Cell-Sorting Services at ICBR Cytometry!

Published: August 16th, 2016

Category: All News

Flow cytometry involves the passage of fluorescently-labeled cells through a laser and measuring the light output, thus obtaining counts of cells and quantitative expression information.  It happens at rates of thousands of cells per second.  Cell sorting is the additional capability to select out perhaps a small portion of those cells for further study.  Up until recently, cell sorters have been extremely complex and delicate instruments that required a highly skilled and experienced operator.  This is great for Core labs, but not so convenient to many researchers.  Now some cell sorters offer a very simple setup that most researchers can learn to perform much more easily.

The ICBR Cytometry Core at the CGRC, while continuing to offer cell-sorting services on the typically complex (but still very capable) Aria-II cell sorter, has also invested in one of the modern, simple-to-setup cell sorters, a Sony SH800.  Our SH800 offers three lasers (blue, red, and violet) and up to 6 emission wavelengths, plus forward and wide angle light scatter.  For cell identification, antibodies offer a wide array of potential fluorescent color combinations and may be combined to define a very select subpopulation of cells.  A typical antibody panel could include, for example, these fluorochromes: Sytox Blue (to exclude dead cells), FITC, PE, PE-Cy7, Alexafluor 647and Alexafluor 700.   And there are many other equivalent fluorescent dyes that also work well.

But perhaps the most common cell sorting application is using a single fluorescent label like green fluorescent protein (GFP) as a marker for transfection.  In this way, cell sorting can highly enrich a rare population of genetically altered cells for further study through cell culture and/or genomic analysis.

The Cytometry Core can assist researchers in utilizing the SH800 for their own work.  We would be happy to discuss your application and train you in the use of the instrument.  Please schedule a consultation with the Cytometry Core if you are interested.

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Figure 1: Screen view from SH800 cell sorter, showing CD19-PE (B-cells) on the lower two graphs. The percentage of quadrant “Q1” represents the B-cells, 11.64% of all defined cells.

 

Figure 2: The same sample as in Figure 1, but after sorting for B-cells.  The percentage of B-cells in the sample is now 95.4%.

Figure 2: The same sample as in Figure 1, but after sorting for B-cells. The percentage of B-cells in the sample is now 95.4%.

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