1. Optical measurement

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

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

5. Identifying Cells By Scatter Light Analysis

6. Excitation- 488nm
Excitation- 520nm
FITC

7. Identifying Cells By Single Color Analysis

8. Spectral overlap
Ex 488nm

9. 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

13. Data Analysis

16. Gate

18. Exampels

20. Nozzel & sheath pressure
Sort Gate
Sort mode

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

23. Yield
sort
sort
abort
Purity