Name: Leily Mahdavi

CORNELL NOTES
Class: Immunology Topic: Flow Cytometry
SHEET Date: 26/09/ 2021

QUESTIONS + Summary NOTES

SIZE, SHAPE, PROPERTIES Count and Analyze  SIZE, SHAPE, PROPERTIES of individual cells
Within a heterogenous population of cells

Quantitative data Flow cytometry software can analyze the data  QUANTITATIVE

Interrogation point
Forward scatter, Side scatter
Forward scatter directly
proportional to size
Side scatter is proportional to
shape and internal complexity
Procedure:
 Sample is extracted and placed in the flow cytometer
 Sample is mixed with saline solution in cytometer
 Cells passed through narrow one-cell-thick channel
 Cells fall down and pass through laser at interrogation point
 There is light scattered by each cell in two ways – Forward scatter and
Side scatter
 Forward scatter is detected by the laser and is proportional to size, the
Detector converts scatter into voltage pulse which is graphed by computer into
histogram
 Detector for scattered light is perpendicular to laser beam
 Side scatter is proportional to the shape
 Detector plots on histograms
 Therefore info about each cell’s Size, Shape, and Complexity
 This information can allow researchers to categorize the different cells
By their type (eg. Basophils)

Analysis of PopulationS++++ PRO: Can analyze multiple populations within sample

Detection of Fluorescent signal:

Fluorophores = fluorescent  The correct wavelength of laser light in the beam excites fluorophores
Molecules  Light emitted is passed through emission filters to allow for detection
Of multiple different fluorophores
 Can also quantify amounts of Flurophores
 Data presented through Histogram or dot plot

SUMMARY: Write 4 or more sentences describing specific learning from these notes.

Flow cytometry can tell you about the SIZE, SHAPE and INTERNAL COMPLEXITIES of cells as well as
giving information about Flurophores within cells.

Flow cytometry can analyze multiple cell types within one heterogenous sample.

Detection for; Forward scatter  Far side of laser beside cell. Side scatter  Perpendicular. Flurophores 
passed through emission filters._____