![]() The paraffin embedded tissue should be cut into 40~50 μm thick slices first. Sample preparation should be finished as soon as possible.Īppropriate washing, enzyme digestion as well as EDTA treatment should be applied for different cell samples to remove impurities and detach adhesive cells to form single cell suspension.Įnzyme digestion, mechanical dispersion and chemical dispersion can be applied to tumor tissue to obtain single cell suspension. The Basic Principles of Flow Cytometry Sample Preparation Are Listed as Follows. Protocol C: Preparation of Peritoneal Macrophages, Bone Marrow, Thymus and Spleen Cells.Preparation of Adherent Tissue Culture Cell Lines.Preparation of Tissue Culture Cell Lines in Suspension.Preparation of Cells Stored in Liquid Nitrogen.Protocol B: Preparation of Tissue Culture Cells for Flow Cytometry.Protocol A: Preparation of Human Peripheral Blood Mononuclear Cells.Our Protocol of Cell Preparation for Flow Cytometry includes the following parts: In all situations, removing cell clumps, dead cells, and debris is essential to eliminate false positives and obtain results of the highest quality.įig.1 A typical flow cytometry experiment. Care should be used when chelation or enzymatic digestion are used, as these may result in the destruction of the antibody epitope. Many methods are available and may involve enzymatic digestion or mechanical dissociation of the sample. It requires that samples should be dispersed into single cells, but also that the single cells should maintain their inherent biochemical components and biological characteristics. In flow cytometry, the preparation of single cells is extremely important. Solid tissue samples, adherent cell lines as well as tumor samples require further processing before they can be analyzed. The most appropriate samples for flow cytometry are non-adherent cells from tissue cell culture. This technique could be used to characterize and define different cell types in heterogeneous populations, assess the purity of isolated subpopulations, and analyze cell size and volume. Flow cytometry is a widely used approach for analyzing the expression of cell surface and intracellular molecules on per cell basis.
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