flow cytometry

Question 1

cytometry

Answer 1

technique for taking measurements on cells (usually done via microscopy)

Question 2

flow cytometry

Answer 2

measurements done while cells flow past detectors

Question 3

types of measurements

Answer 3

fluorescence + scatter

Question 4

main parts

Answer 4

fluidics + optics

Question 5

fluidics

Answer 5

hydrodynamic focusing → cells introduced to carrier fluid to align them to centre of the tube to be analysed

Question 6

light scattering

Answer 6

light passes through cells and is redirected + scattered into the detector by aspects of cell structure for view

Question 7

side scatter measurements

Answer 7

• provides measurements on internal structure / complexity of cell
• increase in side scatter is caused by granules + nucleus

Question 8

optical filters

Answer 8

filter between cells + detector → allows light of selected wavelengths to pass through while limiting others

Question 9

types of filters

Answer 9

standard band pass filter + dichroic filter

Question 10

flow imaging

Answer 10

records images of cells following process of flow cytometry (as opposed to raw data usually produced from flow cytometry)

Question 11

review of flow cytometry

Answer 11

• analyses single cells → detects diversity
• rapid + quantitative

Question 12

dye exclusion

Answer 12

• alive cells exclude dyes
• highlights dead cells as they can’t exclude it

Question 13

dye uptake

Answer 13

• alive cells allow permeation + dead block it
• highlights alive cells

Question 14

metabolic activity

Answer 14

• cells absorb + transform FDA (fluorescein diacetate) into fluorescent output
• shows which are performing metabolic activity

Question 15

fluorescence-activated cell sorting (FACS)

Answer 15

• cells picked for further studying are deflected by being given fluorescent dye
• charge is placed on them in order to be deflected from the chain of cells