Comparison of Different Methods to Measure Cell Viability

Cell viability assays are often used to screen collections of compounds to determine if the test molecules have effects on cell proliferation or show direct cytotoxic effects that eventually lead to cell death. According to the number and type of cells used, choosing a proper assay method decided whether the expected outcome is valid or not.

1. Comparison of Different Methods to Measure Cell Viability Creative Bioarray

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 · Creative Bioarray

3. · Creative Bioarray Methods The changes in cell physiology between the viable cell and dying cell are the base of cell viability assays. 04 Membrane Integrity 03 Mitochondrial Function 02 Apoptotic Markers 01 Cellular Metabolism

4. · Creative Bioarray 01 Low Repeatability of The Results 02 The Observed Results Differ From The Actual Measurements 03 The N umber or Type of Cells Both Can Influence The Detection R esult

5. · Creative Bioarray 01 03 02 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 Protease Activity A ssays The detections based on cell membrane integrity and related dye rejection have been widely accepted as a standard for cell viability testing. · Creative Bioarray

7. MTT MTS CCK-8 SRB

8. MTT Tetrazolium Reduction Assay

9. NADH ⇒ NAD + · Creative Bioarray

10. Direct correlation of formazan absorbance with B9 hybridoma cell number and time-dependent increase in absorbance. · Creative Bioarray

11. Disadvantages Advantages 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 may interfere with the optical readings. · Creative Bioarray

12. MTS Tetrazolium Assay

13. · Creative Bioarray 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.

14. · Creative Bioarray Simple Operation More S ensitive Good Repeatability

15. CCK-8 (WST-8) Assay

16. · Creative Bioarray Disengages The reagents are more expensive than MTT. It isn’t the most convenient method.

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

18. · Creative Bioarray Add CCK -8 solution Incubate for 1-4 hours Measure OD at 450nm

19. · Creative Bioarray 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

21. · Creative Bioarray 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 Measure 03

22. ·Creative Bioarray + + + + Procedure 02 Incubation of Cells With Treatment of Choice 01 Preparation of Treatment 03 Cell Fixation and SRB Staining 04 Absorbance Measurement 

23. · Creative Bioarray 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.

24. · Creative Bioarray 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, by Tris-base solution Convenience ++ +++ +++ + Detection speed + ++ +++ + Repeatability + ++ +++ ++ Stability + + ++ +++

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

27. · Creative Bioarray Contact Us Creative Bioarray www.creative-bioarray.com [email protected]