Abstract

We report on the application of a novel all-solid-state violet laser diode source to confocal microscopy. The source has the potential to replace argon ion lasers in a range of fluorescence based imaging systems. Improvements in system performance and image quality through the use of anamorphic prisms to modify the beam profile have been characterised. These modifications have permitted high quality, optically sectioned images to be obtained from laser diodes operating around 406nm. Living mammalian cells stained with a range of biologically significant fluorophores have been imaged. In addition, it has been shown that at this wavelength it is possible to image dyes that normally require excitation with UV argon laser lines.

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References

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    [CrossRef] [PubMed]

Other

J. B. Pawley (Ed.), "Handbook of Biological Confocal Microscopy,"2 nd Edition, Plenum (1995).

R. P. Haugland, "Handbook of Fluorescent Probes and Research Chemicals, 6th Edition," Molecular Probes Inc.

S. Nakamura, M. Senoh, S. Nagahama, N. Iwasa T. Yamada, T. Matsushita Y. Sugimoto H. Kiyoku, "Optical gain and carrier lifetime of InGaN multi-quantum well structure laser diodes," Appl. Phys Lett 69 1568-1572 (1996).
[CrossRef]

S. Nakamura "Developments in blue and UV optical semiconductor sources," Proc. SPIE 3621-01 125-135 (1999).

J. Bewersdorf, S. W. Hell, "Picosecond pulsed two-photon imaging with repetition rates of 200 and 400 MHz," Journal of Microscopy 191 28-38 (1998).
[CrossRef]

P. E. Hockerber, T. A. Skimina, V. E. Centonze, C. Lavin, S. Chu, S. Dadras, J. K. Reddy, J. G. White, "Activation of flavin-containing oxidases underlies light-induced production of H2O2 in mammalian cells," Proc. Natl. Sci. 96 6255-6260 (1999).
[CrossRef]

A. Miyawaki, J. Llopis, R. Heim, J.M. McCaffery, J.A. Adams, M. Ikura, R. Tsien, "Fluorescent indicators for Ca 2+ based on green fluorescent protein and calmodulin," Nature 388 882-887 (1997).
[CrossRef] [PubMed]

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Figures (2)

Figure 1.
Figure 1.

Axial profile x63 1.35NA objective, laser diode with and without anamorphic beam shaping

Figure 2.
Figure 2.

Live vascular smooth muscle cell stained with FITC imaged directly, 2a directly with laser diode, 2b laser diode with anamorphic beam shaping, 2c Cos-7 cell with eGFP transfection

Tables (2)

Tables Icon

Table 1. Measured optical performance

Tables Icon

Table 2. Summary of flurophores imaged with 406nm

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