Low-cost, open-access refractive index mapping of algal cells using the transport of intensity equation

Abstract

Phase contrast microscopy allows stain free imaging of transparent biological samples. One technique, using the transport of intensity equation (TIE), can be performed without dedicated hardware by simply processing pairs of images taken at known spacings within the sample. The resulting TIE images are quantitative phase maps of unstained biological samples. Therefore, spatially resolved refractive index information can also be determined.\n\nUsing low-cost, open-source hardware, we applied the TIE to living algal cells to measure their refractive index. We obtained refractive index values that were repeatable within species and differed by distinct amounts depending on the species being measured. We suggest TIE imaging as a method of discrimination between different algal species and, potentially, non-biological materials, based on refractive index. Potential applications in biogeochemical modelling and climate sciences are suggested.