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Fig. 5 | BMC Immunology

Fig. 5

From: Effect of cryopreservation on delineation of immune cell subpopulations in tumor specimens as determined by multiparametric single cell mass cytometry analysis

Fig. 5

Cryopreservation effects on cellular viability and cell number recovery in renal cell carcinoma and colorectal carcinoma. a Viability and cell number determined by Trypan blue exclusion. For each tumor type 2–3 million cells per vial were frozen and samples were recovered in 10 mL complete media. Determinations of viable cells/mL are shown on a log10 scale. b Cryopreservation effects on surface marker detection and cellular subset identification in primary renal cell carcinoma. Comparison between fresh and CM1-cryo preserved renal cell carcinoma samples is expressed as histogram overlays of surface markers in CD45+ parent population. Percent positive cells as part of CD45+ population associated with selected cell surface markers are summarized in complementary table. c ViSNE analysis of fresh and cryopreserved renal cell specimen. The viSNE maps are colored based on median expression of selected cell surface markers, the intensity levels are represented by sliding scale. Equal sampling totaling 25,000 cells for both fresh and frozen specimens was used in this analysis. Singlet-live cells, as determined by cell length and cellular nucleation state, were used as the top level population. Clustering was done using the following markers: CD45, CD19, CD11B, CD4, CD8a, CD11C, CD34, CD66B, CD14, CD15, CD44, CD3, CD56, and CD16. CM1 formulation (T1) was selected for fresh to frozen comparison. Expression of cell surface markers in upper left corner is represented by ViSNE regions colored in gradient of red. Differences in ViSNE scatter plots between fresh and frozen specimens directly highlight differences in expression levels of surface markers used for clustering, and are highlighted by red masking

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