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BioForum _ General Lab Techniques _ Isolated nuclei and trypan blue

Posted by: squallweathered Jul 2 2009, 12:43 AM

Hi all,

Long time listener, first time caller!

I'm currently isolating nuclei from HeLa cells and many protocols say that they "confirm the nuclear isolation and purity using trypan blue". I have used trypan in viability assays in the past to count blue/pink cells, however, what should isolated nuclei look like under trypan and how would they look different from whole cells?

Many thanks!

Posted by: bob1 Jul 2 2009, 04:14 PM

They will look just like whole cells, only a lot smaller. The trypan blue should still be excluded by intact membranes.

Posted by: squallweathered Jul 4 2009, 09:42 AM

QUOTE (bob1 @ Jul 3 2009, 01:14 AM)

They will look just like whole cells, only a lot smaller. The trypan blue should still be excluded by intact membranes.

Thanks for that! I'll take a look ..........