Multiplexing with multispectral imaging: From mice to microscopy

Richard M Levenson, David T. Lynch, Hisataka Kobayashi, Joseph M. Backer, Marina V. Backer

Research output: Contribution to journalArticle

61 Citations (Scopus)

Abstract

Increasing sophistication in the design and application of biological models as well as the advent of novel fluorescent probes have led to new demands on molecular imaging systems to deliver enhanced sensitivity, reliable quantitation, and the ability to resolve multiple simultaneous signals. Sensitivity is limited, especially in the visible spectral range, by the presence of ubiquitous autofluorescence signals (mostly arising from the skin and gut), which need to be separated from those of targeted fluorophores. Fluorescence-based imaging is also affected by absorbing and scattering properties of tissue in both the visible and to a lesser extent the near-infrared (NIR) regions. However, the small size of typical animal models (usually mice) often permits the detection of enough light arising even from relatively deep locations to allow the capture of signals with an acceptable signal-to-noise ratio. Multispectral imaging, through its ability to separate autofluorescence from label fluorescence, can increase sensitivity as much as 300 times compared to conventional approaches, and concomitantly improve quantitative accuracy. In the NIR region, autofluorescence, while still significant, poses less of a problem. However, the task of disentangling signals from multiple fluorophores remains. Multispectral imaging allows the separation of five or more fluorophores, with each signal quantitated and visualized separately. Preclinical small animal imaging is often accompanied by microscopic analysis, both before and after the in vivo phase. This can involve tissue culture manipulations and/or histological examination of fixed or frozen tissue. Due to the same advantages in sensitivity, quantitation, and multiplexing, microscopy-based multispectral techniques form an excellent complement to in vivo imaging.

Original languageEnglish (US)
Pages (from-to)78-88
Number of pages11
JournalILAR Journal
Volume49
Issue number1
StatePublished - 2008
Externally publishedYes

Fingerprint

Multiplexing
Microscopy
microscopy
Microscopic examination
image analysis
Fluorophores
Imaging techniques
fluorescent dyes
mice
Biological Models
Molecular Imaging
Optical Imaging
Signal-To-Noise Ratio
Fluorescent Dyes
Animals
Fluorescence
Tissue
Infrared radiation
Molecular imaging
Animal Models

Keywords

  • Autofluorescence
  • Fluorescence
  • In vivo imaging
  • Multiplexing
  • Reagents

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Levenson, R. M., Lynch, D. T., Kobayashi, H., Backer, J. M., & Backer, M. V. (2008). Multiplexing with multispectral imaging: From mice to microscopy. ILAR Journal, 49(1), 78-88.

Multiplexing with multispectral imaging : From mice to microscopy. / Levenson, Richard M; Lynch, David T.; Kobayashi, Hisataka; Backer, Joseph M.; Backer, Marina V.

In: ILAR Journal, Vol. 49, No. 1, 2008, p. 78-88.

Research output: Contribution to journalArticle

Levenson, RM, Lynch, DT, Kobayashi, H, Backer, JM & Backer, MV 2008, 'Multiplexing with multispectral imaging: From mice to microscopy', ILAR Journal, vol. 49, no. 1, pp. 78-88.
Levenson RM, Lynch DT, Kobayashi H, Backer JM, Backer MV. Multiplexing with multispectral imaging: From mice to microscopy. ILAR Journal. 2008;49(1):78-88.
Levenson, Richard M ; Lynch, David T. ; Kobayashi, Hisataka ; Backer, Joseph M. ; Backer, Marina V. / Multiplexing with multispectral imaging : From mice to microscopy. In: ILAR Journal. 2008 ; Vol. 49, No. 1. pp. 78-88.
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