1. Flow Cell Injector Tip Fluorescence signals Focused laser beam Sheath
fluid

2. Forward Angle Light Scatter
FALS Sensor
Laser

3. 90 Degree Light Scatter
FALS Sensor
90LS Sensor
Laser

4. Fluorescence detector
Fluorescence Detectors
Fluorescence
FALS Sensor
Fluorescence detector
(PMT3, PMT4 etc.)
Laser

5. Fluorescence detector
Fluorescence Activated
Cell Sorting
488 nm laser
FALS Sensor
Fluorescence detector - +
Charged Plates
Single cells sorted
into test tubes

6. Optical Filters Dichroic Filter/Mirror at 45 deg Light Source
Transmitted Light
Reflected light

7. Standard Band Pass Filters
630 nm BandPass Filter
White Light Source
Transmitted Light
nm Light

8. Standard Long Pass Filters
520 nm Long Pass Filter
Light Source
Transmitted Light
>520 nm Light
Standard Short Pass Filters
575 nm Short Pass Filter
Light Source
Transmitted Light
<575 nm Light

9. Flow Cytometry Optics Laser Dichroic Filters Flow cell Bandpass
PMT 4
Dichroic
PMT
Filters 3
Flow cell
PMT 2
Bandpass
Filters
PMT 1
Laser

10. PE-TR Conj. Texas Red PI Ethidium PE FITC cis-Parinaric acid Common
Laser
Lines
600 nm
300 nm
500 nm
700 nm
400 nm
457
350
514
610
632
488
PE-TR Conj.
Texas Red PI
Ethidium PE
FITC
cis-Parinaric acid

11. Propidium Iodide DNA Excitation Emisson
300 nm nm nm nm nm
400 nm
500 nm
600 nm
700 nm
DNA PI

12. Fluorescein (FITC) Protein Excitation Emisson
300 nm nm nm nm nm
Wavelength
400 nm
500 nm
600 nm
700 nm
Protein

13. Phycoerytherin (PE) Protein Excitation Emisson
300 nm nm nm nm nm
Protein

14. Allophycocyanin (APC)
Protein
632.5 nm (HeNe)
300 nm nm nm nm nm
Excitation
Emisson

15. Typical Research Cytometer (Coulter 753) (1980s)
Detectors
$ ,000
Lasers
Fluidics
Computers
Laser Power Supply

16. Typical Clinical Cytometer
Detector &
Mechanical
Computer System
Fluidics
$90-120,000

17. The Conventional Microscope
Mechanical
tube length
= 160 mm
Object to
Image
Distance
= 195 mm
Focal length
of objective
= 45 mm