Micrometric size measurement of biological samples using a simple and non-invasive transmission interferometric set up.

Abstract

An interferometer with a minimum of optical hardware is employed to measure invasiveness the size of biological samples. Nowadays, there are several techniques in microscopy that render high quality resolved images. For instance, consider optical microscopy that has been around for over a century and has since developed in different configurations such as: bright and dark field, phase contrast, confocal, polarized, and so on. However, only a few of these use interferometry to retrieve not only the sample's amplitude but also its phase. An interesting example of the latter is digital holography which normally uses a Mach Zehnder interferometer setup. In the research work reported here a transmission digital holographic interferometer designed with a simple and minimal optical hardware, that avoids the drawback of the small field of view present in classical optical microscopic systems, is used to measure the microscopic dimensions of pollen grains. This optical configuration can be manipulated to magnify and project the image of a semitransparent sample over a neutral phase screen. The use of a collimated beam through the sample prevents geometrical distortions for high magnification values. The measurements using this novel configuration have been validated using a standard precision pattern displacement specimen with certified dimensions. As proof of principle, microscopically characterized pollen grains are placed in the transmission set up in order to estimate their dimensions from the interferometrically retrieved optical phase. Results match and thus show a relation between the sample's size and the optical phase magnitude.