Flow Cytometry- Introduction And Applications Flashcards
What is flow cytometry?
A technique that simultaneously measures several physical characteristics belonging to a single cell in suspension
What is the definition of flow cytometry?
Measuring the properties of cells in flow
What is flow sorting?
Sorting (separating) cells based on properties measured in flow
What is another name for flow sorting?
Fluorescence-activated cell sorting (FACS)
What can a flow cytometer tell us about a cell?
Relative size,
relative granularity/ internal complexity,
Relative fluorescence intensity
What can we measure with a flow cytometer?
Surface receptors
Intracellular cytokines/enzymes
Measure cell cycle viability and apoptosis
What are fluidics?
Cells in suspension flowing in a single file
How do fluidics work?
Inject a sample into a sheath fluid as it passes through a small hole
Simple fluid flows down a central core
What is hydrodynamic focusing?
Introduction of a large volume into a smaller volume
What are the steps in optical flow cytometry?
Illuminate fluid using a laser
Light scatters, emitting fluorescence
Fluorescence is collected and filtered
What is a laser line?
Single wavelength of light
What is the forward light scatter proportional to?
Size
What is a 90 degree light scatter proportional to?
Granularity
What can you do with a foward and 90degree light scatter?
Plot on a graph and identify the cell
How do you filter the light after scattering in optical cytometry?
Mirrors that separate out the light
What is the stokes shift?
The energy difference between the lowest energy peak of absorbance and the highest energy of emission
What are the names of the fluorochromes?
Fluorescein isothiocyanate (FITC)
Phycoerythin (PE)
Peridinin chlorophyll protein (PerCP)
What colour does FITC emit?
Green