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Flow Cytometry (FCM) / FACS Technology Center

Diagram of Flow Cytometry

Flow Cytometry Introduction

Flow cytometry is a widely used method for analyzing the expression of cell surface and intracellular molecules, characterizing and defining different cell types in a heterogeneous cell population, assessing the purity of isolated subpopulations and analyzing cell size and volume. It allows simultaneous multi-parameter analysis of single cells.

It is predominantly used to measure fluorescence intensity produced by fluorescent-labeled antibodies detecting proteins, or ligands that bind to specific cell-associated molecules such as propidium iodide binding to DNA.

The staining procedure involves making a single-cell suspension from cell culture or tissue samples. The cells are then incubated in tubes or microtiter plates with unlabeled or fluorochrome-labeled antibodies and analyzed on the flow cytometer.

Recommendations for Background Control

  • • Unlabeled cells as blank control
  • • Single color and FMO control
  • • Isotype control
  • • Negative cells control

Fluorochrome Selection

  • • Excitation and Emission
  • • Fluorochrome Selection
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Flow Cytometry Tips

  • • Sample preparation
  • • Fluorescence staining
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Flow Cytometry FAQ

  • • What fluorochromes can I use?
  • • What controls do I need?
  • • Do the cells need to be permeabilized?
  • • Do the samples need to be fixed?

Flow Cytometry PPT

  • • Introduction to Flow Cytometry
  • • Flow Cytometry Basic Training
  • • Principles and Application of Flow Cytometry
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Flow Cytometry Troubleshooting

  • • No Staining
  • • Weak Staining
  • • Nonspecific Staining
  • • Unexpected Staining
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