| i. NOTE: DO NOT WASH EMBRYOS AFTER THIS STEP!!!
12) Incubate embryos in a 25-50 µl drop of Hoescht 33342 or PI for 15 min at room temperature in the dark.
13) Wash the embryos 7-8 times in PBS/PVP as described in Step 3.
14) Add 2-3 µl of mounting medium (w/ antifade) to a poly-L-lysine coated slide.
15) Transfer the embryos (5 to 20 embryos per drop) and cover the sample with a cover slip.
16) Determine the number of fluorescent nuclei. Blue (Hoescht) or red (PI) fluorescence indicates non-apoptotic cells and greenish-blue or teal (Hoescht) or green/yellow (PI) fluorescence indicates apoptotic cells.
HINT: If background fluorescence is a problem, wash the embryos for a longer duration in Step 13. It is crucial that the non-specific PI is washed away from embryos prior to mounting them.
 Procedure for Performing TUNEL Reaction for Embryos Affixed to Microscope Slides
NOTE: The procedure is written for PI but can also be used with Hoescht instead
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