AAT Bioquest

What is the difference between Side Scatter Pulse Width (SSC-w) and Side Scatter Pulse Height (SSC-h)?

Posted May 18, 2020


In flow cytometry, the scattering signals detected at a ninety-degree angle relative to the laser is referred to as side scatter (SSC), which provides information about the internal complexity (i.e. granularity) of a cell. For every signal passing into a detector, there are three characteristics being recorded: height, width and area. SSC-w and SSC-h denote the width and height of the pulse signal detected, respectively. The SSC-h versus SSC-w plots can be used to isolate single cells passing through the cytometer. Non-single cells (doublets, clumps or debris) exhibit different width values (e.g. double the width value as in case of doublets) of single cells whilst the height is roughly the same, hence these cells can be removed from analysis.

Additional resources

Fundamentals of Flow Cytometry

Chioccioli, M., Hankamer, B., & Ross, I. L. (2014). Flow Cytometry Pulse Width Data Enables Rapid and Sensitive Estimation of Biomass Dry Weight in the Microalgae Chlamydomonas reinhardtii and Chlorella vulgaris. PLoS ONE, 9(5), e97269. doi:10.1371/journal.pone.0097269

Leif, R. C. (1986). Practical flow cytometry, by Howard M. Shapiro. Alan R. Liss, New York, 1985, 295 pages. Cytometry, 7(1), 111–112. doi:10.1002/cyto.990070119

Spectral Flow Cytometry

Flow Cytometry Reagents