Video Microscopy

Abstract

Video microscopy represents the first form of electronic imaging in light microscopy, and heralded the use of sophisticated fluorescence imaging techniques. However, today video microscopy – based on phase or differential interference optics – is considered to be an outdated technology. Rather than being yesterday's technology, a simple video microscope is a powerful tool for imaging cellular dynamics, and can easily be used to monitor cell cycle progression. Because the nonfluorescence imaging modes require significantly less light throughput, the video microscope is particularly suited for long‐term experiments, where photodamage can be a limiting factor. The video microscope is also an economical instrument that complements more sophisticated fluorescence‐based imaging systems, such as confocals, wide‐field deconvolution microscopes and super‐resolution microscopes. Here, I discuss a brief history of video microscopy, define some popular uses for the technology and discuss the selection of components for building a simple, yet functioning video microscopy system.

Key Concepts

  • Video microscopy represents the first electronic imaging system.
  • Simple video microscopes are an economical choice for live‐cell imaging.
  • System design requires selecting appropriate microscopes and illumination sources to allow long‐term imaging.
  • Live‐cell imaging requires control of temperature and pH.
  • Software selection to capture movies, control the imaging system and present the data is important.

Keywords: differential interference contrast; live cell imaging; mitosis; phase contrast; polarisation

Figure 1. Transmitted light contrast enhanced video microscopy. (a–a″). Differential interference contrast (DIC) microscopy. Frames from a time‐lapse sequence, showing the removal of the centrosome from a living cell by microsurgery. A microneedle was used to cut the cell into two fragments: a nucleus‐containing karyoplast and an enucleate cytoplast. Both fragments flatten out. Eventually, the karyoplast enters mitosis, as evidenced by it rounding up (see Hornick et al., ). The DIC reveals clear cell outlines, and the position of the nucleus. T = Hr:Min. (b). Phase contrast. A cluster of seven cells, with the central cell in mitosis (arrow). In these relatively thin specimens, this contrast mode clearly exposes the outlines of the individual cells, as well as the nucleus and dark nucleoli (see Hinchcliffe et al., ). Also, mitotic chromosomes are easily detected. (c). Polarisation microscopy. Sperm nuclei with associated centrosomes nucleate microtubule arrays when placed in Xenopus egg extracts. The position of each array – called an aster (like the flower) – appears as a black and white cross (see Hinchcliffe et al., ).
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Further Reading

Baird TR, Kaufman D and Brown CM (2014) Mercury free microscopy: an opportunity for core facility directors. Journal of Biomolecular Techniques 25: 48–53.

Goodwin P (2014) A primer on the fundamental principles of light microscopy: optimizing magnification, resolution, and contrast. Molecular Reproduction and Development. DOI: 10.1002/mrd.22385.

Khodjakov A and Rieder CL (2006) Imaging the division process in living tissue culture cells. Methods 38: 2–16.

Savoian MS (2015) Methods for live microscopy of drosophila spermatocytes. In: eLS. Chichester: John Wiley & Sons Ltd http://www.els.net 10.1002/9780470015902.a0020869.pub2.

Sluder G and Wolf DE (2013) Digital microscopy. Methods in Cell Biology 114. Elsevier Publishing, Amsterdam, NE. 672.

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Hinchcliffe, Edward H(Aug 2015) Video Microscopy. In: eLS. John Wiley & Sons Ltd, Chichester. http://www.els.net [doi: 10.1002/9780470015902.a0002638]