog number M0775.

Also try: ASTM STP810  Cytotoxicity Testing: Prediction of In Vivo Toxicity from In Vitro Tests Published: Jan 1983 http://www.astm.org/DIGITAL_LIBRARY/STP/PAGES/STP30156S.htm

Thanks for your interest and for answering.

The problem is that i need to study an effect of serum without albumin, in order to examine the role of albumin for the effect. That is why i really need to deplete it.

I do know that albumin as a protein interacts with many proteins and there is the "problem" of eliminating some of them but it is a "risk" i don't mind taking :)

I know that Dr. James Smyth from Virginia Tech Carilion used a Compresstome from Precisionary Instruments (www.precisionary.com) to section heart tissues. I see their video on Youtube, it works well and https://www.youtube.com/watch?v=Q0PT37BPles.

Hope you find this useful. Good Luck!

Yiying Z

Hi Claudiu,

here are two recent articles.

Persaud, Dharam and O'Leary, James P., "Fibonacci Series, Golden Proportions, and the Human Biology" (2015). HWCOM Faculty Publications. Paper 27.

Marco Iosa, Giovanni Morone, Fabiano Bini, Augusto Fusco, Stefano Paolucci, Franco Marinozzi, The connection between anthropometry and gait harmony unveiled through the lens of the golden ratio (2016) Neuroscience Letters

612, 138–144

Depending on your interest, you may find many researches on the artistic anatomy of Renaissance, and others applying mathematics to Facebook pictures. :)) 

BR,

Anca

hi my friend, in my opinion if you use a good preservation method with Nathan preparation and centrifugate gradually to obtain a cell button and then cell block, may have a very good results.

greetings. 

Thank you guys very much. I will try your protocols asap

Dear Mitesh,

guessing you are doing Fluorescence Microscopy? (or only BF-LM?)

Perhaps you have seen also a recipe for Thioflavin S staining for Senile plaques (amyloid) to demonstrate neurofibrillary tangles and senile plaques in

Alzheimer's diseased brain tissue sections at http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/MODTHIOF.PDF

Also there is an article, "Binding mode of Thioflavin T in insulin amyloid fibrils" by Groenning Minna, Norrman Mathias, Flink James M., van de Weert Marco, Bukrinsky Jens T.,  Schluckebier Gerd, and  Frokjaer Sven, in:   Journal of Structural Biology 159 (2007) 483–497, stating that there are reactions for two binding site populations (but perhaps 3 binding site populations exist). <<The binding capacity was unaffected by pH, but the affinity was lowest at low pH.>>

You might have also a look into:

<Mechanism of thioflavin T binding to amyloid fibrils> by Ritu KHURANA et al, in Journal of Structural Biology 151 (2005) 229–238. (<<The properties of ThT were studied in aqueous solutions as a function of the concentration.>>  and see also subchapter <Specificity of thioflavin T binding>.

It might be that staining in frozen sections might be a bit complicated and I do apologize not to have any clue for your "lighting up like a christmas tree". Is there any specific reason you use Thioflavin T rather than Thioflavin S for localizing tangles?

Best regards, Wolfgang

Dear Christa

Please find the link https://www.bdbiosciences.com/documents/cd_marker_handbook.pdf

You should be able to find the possible CD markers based on the cell lineage [most of the times]

Interesting! Thank you!

I have performed studies on this particular question, on the origin of the splenic artery, through injection of Metacrylates and moulding of the arteries.

I'll have to consult wide bibliography on the subject and post it soon.

Great question, Andrei. Thank you!

I think we cannot assume keloids appear "spontaneously", the most probable reason for these aparent idiopatic cases is minor trauma, mostly unnoticed by the patients

Sara, in addition to the photographic records, in Guanajuato there is a museum where mummies from the local cemetery are exhibited, mostly from around the same period as the post-mortem photographs. There are some "angelitos" in the collection.

Here is a web site with more information:

And here is an article that mentions the "angelitos" and their connection to the photographs:

Some infants were buried with little brooms with which, according to local tradition, were to be used by the "angelitos" to sweep the clouds in heaven.

Attached is a recent photo (from http://www.momiasdeguanajuato.gob.mx/img/8.jpg).

It doesn't make a difference except in how you prepare your 10mM citrate buffer.  Citric acid anhydrous has a molecular weight of 192.12 and monohydrate has a molecular weight of 210.14. So if you use 1.92g of anhydrous / 1000ml to make 10mM buffer, you will use 2.10g of monohydrate / 1000ml to make 10mM buffer.

Also, citric acid monohydrate will convert to the anhydrous structure when heated, like when you do antigen retrieval in a pressure cooker under high heat.

I've always liked the OMNI - TH models.  Here's a link for the TH115:

Abhijeet, thanks. How about cytoplasmic inclusions containing loads of membranous materials in lining eipithelial cells of ileum treated with Salmonella cell-free toxic filterate?

I agree with Viktor, this is the most common used recommendation beside the one, that you need at least 4-5 in 8 serial sections . However, a close correlation with clinical information (especially application of immunsuppressive drugs, which can cause apoptosis as well) is always needed.

Dear colleagues, concrete autoptic strategy is defined by the case being examined in itself. Which method or protocol is the best one? Simple reply: none. Use the benefits of various approaches, combine them and be creative, be curious and be prepared for everything.

If I understood your question correctly, you are asking about inter-rater concordance / reliability (e.g. if the test is positive, what is the rate of agreement between two or more raters?). There are several methods to evaluate this construct, being the Cohen's kappa coefficient one of the most widely used.

For more information: http://en.wikipedia.org/wiki/Cohen's_kappa

There is also an 'International Journal of Anatomical Variations', which has the aim to continue the 'Encyclopedia of Human Anatomic Variation' by Ron Bergman. It is dedicated to human anatomical variation.

I think imaging and pathology are always going to be linked, to what extent remains to be seen. Virtopsy is a great supplementary tool to the traditional autopsy. Will it replace the traditional autopsy completely? I'm not sure. Will it change the way cases are approached? Absolutely. It's not just a simple CT scan and then you're done as many people say. It is very meticulous series of analyses involving multiple imaging modalities (CT, MR, and laser scanning) as well as selected biopsies and biochemical panels. While I recognize that it's a major concern for pathologists that radiologists are going to think that the pathologists are suddenly unnecessary, I don't think it's a legitimate concern. As Michael said, there are things that a normal autopsy picks up that can't be seen in imaging. However, wouldn't it be better if we were smarter about our traditional approach because we were informed by the imaging? We do this in surgery all the time with image guided pre-operative planning. Simple X-ray can't pick up the detail that CT or MR can even in the deceased. Yes, the big concern is the matter of access to the resource & funding but I think as proof of concept is established more here in the US that you will see more and more ME's offices using advanced imaging in the course of their normal case procedure. As a very wise pathologist pointed out to me during autopsy, the traditional autopsy hasn't changed much in hundreds of years. It's about time that technology helps serve as an additional tool as it does in the rest of clinical medicine. If for no other reason but long term digital evidence preservation, Virtopsy is a very useful tool to the ME. Not only can you preserve the data but you can also "share" or collaborate on that autopsy much like is routinely done in clinical telemedicine. I'd be very cautious against throwing the concept or practice out altogether. Regardless, it will be interesting to see where it goes.