Evaluation of redox indicators and the use of digital scanners and spectrophotometer for quantification of microbial growth in microplates

J Microbiol Methods. 2002 Jun;50(1):63-73. doi: 10.1016/s0167-7012(02)00011-8.

Abstract

The growth indicators 2,3,5-triphenyltetrazolium chloride (TTC), 2-[4-iodophenyl]-3-[4-dinitrophenyl]-5-phenyltetrazolium chloride (INT), 2,3-bis[2-methoxy-4-nitro-5-sulfophenyl]-2H-tetrazolium-5-carboxanilide inner salt (XTT), 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT), and resazurin were tested for their ability to indicate bacterial growth/growth inhibition. Two reading devices were evaluated and compared, a microplate spectrophotometer and a digital flatbed scanner. The bacteria used in the study were cultivated in 96-wells microplates and readings were made after 24 h. The scanned pictures were analysed with a software developed in-house to generate numerical values. It was found that resazurin was difficult to use since it shifts between three colours. MTT and TTC had a high correlation between the spectrophotometer data and the data from the scanned images. The reproducibility was similar for both reading devices. In no case was there a need to resuspend the pellets before reading. Both the XTT and INT showed lower correlations. It is concluded that bacterial growth/growth inhibition can be easily and reproducibly measured from microplate cultivations with a flatbed scanner or with a microplate spectrophotometer.

Publication types

  • Comparative Study
  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacteria / growth & development*
  • Culture Media
  • Indicators and Reagents / chemistry*
  • Microbiological Techniques
  • Oxidation-Reduction
  • Signal Processing, Computer-Assisted*
  • Spectrophotometry / instrumentation
  • Spectrophotometry / methods*
  • Tetrazolium Salts / chemistry*

Substances

  • Culture Media
  • Indicators and Reagents
  • Tetrazolium Salts