cell death assay with fluorescent dyes + fluorescence microscopy + fluorophore + fluorescent dyes

dead cells can be differentiated from living cells by using

fluorescent dyes

when a cell dies, the plasma membrane disintegrates and allows

certain dyes to penetrate the cells

you can use the fluorescent dyes to

stain a cell culture and determine the ratio of dead vs living cells with a fluorescent microscope

fluorescence microscopes take advantage of

the difference in emission and transmission wavelengths of fluorophores to produce high contrast images

fluorescence microscopes produce

high contrast images

several fluorescent dyes

freely diffuse into a living cell

other fluorophores include fluorescent protein tags such as

a green fluorescent protein or snap tags

fluorescent protein tags can be expressed

transgenically and fluorescently labeled antibodies that bind to molecules of interest in the cell

fluorescent microscopy can be used to

visualize microscopic processes that are impossible to visualize with other microscopy techniques

when light is absorbed by a substance it results in

the excitement of an electron into a higher state

when the electron jumps back into the lower electronic state is

emits the energy as electromagnetic waves

fluorescence substances emit

electromagnetic waves within the visible light spectrum

what is the visible light spectrum

between 390 and 700 nanometers

fluorophores are used in techniques such as

fluorescent microscopy, qPCR, flow cytometry, and FACS

there are different ways of labelling a molecule fluorescently including

fluorescent dye

immunofluorescence

fluorescent tags

fluorescent dyes are

fluorophores that bind to specific cellular structures

the following fluorophores are frequently used as fluorescent dyes

DAPI

CY3 and CY5

Rhodamine

Fluorescein isothiocyanate

in many cases, there is

no fluorescent molecule that specifically binds to the cell structure to label

fluorescent molecules are linked to

another molecule that specifically binds to the cellular component to visualize