I am performing FACS on lymph node cells on Friday. But a scheduling conflict came up with the machine. Now, I must wait 3 hours before I can run the samples even though I must sacrifice at the same, early time (this is a sensitive 72hr-after injection experiment).
My question is are cells more viable in their organ form or in single-cell suspension? I plan to do one of the following:
a) dissect the lymph node from the mouse and place whole organ in cell media (RPMI+10%BSA) on ice?
b) dissect the lymph node from the mouse, process the cells into single-cell suspension (RPMI+10%BSA - same as above), and place on ice?
Either way, these will stay on ice for about 3 hours before I start my protocol. What do you all suggest? Or is there another method more preferable to keep the cells happier?
Hi Thomas,
If I were you, I'll choose the second way, making them into single-cell suspension. If you keep them as whole organs, you'll still make them into single cells before your protocol, right? Besides, I'll not just put them on ice, instead, I'll rotate them in a cold room with 20% FBS.
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