Normal human peripheral blood lymphocytes (PBL) were labeled with reagents directed against a variety of lineage and/or differentiation antigens and run on the JSAN, using the AppSan software for data acquisition and then classification into various cells with defferentiated functions. compensation.

• Fig. 1: The four dot plots below show peripheral blood mononuclear cells (PBMC) hemolyzed with Lysis buffer, stained with eight disinct antibody/fluorochrome conjugates, after which the lymphocyte population was gated to analyze the data.

Channels and Dyes:
FL-1: FITC-CD8, FL-2: PE-CD56, FL-3: PE-Cy5-CD19,
FL-4: PE-Cy7-CD28, FL-5: APC-CD3, FL-6: APC-eFluor780-CD62L,
FL-7: eFluor605-CD4, FL8: eFluor450-CD45RA

* eFluor™ is a trademark of eBioscience, Inc.

To monitor expressions of gene insertions, fluorescent proteins are widely used as reporter molecules. Initially, green fluorescent protein (GFP) was the only such molecule available. Today, however, multiple fluorescent proteins including yellow fluorescent protein (YFP) and red fluorescent protein (RFP, sometimes referred to as DsRed) are widely used and enable the use of several reporter molecules in the same experiment, for more complex studies of gene expression and the like.

Fig.2 below shows comparison data of DsRed expressing cells excited by 488 nm and 561 nm lasers, indicating cells excited by the 561 nm laser are more sensitively detected and require less compensation than those excited by the 488 nm laser.