Demo: Free-floating multi-label immunohistochemistry with tyramide signal amplification

Описание: Companion lecture video that explains background info behind this procedure:    • Presentation: Free floating IHC with multi...  

Link to website with contact info and protocols: https://sites.google.com/view/kru-neu...

This video demonstrates how to do free floating immunohistochemistry on fixed tissue sections. Further, it demonstrates how multiple rounds of tyramide signal amplification - using differently-colored tyramide conjugates - can vastly brighten fluorescence signals of the targets being stained. One last aspect the videos shows is how to mount tissue sections onto glass slides and coverslip those slides.

Time stamps:
0:00 - Intro, section retrieval, and tool prep
3:31 - Endogenous peroxidase inactivation steps
6:30 - Example and explanation of tissue "rinses"
8:25 - Primary antibody cocktail incubation
9:22 - Making and use of 1st secondary antibody working solution
14:32 - Making and use of 1st tyramide conjugate working solution
15:55 - Steps to inactivate peroxidase from prior staining
17:11 - Making and use of 2nd secondary antibody working solution
18:40 - Making and use of 2nd tyramide conjugate working solution
19:50 - Prep for and staining with DAPI
21:34 - End staining, section mounting examples
24:59 - Coverslipping
27:02 - Example picture of triple-labeling