Improving fluorescence confocal microscopy with cryogenically-cooled diode lasers

doi:10.1364/OPEX.12.004157

Optics Express

Editor: Michael Duncan
Vol. 12, Iss. 17 — Aug. 23, 2004
pp: 4157–4165

Improving fluorescence confocal microscopy with cryogenically-cooled diode lasers

Nelson Sivers, Brian Van de Workeen, Susanne Lee, James Turner, and Donald Szarowski »View Author Affiliations

Optics Express, Vol. 12, Issue 17, pp. 4157-4165 (2004)
http://dx.doi.org/10.1364/OPEX.12.004157

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Abstract
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Abstract

Recent advances in fluorescence confocal microscopy have focused on exciting multiple dyes, leading to the use of diode laser sources. We show that by varying a diode’s operating parameters, diode lasers can address some of the inherent problems associated with multiple dye excitation. Cooling the laser diode caused its emission wavelengths to decrease linearly, its output power to increase five times, and the noise due to reflected laser light for equal fluorescence signals to be reduced five times. We utilized these improvements to produce fluorescence confocal images that minimized the reflected laser intensity, while still efficiently exciting the fluorochrome.

OCIS Codes
(140.2020) Lasers and laser optics : Diode lasers
(180.1790) Microscopy : Confocal microscopy

ToC Category:
Research Papers

History
Original Manuscript: June 21, 2004
Revised Manuscript: August 16, 2004
Published: August 23, 2004

Citation
Nelson Sivers, Brian Van de Workeen, Susanne Lee, James Turner, and Donald Szarowski, "Improving fluorescence confocal microscopy with cryogenically-cooled diode lasers," Opt. Express 12, 4157-4165 (2004)
http://www.opticsinfobase.org/oe/abstract.cfm?URI=oe-12-17-4157

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References

J. M. Girkin; A. I. Ferguson; D. L. Wokosin; and A. M. Gurney, �??Confocal microscopy using and InGaN violet laser diode at 406nm�??, Opt. Express 7, 336-41 (2000), <a href="http://www.opticsexpress.org/abstract.cfm?URI=OPEX-7-10-336">http://www.opticsexpress.org/abstract.cfm?URI=OPEX-7-10-336</a> [CrossRef] [PubMed]
See for example Bio-Rad�??s Radiance2100�?�, <a href="http://cellscience.bio-rad.com/products/confocal/Radiance2100/default.htm">http://cellscience.bio-rad.com/products/confocal/Radiance2100/default.htm</a>
See for example Coherent Inc.�??s data sheet <a href="http://www.coherent.com/Downloads/Radius635_DSandHeNeCC_100103.pdf.">http://www.coherent.com/Downloads/Radius635_DSandHeNeCC_100103.pdf</a>.
P. M. Conn, Methods in Enzymology Volume 307 Confocal Microscopy (Academic Press, San Diego, 1999). [CrossRef]
C. J. R. Sheppard and D. M. Shotton, Confocal Laser Scanning Microscopy (Springer-Verlag, New York, 1997).
C. J. Hawthorn, K. P. Weber, and R. E. Scholten, �??Littrow configuration tunable external cavity diode laser with fixed direction output beam�??, Rev. Sci. Instrum 72, 4477-4497 (2001). [CrossRef]
S.M. Sze, Physics of Semiconductor Devices (John Wiley & Sons, New York, 1981), p. 264.
S.M. Sze, Physics of Semiconductor Devices (John Wiley & Sons, New York, 1981), p. 91-92.
Bio-Rad Fluorochrome database, <a href="http://cellscience.bio-rad.com/fluorescence/fluorophoreDatab.htm">http://cellscience.bio-rad.com/fluorescence/fluorophoreDatab.htm</a>

Methods in Enzymology

P. M. Conn, Methods in Enzymology Volume 307 Confocal Microscopy (Academic Press, San Diego, 1999). [CrossRef]

Opt. Express

J. M. Girkin; A. I. Ferguson; D. L. Wokosin; and A. M. Gurney, �??Confocal microscopy using and InGaN violet laser diode at 406nm�??, Opt. Express 7, 336-41 (2000), <a href="http://www.opticsexpress.org/abstract.cfm?URI=OPEX-7-10-336">http://www.opticsexpress.org/abstract.cfm?URI=OPEX-7-10-336</a> [CrossRef] [PubMed]

Rev. Sci. Instrum

C. J. Hawthorn, K. P. Weber, and R. E. Scholten, �??Littrow configuration tunable external cavity diode laser with fixed direction output beam�??, Rev. Sci. Instrum 72, 4477-4497 (2001). [CrossRef]

Other

S.M. Sze, Physics of Semiconductor Devices (John Wiley & Sons, New York, 1981), p. 264.
S.M. Sze, Physics of Semiconductor Devices (John Wiley & Sons, New York, 1981), p. 91-92.
Bio-Rad Fluorochrome database, <a href="http://cellscience.bio-rad.com/fluorescence/fluorophoreDatab.htm">http://cellscience.bio-rad.com/fluorescence/fluorophoreDatab.htm</a>
See for example Bio-Rad�??s Radiance2100�?�, <a href="http://cellscience.bio-rad.com/products/confocal/Radiance2100/default.htm">http://cellscience.bio-rad.com/products/confocal/Radiance2100/default.htm</a>
See for example Coherent Inc.�??s data sheet <a href="http://www.coherent.com/Downloads/Radius635_DSandHeNeCC_100103.pdf.">http://www.coherent.com/Downloads/Radius635_DSandHeNeCC_100103.pdf</a>.
C. J. R. Sheppard and D. M. Shotton, Confocal Laser Scanning Microscopy (Springer-Verlag, New York, 1997).

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