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Questions related to Histological Techniques
I have been recently trying to study mitochondrial structures in our cell line B16. Previously we used the Mitotracker Deep Red dye for this study and it worked fine. But in the last few...
07 July 2016 3,952 4 View
I am studying the structure of cmv in lung tissue using Fluorescent Immunostaining but I get a lot of background autofluorescence (autofluorescence seen on FITC and Tex Red channels). Has anyone...
06 July 2016 7,480 2 View
I have been doing a lot of total RNA extraction from wild type zebrafish. However, with other experiments going on simultaneously, I often find very little time left for dissecting the fish and...
06 July 2016 8,169 4 View
i shall be staining the extracted brain after i am done with all of my experimental groups. I shall be storing them for that long. what condition and solution will be optimum for this type of storage?
01 July 2016 9,977 3 View
If yes, could you please advice a protocol? Thank you in advance for your kind help.
30 June 2016 9,461 2 View
The cryostat that I have available to me can only section up to a thickness of 60um. Has anyone done successful spine analysis on sections of this thickness?Also, the slides that I want to use to...
29 June 2016 5,018 1 View
I blocked and mounted a brain for sectioning in a cryostat, but for some reason it didn't freeze after more than 30 minutes, despite being put at -25 C. Normally the brains I section freeze within...
29 June 2016 9,201 2 View
dear all, i would like to ask how to count using allred method in IHC, i have several paper that i have read but i have difficulties in understanding the calculation of intensitiy score, if anyone...
28 June 2016 1,675 2 View
Sucrose acts as a cryoprotectant by removing the water (by exosmosis?) Does this alter the size of the cells? Technically the cells should shrink due to the 30% sucrose hypertonic solution being...
28 June 2016 4,361 3 View
I have been using Alexa 488, 555 and 647-conjugated secondary antibodies and phalloidin, and except for the price, I’m happy with them. But as time passes I am wondering if there is anything...
27 June 2016 4,493 2 View
Using rat brains; kit says to rapidly freeze brains after initial staining protocol and then section on cryostat. Have too many brains to realistically slice in one day. Once frozen, can brains be...
27 June 2016 3,771 2 View
Hi there, Yet another IF-related question. I'm going to be dual-labelling free-floating tissue sections with fluorescent secondary abs. After the final wash steps, I will be mounting sections...
23 June 2016 7,803 3 View
Hello, I am looking for signs of compromised mitochondrial function induced in the retina after in vivo injury. Is there a target for IHC or any other test that can be used to effectively quantify...
22 June 2016 5,788 5 View
Usually i'm reading that cryosections should be stored at -80 or -20? Although if tissue was fixed with paraformaldehyde before processing for cryosection, the slides might be stored at 4 degres....
22 June 2016 5,157 4 View
Having a hard time getting a decent slide out of rat spinal cord sections ive mounted in paraffin. There's tiny streaks/tears in the section it self but not the surrounding wax. Here's the...
21 June 2016 1,156 2 View
some month before i preserved tissue in trizol for RNA extraction purpose but now i want to use it for Immunohistochemistry?
21 June 2016 8,234 2 View
What is the function of sucrose in lysis buffers for isolation of intact nuclei? I observed that nuclei pellets resuspend easier in buffers without sucrose, and have a higher tendency to clump...
15 June 2016 8,536 10 View
I'm looking for a good immunohistochemistry antibody for the dendrites of inhibitory neurons in mouse hippocampus. Ideally I'd like a "broad-spectrum" marker that would mark the dendrites of most...
06 June 2016 447 4 View
06 June 2016 1,398 4 View
Immediately after brain isolation hippocampus was isolated and with the help of tissue chopper 300 micron thick slices were prepared. The slices were incubated at 32 c in ACSF for about 60...
06 June 2016 508 8 View
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06 June 2016 3,158 2 View
I have a batch of mice samples fixed in neutral buffered formalin, and they are for immunochemistry and histology purpose. How can I properly keep them for 3 months or longer?
06 June 2016 4,552 6 View