Cell Sorting Protocols

The combination of prospective identification/isolation of bone marrow progenitors and quantitative RT-PCR is a powerful tool to understand the molecular mechanism underlying hematopoiesis. Describes the standard procedures of the murine myeloid progenitor staining for fluorescence activated cells sorting (FACS) and RNA purification methods. - [Read Cell Staining for Sorting of Hematopoietic Stem Cells (HSC) and Myeloid Progenitors]

Protocol describes how monocytes can be sorted from a lysed whole blood sample, and that cytospin preparations can be made for examination by light microscopy. - [Read Enrichment Of Monocytes For Morphological Study Protocol]

Questions and answers about cell sorting. Includes: When should I use fluorescence activated cell sorting over bulk separation methods like panning or magnetic bead separations? Will my cells be harmed by the sorting process? How many cells do I need to prepare to recover 1 X 106 of a population that comprises 10% of the cells? Are there ways to improve sort recovery? etc... - [Read FAQs About Cell Sorting]

Flow cytometry is a widely used method for characterizing and separating individual cells. This basic protocol focuses on: measure fluorescence intensity produced by fluorescent-labled antibodies and ligands that bind specific cell-associated molecules. Includes: Immunofluorescence Staining and Flow Cytometry Analysis. - [Read Flow Cytometry Analysis Protocol]

Protocol for live cell prep for sorting. - [Read Live Cell Prep for Sorting Protocol]

This protocol has been established to maintain the efficient use of the AutoMACS cell sorter during periods of heavy use (e.g., three preparations of splenocytes from 16 spleens per week). Includes: Running the SAFE Clean Program; Clearing Debris. - [Read Maintenance of the AutoMACS Cell Sorter Protocol]

Protocol for surface antigen staining of cells for fluorescence activated cell sorting. - [Read Protocol for Surface Antigen Staining of Cells for Fluorescence Activated Cell Sorting]

Demonstrates the use of the FACSort for sorting based on DNA ploidy or the presence of cytokeratin, and the suitability of the sorted cells for morphologic analysis by microscopic examination. - [Read Sorting Aberrant Cells Using Anti-Cytokeratin or DNA Content for Study of Cell Morphology]

Describes how FACSort can be used to enrich for transfected mouse cells expressing high levels of the human thrombin receptor. The sorted fraction can then be cultured in vivo and reanalyzed 12 days later to show that it remains enriched for thrombin receptor-expressing cells. - [Read Sorting Transfected Cells Based on Gene Expression, Followed by Culture in Vivo]