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
Topic covered
Definition
Why
Methods
 The measure of proportion of alive and
metabolically active cell in a culture.
 Cell viability measurement based on
various cell function :
 Enzyme activity
 Cell membrane permeability
 Nucleotide uptake activity
 A colorimetric assay for the determination of
viable cell has been develop by using Tetrazolium
dye.
 Measure of cellular oxidative metabolism or
detect mitochondrial dehydrogenase activites in
the living cell
Tetrazolim dye MTT ( yellow colour )
NADPH –dependent oxidoreductase enz.
Formazon ( purple colour )
DMSO (Dimethyl sulfoxide) is added
It dissolve the insoluble formazon
Colored solution
O.D (500nm)
 The damaged cell membrane allows the spillage
of Lactate dehydrogenase into the medium.
The loss of cell viability is followed by an increase
in enzyme activity of lactate dehydrogenase
in the culture medium.
 They used as cell death marker ( eg adenylate
kinase and glucose-6-phosphate dehydrogenase)
 The intact cell growing in a culture media are
incubated with radioactivity labelled amino acids
or a nucleotide e.g Tritium –labelled thymidine
 The rate of protein synthesis or nucleic acid
synthesis can be measured by estimating
radioactivity of the culture .
• Trypan blue dye stained Dead cells not live cell .
As the dye penetrate the membrane of non –viable
cell which are stained blue .
The culture were microscopically monitored for
colony formation.
Very less cell density is used.