can i use old blood - (Feb/26/2014 )

dear:all

can i use blood after 24hr.preservation in refrigerator for intracellular and surface staining in flow?

if i used frozen BPMCS in intracellular staining what is the precaution specific for flowcytometry after thawing?and should use immediatly after thawing ?can i transport cells in ice books for 15 min.?

kind regard

asmaa

-semsem shaltout-

Not sure. But what if some of cells disrupt? it would produce background noise!!!

-Inbox-

Inbox on Wed Feb 26 18:57:47 2014 said:

Not sure. But what if some of cells disrupt? it would produce background noise!!!

is this for both questions or the 1st only?

-semsem shaltout-

Generally cells should be thawed quickly. As to your last question, transporting on ice should not be a problem.

I just typed pbmc flow cytometry thawing into google and got some interesting results and protocols, such as:

https://www.researchgate.net/post/Is_it_possible_to_do_flow_cytometry_on_frozen_thawed_PBMCs

http://www.altaweb.it/hepacivac/documents/SOP_IU008_v1.1_Human_PBMC_triage.pdf

http://www.biomedcentral.com/content/supplementary/1471-2172-6-17-s1.pdf

and a lot more...Maybe you shoud just read some of them. Hope it helps :)

-Tabaluga-

1st. Disrupted cells could come outside staining medium (Sheath fluid or whatever).

See above links by Tabaluga. Need to follows correct freeze - thaw protocol.

These protocols are for PBMC. In first question when you mention blood, you mean PBMC right?

-Inbox-