Module Title – Quantitative Microscopy
Instructor – Sivaraj (Shiv) Sivaramakrishnan, Associate Professor, GCD
Description – This module will cover the basic workings of fluorescence microscopes and a sample quantitative analysis of image data. Part 1 of this module aims to introduce the incoming graduate student class, with a broad background in biological sciences to the essential workings of an epi-fluorescence microscope. Using fixed-cell samples stained with multiple fluorescent dyes students and working as pairs on a fluorescence microscope, we will systematically align and examine the workings of all the individual components including the light path, objective lenses, dichroics, camera, and software used for the acquisition of high quality fluorescence images of mammalian cells. Part 2 of this module will use FIJI (ImageJ) to analyze morphometric features in publication-quality image data to extract quantify information such as cell area, cell number, and also evaluate the image conditions vis-a-vis their suitability for quantitative analysis. Students will compare their analyzed data to published data in high-impact journals to better appreciate the rigor and caveats of appropriate imaging set up for quantitative data analysis.