Reliable DSP cell storage is a game-changer for single-cell experiment planning, as shown by Attar et al. 2018 and Jimenez-Garcia et al. 2024. We just demonstrated another practical alternative to fix and freeze samples, such as infectious cells, before sorting for scRNAseq.

Tips: After staining and washing, swap out the wash buffer for the dehydration buffer and proceed with DSP methanol fix and freeze cells. At rehydration, the final resuspension buffer is switched to 2X PBS to maintain cell osmolarity while flow sorting. Sort into a tube with 2X PBS. (6/7)

One might expect paired chain proportions to differ, but the SFFS sample had a 69% paired chain recovery, similar to its fresh F1 counterpart, and a slightly higher proportion of TRA and TRβ chains. (5/7)

T-cell composition was similar for the SSFS and fresh F1 samples, including T-regs. Some unresolved cells were seen in each. (4/7)

The SFFS sample had a 7% decrease in the median genes at 20k reads/cell (blue) compared to fresh samples (red), which tracked with example data from the demonstrated protocol. (3/7)

Our goals were twofold: Test sorting DSP fixed cells in sheath fluid & compare the performance of SFFS cells to fresh samples. Using a standard donor PBMC (F1), we made GEM-X 5’ and VDJ libraries from DSP fixed and sorted T cells to compare with previous data and saw similar cell recovery. (2/7)