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Cell Loss during staining

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nlazarus

Contributor

Posts: 20

Joined: Wed Nov 13, 2013 7:28 pm

Post Thu Feb 20, 2014 6:00 pm

Cell Loss during staining

I have been having major issues with cell loss during the staining protocol. The biggest loss happens right after the washes following the permeabilization with 1X ebiosciences perm buffer. I have been staining in small 96-well plates but have also tried the 96-deep well plates but have had the same issue. Is thegre something I am fundamentally doing wrong? ALl washes prior to the 2% O/N PFA fix are done at 1600 10mins...after the PFA fix, I wpin 2000rpm 5 minutes. Any input would be appreciated!
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Ofir

Master

Posts: 75

Joined: Thu Nov 07, 2013 12:46 pm

Location: US, CA

Post Thu Feb 20, 2014 6:39 pm

Re: Cell Loss during staining

Hi Nicole,
You may want to look at these two threads:
1. Yield of cells in CyTOF - viewtopic.php?f=1&t=107
2. Where did my cells go??? - viewtopic.php?f=6&t=68

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