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Questions related to Histological Techniques
In the past, I have used freshly-made 4% paraformaldehyde for transcardial perfusion of mice/rats and subsequent post-fixation of whole brains. However, for practical reasons I am interested in...
07 December 2016 10,034 3 View
We have some brain samples that were not fixed prior to freezing, making sectioning and mounting very difficult. Has anyone had any success with thawing a frozen brain so that it can be drop-fixed?
28 November 2016 9,479 1 View
Hello I'm doing a lot of immunofluorescence staining on kidney frozen sections. Currently, I have a lot of sections with a really bad structure whereas previously, sections of the same organ looks...
25 November 2016 10,009 3 View
Anyone have done comparison of ffu assay using ice cold methanol and paraformaldehyde? which is better in terms of getting a clear foci? Any tips to get a better number of foci?
17 November 2016 3,760 3 View
Hello, We're currently trying to stain 50um (or thicker) sections of mounted FFPE brain tissue, with no luck so far. We have been using a 1:2000 dilution of 10mg/ml stock DAPi, with varying...
17 November 2016 3,169 4 View
I am currently trying to dissociate normal human lung tissue that has been frozen @ -80C for about three weeks to set up for MACS sorting and upon my first round I obtained an extremely low number...
16 November 2016 1,917 0 View
I used cold isopentane and OCT to froze my tissues (muscle-TA).
10 November 2016 244 1 View
I want to excise the cerebral cortex and hippocampus but i am afraid i may not get these brain tissues intact to enable me cut them with microtome before embedding in paraffin wax for...
06 November 2016 5,457 2 View
I have to preserve brain tissues of zebrafish for at least one year before I use these samples to do microarry . I heard that RNAlater can do this ,but I'm not so sure. Is there anybody has used...
06 November 2016 2,510 1 View
Whenever I perform an in situ hybridization, black specks appear throughout the tissue within a few hours post mounting with glycerol gel. THey are not present while developing in B4 solution,...
03 November 2016 5,049 3 View
I have done IF for GFAP focusing on hippocampus (CA1, CA2, CA3 and DG). I need to do quantification for GFAP positive cells. Is there any valid method for these?
02 November 2016 9,803 2 View
I am using rat brain tissue for cryosection (Immuno-Fluorescence). It would be helpful if someone can provide your suggestions to preserve the brain tissue for longer duration (1 to 2 months,...
25 October 2016 825 6 View
Bird brains cryosectioned and kept in cryoprotectant in a -20C freezer for ~5 years. It was preserved with acrolein and some sections were used for IHC right away. What can I do with the rest of...
24 October 2016 9,222 6 View
I am requesting your kind assistance in proper steps in harvesting, decalcifying and embedding the knee joint samples, assuming that sectioning and histological staining of the knee samples are...
14 October 2016 5,992 2 View
I have come across to alternative methods to analyse mouse brain cholesterol content, one is staining with filipin and the other uses the Amplex® Red Cholesterol Assay Kit by Life Technologies....
12 October 2016 6,680 2 View
Hi, I have a pelet of cells in agarose and frozen in OCT. I cut it, put it on warm SuperFrost glass slide and let it stand for 1 h at RT. Then I proceed to block my samples but during the...
04 October 2016 6,756 13 View
I need the tissues for the isolation of leukocytes for drug metabolite analysis. How long will they be stable at - 80 deg Celsius.
01 October 2016 3,815 1 View
I forgot to add water in modified davidson fluid while fixing tissues. will it affect the sections or miss some details?
27 September 2016 2,049 2 View
I don't have much cells. I centrifuged and did not get true pellet. Just pieces of white material. Now my cells were in about 6 ml medium. If I aspirate supernatant, they will have some cells....
23 September 2016 4,159 4 View
Hello everyone; I am doing paraffin embedding tissue , and for some time reason I have to stop after xylene step until the next day to continue with paraffin embedding, any media to advise ? thank...
20 September 2016 7,944 5 View
I am enriching breast cancer patient blood samples for circulating tumour cells (CTCs) that i would like to store and work on later. I have read a paper where glycerol is used but no concentration...
20 September 2016 5,463 3 View
Hi ,I am trying to work on pepsin for antigen retrieval for formalin fixed paraffin embedded lung tissues.It did not work when I dissolved the lyophilized powder in 10mM HCl at ph 2.I want to know...
12 September 2016 1,312 1 View
I´m using the Reliaprep gDNA tissue Miniprep System from Promega. I want to know if I should wash the sample or leave it in a buffer or distilled water to wash out the ethanol before using the kit.
12 September 2016 2,907 1 View
I would like to know a reliable method of staining plant neucleolus in lathyrus sativus. I have got some old literature with allium cepa nucelolus staining. Can anyone give me the silver nitrate...
10 September 2016 6,140 2 View