Microscale Colorimetric Analysis Using a Desktop Scanner and Automated Digital Image Analysis

In recent decades, several standard colorimetric reactions for chemical analysis have been miniaturized to microwells on microplates, including methods useful for environmental measurements. Advantages of method miniaturization include a reduction in reagents required, improved safety, reduced waste stream, and increased sample throughput. However, the widespread use of microscale techniques employing microplates in classroom settings is likely limited by the high cost of microplate readers. Although spectrophotometers read peaks of specific wavelengths, absorbance spectra tend to be relatively broad and measurements at specific wavelengths are highly autocorrelated with those of nearby neighbors, which implies that broadband intensity data of red, green, and blue channels may indeed be adequate for digital colorimetric quantification. In this article, we demonstrate that digital image analysis of a scanned microplate image can substitute for a spectrophotometer for several common quantitative microscale procedures. This finding allows for cost effective and microscale quantification of several compounds to be demonstrated in the laboratory. Additionally, popular teaching and learning activities such as water quality monitoring can now be performed accurately and inexpensively using digital image analysis.