Cell Proliferation / Cytotoxicity Selection guide

Targets in Cell Proliferation, Viability, and Cytotoxicity

Various assays are used for assessing cell proliferation, viability, and cytotoxicity. Examples include optical counting, [3H] thymidine, MTT, WST, lactate dehydrogenase (LDH) assay, and more. This selection guide aims to provide you with information that will allow you to pick the method most suitable for your specific experimental needs.


Measure cell count and/or DNA content to reveal cell division and DNA replication, respectively.
– Optical counting with hemocytometer
– DNA quantification using fluorophores


Measure parameters of cellular health to determine the intracellular impact of cytotoxicity.
– Measuring evidence of cellular metabolic activity such as esterase activity, ATP, NADH dehydrogenase activity via MTT or WST, and so on


Measure indicators of cell death to evaluate cytotoxicity.
– Measure the activity of lactate dehydrogenase leaked out of damaged cell membranes
– Use membrane-impermeable dyes to selectively stain inside cells with damaged membranes

Seminar Video:
Reagent Selection Guide for Cell Proliferation/ Cell Viability / Cytotoxicity

0:00  Title
1:11  What are Cell Proliferation, Viability, and Cytotoxicity Tests?
4:12  Cell Viability: Comparison of Each Reagent and Assay Principle
8:53  Cytotoxicity: Comparison of Each Reagent and Assay Principle
13:52  Measurement of Multiple Parameters
16:33  More Detailed Cellular Function Analysis

Reagents Related to Cell Proliferation/Viability/Cytotoxicity Assays

The MTT and WST techniques are widely-used methods for evaluating cell proliferation and cytotoxicity. They are valued for the simplicity of their respective procedures, their safety, and the reproducibility of their results. WST (water-soluble tetrazolium) was developed by DOJINDO Laboratories in the 1990’s and is now used worldwide in cell proliferation assays. Cell Counting Kit-8 (CCK-8) is a commercial product that utilizes the WST method with improved sensitivity. Cell Counting Kit-8 is the original WST product, with which more than 5,000 papers have been published worldwide.

Product name MTT Cell Counting Kit-8 ATP Assay Kit Cell Counting Kit-F
Target Mitochondrial dehydrogenase activity Cellular dehydrogenase activity ATP Esterase activity
Instrument Microplate reader (colorimetric) Microplate reader (colorimetric) Microplate reader (luminometric) Microplate reader, FCM, Fluorescence microscope (fluorometric)
Advantage Low price · Simple procedure
· Highly stable reagent
· High sensitivity
· Short reaction time (15 min)
· Simple procedure
· Applicable for various instruments
· Short reaction time (15 min)
Disadvantage · Affected by reducing agents
· Requires dissolution of reagent after reaction
Affected by reducing agents · Expensive
· Half-life after light emission needs to be accounted for
· Serum increases background
· Leakage of reagent
Product code M009 CK04 CK06 (kit)
C396 (ready-to-use solution)
C326 (powder)


 Please watch this video for comparison between MTT and Cell Counting Kit-8.

Cell Proliferation Cytotoxicity Assay Kit_CCK-8

Cell Counting Kit-8 VS MTT

Comparison Among Cytotoxicity Assay Reagents

Product name Cytotoxicity LDH Assay Kit-WST Cell Counting Kit-F
Propidium iodide (PI) Trypan Blue
Target Released Lacate dehydrogenase (LDH) A marker (Calcein-AM) released into the supernatant (Calcein-Release Assay) Cytoplasm(Live cell membranes are impermeable to PI, but not dead cell membranes) Cytoplasm; cells taking up Trypan Blue are considered non-viable
Instrument Microplate Reader
Microplate reader, FCM, Fluorescence microscope
Fluorescence microscope, FCM Microscope
Advantage ·Simple procedure
·Suitable for multi-sample assay
·Convenient for cytotoxicity assay using 2+ kinds of cell samples ·Low price
·Simple procedure
·Low price
·No special instruments needed
Disadvantage ·Increased background by serum ·Leakage of reagent from living cells ·Not applicable for multi-sample assay ·False-positives due to cytotoxicity
·Not applicable for multi-sample assay
Product code CK12 CK06 (kit)
C396 (ready-to-use solution)
C326 (powder)
P378 (ready-to-use solution)
P346 (powder)

Cytotoxicity Assays Measuring Multiple Parameters

In order to verify the accuracy of results in a cytotoxicity assay, individual samples are measured according to multiple different principles. Cell Counting Kit-8 (code: CK04) and Cytotoxicity LDH Assay Kit-WST (code: CK12) measure dehydrogenase activity and the released amount of LDH as reflective of living and dead cells, respectively. As shown in the example below, separate measurements indicate that the number of living cells decrease, and the number of dead cells increase as a toxicant’s concentration increases.

Product Classification

Product Classification