Flow Cell Injector Tip Fluorescence signals Focused laser beam Sheath fluid

Forward Angle Light Scatter FALS Sensor Laser

90 Degree Light Scatter FALS Sensor 90LS Sensor Laser

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

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

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

Standard Band Pass Filters 630 nm BandPass Filter White Light Source Transmitted Light 620 -640 nm Light

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

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

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

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

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

Phycoerytherin (PE) Protein Excitation Emisson 300 nm 400 nm 500 nm 600 nm 700 nm Protein

Allophycocyanin (APC) Protein 632.5 nm (HeNe) 300 nm 400 nm 500 nm 600 nm 700 nm Excitation Emisson

Typical Research Cytometer (Coulter 753) (1980s) Detectors $200-300,000 Lasers Fluidics Computers Laser Power Supply

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

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