Flow cytometry

Fluorescent?

Fluorescent?

Signal is detected in same direction as side scatter and directed through optical filters to chose range to deliver to detector.

Define the speed of flow cytometer

It is relatively fast with an acquisition rate of 10000 events per second.

Fluidics?

aligns cells by hydrodynamic focusing to present at interrogation point.

Optics

consist of light sources to illuminate sample.

Fluorescence emission

________: informs about specific parameters, fluorophore specifically bound to cells and cellular components.

Use of flow cytometer in HIV screening

It can be used in HIV screening because there is a decline in T- helper lymphocytes so we need to count the number of T- helper cells during screening.

Drop formation

________: the flow head is shaken by a piezoelectric vibrator (30- 40 kHz) to decompose the fluid that leaves the head into droplets (1 droplet= 1 cell)

Fluidics

aligns cells by hydrodynamic focusing to present at interrogation point

Optics

consist of light sources to illuminate sample

Electronics, data analysis

convert light from detectors to digital data

Sorting

physically separates interesting subsets of cells based on parameters of others

Light scattering can be?

Forward scattering Or Side scattering

Fluorescence emission

informs about specific parameters, fluorophore specifically bound to cells and cellular components

List mode

presentation of measured data in all cells, raw data stored on computer

Single parametric representation(Histogram)

plot of number cells characterised by a given parameter

Multiparametric representation(Dot plot)?

applies the correlation method, corresponding values of two measured parameters for each cell (x, y axis) are displayed as dots, each dot corresponds to a single particle

Gating?

definition of subpopulation of cells in one-, or multiparametric representation for detailed investigation or separation

Drop formation

the flow head is shaken by a piezoelectric vibrator (30-40 kHz) to decompose the fluid that leaves the head into droplets (1 droplet = 1 cell)

Electrostatic separation

the droplet to be separated is charged at the place of droplet formation and deflected by an electric field

Flow cytometry?

Flow cytometry allows that in a flowing heterogeneous suspension of cells, individual cells and their characteristics can be measured. On the basis of measured parameters cells subsets can be separated for further purposes.

Application of flow cytometry

Medical biology and clinical practice

Cell separation

physical separation of cell populations characterized by a given parameter (FSC, SSC, FL) for further

investigation (Biochemical analysis. cell culturing).