Digital Image Files in Light Microscopy

Abstract

Publisher Summary The chapter discuses the image files from the standpoint of a non-computer scientist/graphics artist and emphasizes understanding the basic concepts and providing the individual biology researcher with the tools to make informed decisions about generating, manipulating, and storing graphics image files. Digital micrography can be used to represent single images of fixed material or to capture motion picture sequences of living cells. The use of computers for biological imaging follows an oral tradition based on colleagues and collaborators. While such an approach can propagate useful information, it can also perpetuate misconceptions, leading to bad digital habits. In many settings, standard protocols for generating and storing image files—on the basis of information from “the guy down the hall”—are developed without empirically determining which method is the best. With the rebirth of light microscopy as an analytical and quantitative tool, the use and manipulation of digital graphics files has become essential for the cell/ molecular biologist. Digital imaging has largely replaced 35-mm photography as a way to record micrographs. When used properly, digital image files allow for quantitative measurements of cellular activities. Proper use entails generating files with high bit depth (12 bit) and saving the files uncompressed. Images with lower bit depth (8 bit) can be used for presentation, and these files can be saved with compression to facilitate easy sharing.