Optical measurement

Forward Scatter FSC Detector Laser Beam Original from Purdue University Cytometry Laboratories

Side Scatter FSC Detector Collection Lens SSC Detector Laser Beam Original from Purdue University Cytometry Laboratories

Identifying Cells By Scatter Light Analysis

Excitation- 488nm Excitation- 520nm FITC

Identifying Cells By Single Color Analysis

Spectral overlap Ex 488nm

Compensation Fluorochromes typically fluoresce over a large part of the spectrum (100nm or more) Depending on filter arrangement, a detector may see some fluorescence from more than 1 fluorochrome. (referred to as bleed over) You need to “compensate” for this bleed over so that 1 detector reports signal from only 1 fluorochrome

Data Analysis

Gate

Exampels

Nozzel & sheath pressure Sort Gate Sort mode

Interrogation Charging the stream Droplet formation Electrical plates Attraction Waste aspirator Collection

Yield sort sort abort Purity