Preparing a single cell suspension is a critical step in any solid tissue flow cytometry experiment. Tissue dissection, enzymatic digestion, and mechanical dissociation are three significant steps leading to the degradation of the extracellular matrix and the isolation of single cells, allowing the ...
The process for preparing a single-cell suspension is largely determined by the origin of the cells. Lauren Jachimowicz, application development scientist, flow cytometry at Agilent Technologies, notes that cultured cell lines can readily be transformed into a single-cell suspension ...
L.Z. collected the specimen and prepared single-cell suspension for sequencing. J.H. finished the bioinformatics analysis. J.H., L.L., Y.H. finished the FACS sorting and co-culture assay. M.X., Y.Y. helped with immunofluorescence staining, J.H. and K.C. wrote the manuscript. All ...
The single-cell suspension used for scRNA-Seq was washed twice with phosphate-buffered saline containing 0.04% bovine serum albumin, and the final cell concentration was adjusted to ∼1000 cells/μL. Cells used for flow cytometry were washed with phosphate-buffered saline containing 0.02% bovine ...
A standard method for preparation of single-cell suspensions of mycobacteria is centrifugation, based on the premise that cell clumps are selectively pelleted by gravity, with single cells remaining in suspension42. However, using FCM we found that the ratio of clumps to single-cells was unaffected...
(Solarbio, China) for 15 min on ice. After washing twice with PBS, cells were resuspended in sorting buffer (PBS supplemented with 2% FBS). Finally, cells in 10 μL suspension were counted using an inverted microscope and a hemocytometer. Cell viability was assessed via 0.1% trypan blue ...
Additionally, fluorescence-activated cell sorting was utilized to investigate the relationship between intracellular drug concentration and cell viability. Uptake of Adriamycin in cultured cells was found to be highly dependent upon the method of cell growth (e.g., suspension versus monolayer cultures) ...
Regenerative medicine and tissue engineering aim to promote functional rebuilding of damaged tissue. Comprehensively profiling cell identity, function and interaction in healthy tissues, as well as understanding how these change upon tissue disruption, s
The single cell suspension was washed by two rounds of centrifugation (1300 rpm at 4°C) and resuspended in 1x Tris Buffered Saline. Cells were stained with Calcein AM live stain dye (Fisher, 1:500) on ice for 30 minutes, and were passed through a 40 μm cell strainer (Fisher) to...
epithelial cells were statistically similar to the control digested for four-fold longer time. We also investigated an interval operation format, which involved processing for short time periods, eluting the cell suspension, replacing collagenase in the digestion device, and continuing recirculation. We ...