65,000 Shades of Grey: Use of Digital Image Files in Light Microscopy

• Published in: Digital Microscopy Vol. 114; pp. 317 - 336
• Main Authors: Hinchcliffe, Edward, Shaw, Sidney
• Format: Reference Book Chapter Journal Article
• Language: English
• Published: United States Elsevier Science & Technology 2013
• Subjects: Animals; Cell Line; Data Compression; Humans; Image Processing, Computer-Assisted; Microscopy; Microscopy, Fluorescence - methods; Microscopy, Video - methods; Software industry; Codec; Compression; Image file; File format; Color
• ISBN: 0124077617; 9780124077614
• ISSN: 0091-679X
• DOI: 10.1016/B978-0-12-407761-4.00013-0

Computers dominate image capture and analysis in modern light microscopy. The output of an imaging experiment is a binary coded file, called an image file, which contains the spatial, temporal and intensity information present in the sample. Understanding what comprises an image file, and how these files are generated is necessary in order to optimize the use of the digital light microscope. In this chapter, we discuss image file formats, and the various components of these files, such as bit-depth, sampling rate, color theory, and compression, from the perspective of the non-computer scientist. We also discuss the problem of proprietary file formats, and how these often are incompatible with certain types of imaging software. We present several solutions to this issue. Finally, we present the use of digital movie formats, compression routines, and provide some real world examples for optimizing the generation of digital movies.