Hostname: page-component-78c5997874-lj6df Total loading time: 0 Render date: 2024-11-16T13:25:33.602Z Has data issue: false hasContentIssue false
  • English
  • Français

Light-Emitting Diodes Are Better Illumination Sources for Biological Microscopy than Conventional Sources

Published online by Cambridge University Press:  03 March 2008

Richard W. Cole  and
James N. Turner
Richard W. Cole*
Affiliation:
Wadsworth Center, New York State Department of Health, P.O. Box 509, Albany, NY 12201, USA
James N. Turner
Affiliation:
Wadsworth Center, New York State Department of Health, P.O. Box 509, Albany, NY 12201, USA
*
Corresponding author. E-mail: [email protected]
Get access

Abstract

Light-emitting diodes (LEDs) can be easily and inexpensively integrated into modern light microscopes. There are numerous advantages of LEDs as illumination sources; most notably, they provide brightness and spectral control. We demonstrate that for transmitted light imaging, an LED can replace the traditional tungsten filament bulb while offering longer life; no color temperature change with intensity change; reduced emission in the infrared region, which is important for live cell imaging; and reduced cost of ownership. We show a direct substitution of the typical tungsten bulb with a commercially available LED and demonstrated the color stability by imaging a histology section over a wide range of light intensities. For fluorescent imaging, where the typical illumination sources are mercury or xenon lamps, we demonstrate that LEDs offer advantages of providing a longer lifespan, having a more constant intensity output over time, more homogeneous illumination, and significantly lower photon dose. Our LED equipped system was used to image and deconvolve dual fluorescently labeled cells, as well as image cells undergoing mitosis expressing green fluorescent protein–histone 2B complex. The timing of the stages of mitosis is well established as an indicator of cell viability.

Keywords

time-lapse microscopylight-emitting diodes (LEDs)green fluorescent protein (GFP)absorbent stainsfluorescence imagingdeconvolution
Type
Biological Applications
Information
Microscopy and Microanalysis , Volume 14 , Issue 3 , June 2008 , pp. 243 - 250
Copyright
Copyright © Microscopy Society of America 2008

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

REFERENCES

Benavides, J.M. & Webb, R.M. (2005). Optical characterization of ultrabright LEDs. Appl Optics 44, 4000–4003.Google Scholar
Bormuth, V., Howard, J. & Schaffer, E. (2007). LED illumination for video-enhanced DIC imaging of single microtubules. J Microsc 226, 1–5.Google Scholar
Eltoum, I.A. & Roberson, J. (2007). Impact of HPV testing, HPV vaccine development, and changing screening frequency on national Pap test volume: Projections from the National Health Interview Survey. Cancer 111, 34–40.CrossRefGoogle ScholarPubMed
Hogan, H. (2007). LEDs have a bright future in biomedical instruments. Biophotonics Int 3, 36–39.Google Scholar
Moser, C., Mayr, T., Klimant, L., Morgan, C.G., Mitchell, A.C., Heinlein, T., Biebricher, A., Schluter, P., Roth, C.M., Herten, D.P., Wolfrum, J., Heilemann, M., Muller, C., Tinnefeld, P. & Sauer, M. (2006). Filter cubes with built-in ultrabright light-emitting diodes as exchangeable excitation light sources in fluorescence microscopy. J Microsc 222, 135–140.CrossRefGoogle ScholarPubMed
Nishigaki, T., Wood, D.C., Shiba, K., Baba, S.A. & Darszon, A. (2006). Stroboscopic illumination using light-emitting diodes reduces phototoxicity in fluorescence cell imaging. Biotech 41, 191–197.CrossRefGoogle ScholarPubMed
Rieder, C.L. & Cole, R.W. (1998). Entry into mitosis in vertebrate somatic cells is guarded by a chromosome damage checkpoint that reverses the cell cycle when triggered during early but not late prophase. J Cell Bio 142, 1013–1022.Google Scholar
Wallace, W., Schaefer, L.W. & Swedlow, J.R. (2001). A workingperson's guide to deconvolution in light microscopy. Biotech 31, 1076–1078.Google Scholar