[48] Flow Cytometry

Question 1

What is flow cytometry?

Answer 1

A technique used to detect and measure physical and chemical characteristics of a population of cells or particles.

Question 2

What can flow cytometry measure?

Answer 2

• Size
• Complexity
• Relative granularity
• Fluorescence intensity

Question 3

What is a typical application of flow cytometry?

Answer 3

In immunology for counting and sorting cells, detecting biomarkers, and protein engineering.

Question 4

How does flow cytometry work?

Answer 4

Cells are suspended in a fluid stream, then passed individually through a laser beam; the fluorescence and light scatter is read by detectors.

Question 5

What are the components of a flow cytometer?

Answer 5

• Fluidics system
• Optics system
• Detection and analysis system

Question 6

What is a fluidics system in flow cytometry?

Answer 6

The part of the cytometer that transports the cells in a stream to the laser beam.

Question 7

What is the optics system in flow cytometry?

Answer 7

It comprises lasers that illuminate the cells and lenses that collect the light.

Question 8

What does the detection and analysis system in flow cytometry do?

Answer 8

It converts the detected light signals into electronic signals for data analysis.

Question 9

What are the requirements for a sample used in flow cytometry?

Answer 9

The sample must contain individual cells that can pass through the laser one at a time.

Question 10

Can flow cytometry measure more than one parameter at a time?

Answer 10

Yes, it can measure multiple parameters simultaneously by using different fluorescent dyes.

Question 11

What is a “gate” in flow cytometry?

Answer 11

A user-defined set of criteria that specify which events (cells) are included in further analysis.

Question 12

What is “FACS”?

Answer 12

Fluorescence Activated Cell Sorting, a specialized type of flow cytometry that sorts a heterogeneous mixture of cells into different containers.

Question 13

FACS - Initials

Answer 13

Fluorescence Activated Cell Sorting

Question 14

What is the advantage of FACS over standard flow cytometry?

Answer 14

FACS not only measures but also physically separates different cell populations.

Question 15

What are some limitations of flow cytometry?

Answer 15

• Cell characteristics must be measurable in terms of light scatter or fluorescence
• Cells must be in a single-cell suspension

Question 16

Can dead cells be identified in flow cytometry?

Answer 16

Yes, using specific dyes or antibodies that distinguish live and dead cells.

Question 17

What role does fluorescence play in flow cytometry?

Answer 17

Fluorescence is used to measure different parameters at the molecular level, such as protein expression.

Question 18

Can flow cytometry be used for DNA analysis?

Answer 18

Yes, it can measure DNA content and cell cycle status.

Question 19

How does flow cytometry contribute to cancer research?

Answer 19

It helps in phenotyping cells, determining cell health, and analyzing the cell cycle.