Dear all
During the ISH, I discovered that there are yellow colour crystals on my slides when I placed them in for the colouring reaction (NTMT+NBT+BCIP). What may be the origin of this, and how can I get rid of these crystals, which are also present in the section on the slides?
I attempted a few methods to remove these crystals, but none of them were particularly effective;
including cleaning the slides with only NTMT.
placing the mailer with NTMT and my slide in a warmer at 50 degrees Celsius.
For your kind reference I have added few images.
Thank you
Good day, Vivaan Kk! Hope you are doing well.
Could you provide more specific details on sample preparation? It will make easier to continue search.
Best of luck in your research!
Yours sincerely,
M. Sc. Vadym Chibrikov
Department of Microstructure and Mechanics of Biomaterials
Institute of Agrophysics, Polish Academy of Sciences
Hi Vadym Chibrikov , the brain were embedded in the Slee cryo super glue and sectioning was done using the cryostat. Thereafter, kept the slides for drying, after 2 hours, I started the insitu as follow;
1. awash with PFA followed by PBS wash and then proteinase K treatment.
2. after that again a wash with PFA and PBS, followed by probe weatherization (overnight)
3. Next day, a few washed with SSC and TBST, followed by antibody treatment (overnight)
4. on the third day, a wash with the TBST and NTMT followed by colour reaction (NBT+BCIP+NTMT).
During the 4th step, after keeping the slides for a few hours in the Colour reaction (during these hours I observe the slides multiple times), when I transferred them to a fresh colour reaction solution, I observed this precipitation happening.
Hope this will help
Thank you.
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