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Table 1 A summary of the advantages and disadvantages of the four cell enumeration assays

From: Limitations of the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay when compared to three commonly used cell enumeration assays

 

Advantages

Disadvantages

Neutral red uptake assay

1. Cell enumeration independent of enzymatic conversion of dye [30,31]

1. Some reports of test compound interference [32]

2. Few wash steps involved [30,31]

MTT assay

1. Gold standard for cytotoxicity testing

1. Conversion to formazan crystals depends on metabolic rate and number of mitochondria resulting in many known interferences [4,12-17,33]

2. Suitable for high-throughput screening and miniaturisation [34]

2. Numerous wash steps required [3,7]

Resazurin reduction assay

1. Few wash steps involved [19]

1. Conversion to resorufin depend on enzymatic conversion [18]

2. Follow-up assays can be performed on same cells as assay is not cytotoxic [32,35]

2. Absorbance-based method less sensitive than fluorescence-based method

Sulforhodamine B assay

1. Cell enumeration dependent on protein content thus no test compound interference [21,22]

1. Numerous wash steps involved, but fixation required [22]

2. Highly reproducible

2. Less sensitive with non-adherent cells