FSC forward scatter

Fig. 1. Orthogonal light scatter (side scatter, SSC) (y-axis) displayed against forward scatter (FSC) (x-axis) for the author s peripheral blood leucocytes. This is in the form of a dot plot. Each dot represents one cell. A gate has been set on the lymphocyte cluster. If there is any doubt, this cluster can be positively identified using a pan-T antibody (anti-CD3) and back-gating, as described in Section 3.2.2. Data were recorded using a Becton-Dickinson FACSCAN.

Adjust side scatter (SSC) and forward scatter (FSC) on an unstained beads control to put the population of interest on scale. 

Figure 1 Forward scatter (FSC) and right-hand scatter (RHS) from (a) peripheral blood stem cell, and (b) bone marrow harvests.

We perform flow cytometry using the FACSCaliburTM system. Datasets including 150,000 7-AAD positive events are acquired. Gates are set on forward scatter (FSC) and side scatter (SSC) parameters, and the nuclear fraction is gated based on 7-AAD fluorescence (FL-3). Analyses are based on the Alexa-Fluor 488 and APC fluorescence. For the BrdU signal, a cutoff value was defined by analysis of controls that had not received BrdU. 

Fig. I. Two-dimensional light scatter assay. Burkitt lymphoma cells induced into apoptosis by incubation with a MAb against IgM were assayed for forward light scatter (FSC) and 90° light scatter (SSC). Region 1, viable cells Region 2, apoptotic cells.

Fig. 5. Comparison of two-dimensional light scatter assay with subdiploid DNA peak assay. Burkitt lymphoma cells were serum-deprived for 14 d and then assayed for (A) forward light scatter (FSC) versus 90° light scatter (SSC) or (B) cell cycle. Note that although all the cells are clearly apoptotic as shown by the light scatter assay, there is only a small subdiploid peak (SD) present in the cell-cycle analysis, demonstrating that in this case, celt-cycle analysis alone would grossly underestimate the extent of apoptosis present in the cell population.

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