Flow cytometry is a laser- or impedance-based technology used to analyze physical and biochemical properties of cells as they flow in a fluid stream through a beam of light. It allows for measurement of cell phenotypes and functions by detecting light signals from cells suspended in a fluid stream and passing through an electronic detection apparatus. Key components include fluidics to flow cells, optics to direct light, and electronics to analyze light signals and characterize cells based on properties like size, granularity and marker expression. It has various applications like cell counting, sorting, biomarker detection, and analysis of properties such as cell cycle, apoptosis and calcium flux.

1. FLOW
CYTOMETRY
DIVYA BANU
CRI (2015-2016)

2. WHAT IS IT
FLOW CYTO:
CELLS
MEASUREMENT :
METRY
DEFINITION
Laser or impedence based,biophysical
technology employed in cell counting,cell
sorting,biomarker detection and protein engineering
by suspending cells in a stream of fluid and passing
them by an electronic detection apparatus.

3. HISTORY
•FIRST FLUORESCENCE
BASED FLOW
CYTOMETRY DEVICE :
1968 : WOLFGANG
GOHDE
•ORIGINAL NAME OF
FLOW CYTOMETRY :
PULSE
CYTOPHOTOMETRY

4. PRE REQUISITES
• CELLS IN SINGLE CELL SUSPENSION
• FLUORESCENT PROBES
• CYTOMETER

5. WHAT DOES IT DO ??
• ANALYSES LIGHT SIGNALS TO DETERMINE :
PHENOTYPE AND FUNCTION

7. INSIDE A FLOW CYTOMETER
• 3 KEY SYSTEMS
• FLUIDICS
• OPTICS
• ELECTRONICS

8. What Happens in a Flow
Cytometer?
Cells in suspension flow single file
through a focused laser where they scatter
light and emit fluorescence that is filtered,
measured, then converted to digitized
values that are stored in a file
which can then be analyzed and interpreted
within specialized software.
Interrogation
Fluidics
Electronics
Interpretation

9. SIGNALS SEEN WITH FLOW
SCATTER
FORWARD SCATTER
(FSC) : PARALLEL OR
PERPENDICULAR
SIDE SCATTER (SSC)
FLUORESCENCE FITC,PE,APC,GFP,DAPI

11. FLUOROSCENT SIGNALS

12. BASIC USES OF FLOW
CYTOMETRY
PHENOTYPING
APOPTOSIS AND CELL DEATH
CELL CYCLE,DIVISION AND DNA SYNTHESIS
CELL TRACKING
SMALL PARTICLE ANALYSIS
FUNCTONAL ANALYSIS : CALCIUM FLUX,GENE
EXPRESSION,DYE EFFLUX
MICROORGANISM DETECTION

13. ADVANTAGES

14. DISADVANTAGES
COMMONLY
UNNECESSARY
EXPENSIVE TOO SLOW