The document compares various methods of measuring cell viability, such as MTT, CCK-8, and SRB assays, highlighting their advantages and disadvantages. It details the principles of these assays, including their detection methods, solubility, and operational complexities. Creative Bioarray offers these assays for applications in drug screening and cytotoxicity testing.

1. Comparison of Different
Methods to Measure Cell
Viability
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2. Cell Viability
Cell viability is a determination
of living or dead cells, based on
a total cell sample.
01
Drug
Screening
02
Cytotoxicity
Tests of
Chemicals
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Methods
The changes in cell physiology between the viable cell and dying cell are the
base of cell viability assays.
04Membrane
Integrity
03Mitochondrial
Function02Apoptotic
Markers
01Cellular
Metabolism

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01
Low Repeatability
ofThe Results
02
The Observed Results
Differ FromThe Actual
Measurements 03
The Number or Type of
Cells Both Can Influence
The Detection Result

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01 0302 04
The product is colored or
fluorescent that can be
detected with a plate reader
But the dying cells lose
the conversion ability
Viable cells can convert
the substrate into
product
The signal is directly
proportional to the number of
viable cells present

6. Tetrazolium
Reduction
Resazurin
reduction
ProteaseActivity
Assays
The detections based on cell membrane
integrity and related dye rejection have
been widely accepted as a standard for
cell viability testing.
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7. MTT MTS CCK-8 SRB

8. MTT Tetrazolium
Reduction Assay

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10. Direct correlation of formazan absorbance with B9 hybridoma
cell number and time-dependent increase in absorbance.
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11. DisadvantagesAdvantages
• Fewer steps, uses
fewer materials, and
does not carry the
added burden of
radioactive waste
disposal.
• Not suitable for
suspending cells.
• Must be optimized for
seeding amount and assay
duration to obtain
satisfactory results.
• Precipitated protein and
cellular debris present in
the culture plate wells
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12. MTS Tetrazolium
Assay

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Approach
For these improved
tetrazolium reagents, the
usual protocol is simply to
add them to the culture
medium and read the
absorption of the reduced
product at an appropriate
time.

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Simple
Operation
More Sensitive
Good Repeatability

15. CCK-8 (WST-8)
Assay

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Disengages
• The reagents are more expensive
than MTT.
• It isn’t the most convenient method.

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Principle
A: Structures of WST-8
and WST-8 formazan.
B: Principle of the cell
viability detection with
Cell Counting Kit-8.

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Add CCK -8 solution Incubate for 1-4 hours Measure OD at 450nm

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01
One-bottle solution; no premixing of components is required
02
Simple, Rapid and Low Consumed
03
Good repeatability and Nonradioactive

20. Sulforhodamine B
(SRB) colorimetric
assay

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03 Measure
As the binding of SRB is stoichiometric, the
amount of dye extracted from stained cells can
be used as a proxy for cell mass directly, and the
amount of bound dye is proportional to the cell
mass.
01 Binding
SBR bind to basic amino-acid
residues components of cells under
mildly acidic conditions.
02 Dissociation
The binding will dissociate under basic
conditions.Thus, the amount of bound
dye can be used as a proxy for cell
mass, which can then be extrapolated
to measure cell proliferation.
Dissociation
02
Binding
01
M
easure
03

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+ ++ +
Procedure
02Incubation of
Cells With
Treatment of
Choice
01
Preparation
of
Treatment
03
Cell Fixation
and SRB
Staining
04
Absorbance
Measuremen
t

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Advantages
• SRB staining rarely affected by other compounds interfere which can
remain for a long time.
• The 96-hole cell culture plate at different time points can be
determined at the same time.

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• The application of the SRB assay is limited
to manual or semiautomatic screening due
to the multiple washing and drying steps,
which are not amenable to automation.
• The operation procedure is complicated.
Disadvantages

25. · Summary
MTT MTS CCK-8 SRB
Solubility Indissolvable Water soluble Water soluble Indissolvable
Detection Wavelength 490nm 450nm 450nm 510nm
Character Solid Liquid Liquid Liquid
Usage
Prepare the
solutions
Use it right after it was
ready
No need to prepare Prepared beforehand
Need to redissolve or not Yes, by DMSO No No
Yes, byTris-base
solution
Convenience ++ +++ +++ +
Detection speed + ++ +++ +
Repeatability + ++ +++ ++
Stability + + ++ +++

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Cell
Viability
BrdUMTS
MTT
CCK-8
SBR
Services & Products
Creative Bioarray offers a wide selection of cell viability assays for
drug screening and cytotoxicity tests of chemicals.

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Contact Us
Creative Bioarray
www.creative-bioarray.com
info@creative-bioarray.com