Electronic cameras for low-light microscopy

Methods Cell Biol. 2013:114:211-41. doi: 10.1016/B978-0-12-407761-4.00010-5.

Abstract

This chapter introduces to electronic cameras, discusses the various parameters considered for evaluating their performance, and describes some of the key features of different camera formats. The chapter also presents the basic understanding of functioning of the electronic cameras and how these properties can be exploited to optimize image quality under low-light conditions. Although there are many types of cameras available for microscopy, the most reliable type is the charge-coupled device (CCD) camera, which remains preferred for high-performance systems. If time resolution and frame rate are of no concern, slow-scan CCDs certainly offer the best available performance, both in terms of the signal-to-noise ratio and their spatial resolution. Slow-scan cameras are thus the first choice for experiments using fixed specimens such as measurements using immune fluorescence and fluorescence in situ hybridization. However, if video rate imaging is required, one need not evaluate slow-scan CCD cameras. A very basic video CCD may suffice if samples are heavily labeled or are not perturbed by high intensity illumination. When video rate imaging is required for very dim specimens, the electron multiplying CCD camera is probably the most appropriate at this technological stage. Intensified CCDs provide a unique tool for applications in which high-speed gating is required. The variable integration time video cameras are very attractive options if one needs to acquire images at video rate acquisition, as well as with longer integration times for less bright samples. This flexibility can facilitate many diverse applications with highly varied light levels.

Keywords: Complementary metal-oxide semiconductor; Frame rate; Gain; Low-Light Imaging; Quantum efficiency.

MeSH terms

  • Cells, Cultured
  • Humans
  • Microscopy, Fluorescence / instrumentation
  • Microscopy, Fluorescence / methods
  • Microscopy, Video / instrumentation
  • Microscopy, Video / methods
  • Photography / instrumentation
  • Photography / methods*
  • Signal-To-Noise Ratio
  • Video Recording / instrumentation
  • Video Recording / methods*