Flow Cytometry


Flow cytometry is a laser-based technique that analyzes cell populations in suspension. Flow cytometry analyses can simultaneously detect several biochemical parameters at rates up to thousands of cells per second.

The Influx sorter can detect up to 14 colors with a maximum of 16 parameter.  Forward scatter is collected from a high sensitivity detector unit with resolution slightly below 0.2 um.  The machine can sort up to 4 selected populations.  Sorted cells may be collected into a variety of tubes or plates. The LSR II can detect 9 colors with a maximum of 11 parameters and is useful for analysis only. All parameters are correlated and analyzed simultaneously. Cells can be characterized simultaneously through the detection of fluorescent probes for nuclear ploidy, cell cycle, apoptotic state, expression of plasma membrane, cytoplasmic and nuclear proteins and other cellular characteristics. Researchers save their data and analyze it using the Verity software line for post acquisition data analysis. Both flow cytometers are equipped with digital data acquisition electronics and DIVA data acquisition software. The Vantage can also use its analog system and software as appropriate.

The flow core has been used to study cells from many origins: bacteria, plant protoplasts, plant and funji nuclei, cultured cells, and primary cells from many animal organ and immune systems.

Analysis

  • Apoptotic cell death
  • Cell proliferation
  • Cell ploidy
  • Cell integrity
  • Cell activation and signaling
  • Cell function
  • Cell transformation or transfection
  • Changes in plasma membrane fluidity
  • Expression of plasma membrane surface proteins, receptors, or adhesion molecules.
  • Expression of cytoplasmic enzymes, hormone receptors, and transport proteins

Specific sample preparation information

A table is included to compare the BD Influx sorter and LSR II flow cytometers. This table gives a list of the most common fluorochromes used but others are possible. An experimental set of tubes must include one unstained sample and one sample positive for each individual fluorochrome used in the experiment. You may also need a set of tubes that includes all possible combinations of fluorochromes leaving out 1 color at a time. The machines work best with Falcon 12x75 mm tubes. Cell density for analysis should be 1 million cells/ml with a minimum volume of 300 ul. Dr. King will provide technical assistance for protocol development and data analysis as requested.
Which cytomer should you use?

Post acquisition data analysis

  • Verity Software House WinList 7 and Modfit LT 3.2

Pricing

Service

Price

Influx Analysis and Sorting

$40/hr

Influx preparation

$80/hr

FACS Analysis

$20/hr
(1 hr minimum)

PC Workstation,
software and printing

$20/hr

Consulting on experimental design and data analysis

$20/hr

Instrument Locations

BD Influx sorter
5115 Biomed Phys Sci

LSR II
5115 Biomed Phys Sci