Flow cytometry is a brilliant tool for exploring the individual characteristics of cell lines and tissues.
What is flow cytometry?
- Flow cytometry is a method to perform rapid, multi-parameter analysis and characterization of individual cells or particles.
- Measurements are taken as single cells or particles pass either a single or multiple lasers.
- These cells or particles are evaluated for visible light scatter and fluorescence.
- Light scatter can be used to determine size, granularity, and even distinguish individual populations in certain cell systems.
- Fluorescence can be used to evaluate viability, surface and intracellular protein expression, and distinguish individual populations based on phenotype.
How does flow cytometry work?
Flow cytometry is an immunostaining method that utilizes the protein/antibody interaction. Antibodies (primary or secondary) are conjugated to a specific fluorochrome (i.e., FITC, APC, PE, etc.; Fig 1). These fluorochromes are excited by one of the cytometer lasers at a specific wavelength and will emit at another wavelength which will be read by the cytometer.
Figure 1. Diagram depicting binding of fluorochrome-conjugated antibodies to their respective surface receptors and how these populations can be distinguished by flow cytometry.
Why choose flow cytometry over other protein analytical systems?
Table 1. List of advantages to using flow cytometry and how western blot and immunohistochemistry (IHC) techniques measure up.
*This comparison is based on the benefits of flow cytometry. Western blot and IHC are both useful techniques in other applications.
What are some practical applications of flow cytometry?
- Verification of cell lines using surface immunophenotyping
- Confirmation of proper differentiation following induction experiments
- Quantitative assessment of GFP or other overexpression/knockdown target transfections
- Characterization of dissociated tumor cells
- Measurement of a target’s fluorescence intensity
- Quantitative evaluation of surface receptor density
- …and many more!
Cell Sorting
Flow cytometry is a wonderful method for exploring cell lines and tissues, but it is not limited to providing just analytical data.
What is cell sorting?
- Cell sorting is a method that utilizes the same principles of flow cytometry but provides pure populations of live cells “in hand” to work with.
- These cells are distinguished and sorted by scatter and/or fluorescence.
How are cells sorted?
- Cells are sorted in a sterile manner and can be used for downstream applications that require aseptic technique such as culturing or CFU assays.
- Cells can be sorted two ways:
- Bulk sorting
- Single-cell sorting
What are some practical applications of cell sorting?
- Bulk sorting:
- Purify a population of interest from dissociated tumor cells, including rare populations
- Select for GFP+ transfected cells to create a pooled cell line
- Sort to exclude unmanipulated cells from an engineered cell pool
- Single-cell sorting:
- Perform easy limiting-dilution assays
- Dispense cells to generate clones of GFP or overexpression/knockdown target transfections
- Sort a specific population for single-cell technologies, such as RNA- or DNA- sequencing
- Measure the fluorescence intensity for targets of interest for each individual, sorted cell
- …and many more!
All of these capabilities along with cell culture services are available to you at ABS Bio. Please contact us to design a study that will meet your experimental needs.
VLADISLAV GOLUBKOV, PH.D.
Research & Business Development Director, ABS Bio
30+ years in life sciences
Biotechnology expert
45+ peer-reviewed publications
Vladislav Golubkov, Ph.D., is Research & Business Development Director at ABS Bio. He is a biotechnology expert with over 30 years of industry and academic experience in biospecimen solutions, biobanking, cell and molecular biology, and translational research. Dr. Golubkov specializes in the commercialization of custom biospecimen and research services, partnering with pharmaceutical and biotechnology companies to deliver high-quality, fit-for-purpose solutions for drug discovery and clinical development across diverse therapeutic areas. He has extensive experience aligning scientific operations with client and market needs to drive revenue growth and long-term partnerships.
Prior to his current role, Dr. Golubkov held scientific and leadership positions at the University of Southern California (USC), Sanford Burnham Prebys Medical Discovery Institute, and Cureline Inc.
He is a published author of over 45 peer-reviewed articles and a former NIH-funded principal investigator.
