I am currently designing a study involving the examination of changes in RNA profiling of PBMCs before and after exercise. To achieve this, we need to collect whole blood samples before and after exercise and isolate PBMCs using the Ficoll isolation method. I am concerned that the RNA profile may be altered during the isolation process. Therefore, I would like to know if there are any methods to fix PBMCs before isolation, to minimize the confounding effects of the isolation process.
Dear Prof. Yusik Kim
Yes. You may refer to the link below.
The protocol outlines fixation of blood and isolation of PBMCs and leukocytes from fixed blood for use with Chromium Fixed RNA Profiling workflow. Storage recommendations for PBMCs isolated from fixed blood and post-storage processing conditions are also provided.
An overview of data derived from fixed PBMCs and leukocytes used with Chromium Fixed Profiling workflow is also shown.
https://cdn.10xgenomics.com/image/upload/v1707937588/CG000721_DemonstratedProtocol_BloodFixation_Cell_Isolation_FixedRNAProfiling_RevB.pdf
Hope it is helpful!
Regards,
Malcolm Nobre
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