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COMPENSATION

COMPENSATION. FL1. FITC. +. detector. FITC -. PE. gain=1. -. FL2-%FL1. FITC. FL2. PE. +. PE -. FITC. gain=1. detector. -. COMPENSATION. FL1. FITC. +. detector. FITC -. PE. gain=1. -. PE. FL2. PE. +. PE -. FITC. gain=1. detector. FL1-%FL2. -.

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COMPENSATION

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  1. COMPENSATION FL1 FITC + detector FITC - PE gain=1 - FL2-%FL1 FITC FL2 PE + PE - FITC gain=1 detector -

  2. COMPENSATION FL1 FITC + detector FITC - PE gain=1 - PE FL2 PE + PE - FITC gain=1 detector FL1-%FL2 -

  3. Uncompensated vs Compensated FL2 FL1

  4. COMPENSATION IS INTENSITY DEPENDENT FL2 fully compensated uncompensated partially compensated FL1

  5. COMPENSATE INSTRUMENT USING STAINED CELLS 1. Adjust PMT voltages using unstained cells 2. Adjust compensation for each fluorochrome

  6. Lymphogated Cells for Four Color Single Laser Compensation B A SSC SSC 0 256 512 768 1024 0 256 512 768 1024 FSC -> FSC -> FSC FSC 030695001 030695001

  7. COMPENSATION UP TO FOUR COLORS WITH TWO LASERS R6 R4 R10 SSC APC-CD8 PE-CD4 PE-CD4 R2 R3 R4 R5 R9 R8 R1 FSC FITC-CD45 PE-CY5-CD8 PE-CY5-CD8

  8. R3 R1 R2 R4 R6 R5

  9. A C R3 B D R3

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