Flow Cytometry

Flow cytometry is a laser based, biophysical technology employed in cell counting, sorting, and biomarker detection and protein engineering, by suspending cells in a stream of fluid and passing them by an electronic detection apparatus. It allows simultaneous multi-parametric analysis of the physical and/or chemical characteristics of up to thousands of particles per second. SBH Sciences houses a BD Accuri™ C6 flow cytometer to assist us with your research needs and questions.

The system is equipped with a blue laser and a red laser, two scatter detectors, and four fluorescence detectors with interference filters optimized for the detection of FITC, PE, PerCP-Cy™5.5, and APC.

Flow cytometry can be a useful tool in many areas of research: immunology, cell and cancer biology, and microbiology. Additionally, Flow cytometry provides a powerful method for investigating cell signaling because the cells do not need to be lysed which is a prerequisite for many techniques. Researchers can first treat cells in a myriad of ways, then fix, stain with conjugated antibodies against cell surface and/or intracellular proteins (permeabilization required), and analyze the cells using flow cytometry.

SBH Sciences scientists also have access to the Attune Nxt Acoustic Focusing Cytometer (Invitrogen) located in the Innovation Lab at ABI-LAB II. This flow cytometer has four lasers, blue 488nm, red 635 nm, violet 405 nm and yellow 561 nm and can perform compensation for your multi-fluorochrome experiments. A recent addition to the Innovation Lab is the Attune CytPix Flow Cytometer which only has two lasers , red and blue, but has a high-speed brightfield camera which captures sharp images of your cells as they pass through the flow cell helping you analyze only single cells and not doublets or debris. Below left is a histogram from a FACS analysis of C6 cells stained with 5 ug/ml rabbit anti-rat IL-1beta. An incremental increase in mean fluorescence was detected as the antibody concentration increased from 1 ug/ml to 5 ug/ml (table).

At SBH Sciences we have also performed FACS analyses to assist clients with their cell cycle, drug testing and quality assurance experiments. The histogram on the right is from Jurkat cells stained with PI/Rnase staining buffer. We also have detected numerous cytokine targets both with intracellular and surface staining and obtained high signal to noise ratios. Please contact us if you would need assistance with your FACS analysis experiments.