Fluorescence Microscopy
Lasers are used in fluorescence microscopy to excite fluorophores and
reagents which are attached to the cell or organism under investigation. A specific area of fluorescence microscopy which utilises the high power available from lasers is called confocal microscopy. In confocal microscopy the laser light is scanned across the sample in a confocal configuration
and the fluorescence is detected through a confocal pinhole, which
gives very high resolution. Alternatively, the laser light passes
through rotating (spinning) disks of pinholes. In recent instruments the
laser can also be scanned across the sample as a line, which enables
very high frame rates, but requires higher power lasers. Confocal microscopy allows incredibly thin regions of cells or organisms to be easily and clearly visualised by discriminating out of focus light using pinholes. This microscopy method facilitates the investigation of cell dynamics, in particular of live cells, on a very high resolution scale.
Other applications related to this field are TIRF (total internal reflection fluorescence), FRAP (fluorescence recovery after photo bleaching), FCS (fluorescence correlation spectroscopy) and more recently techniques which allow observation of processes beyond the diffraction limit. These new super resolution techniques, SIM (structured illumination microscopy), PALM (photoactivated light microscopy), STORM (stochastic optical reconstruction microscopy) and STED (stimulated emission depletion microscopy) also require high power, high performance lasers.
The wavelength flexibility, low noise and high power make the Cobolt lasers highly suitable as excitation sources in most fluorescence microscopy applications.
See all Cobolt CW Visible 04-01 Series DPSS lasers and CW UV-Visible-NIR 05-01 Series DPSS lasers.
References
G.Karlsson et.al. Improved contrast with 473nm for confocal microscopy OSA CLEO CThII5 2003
C.Hertlein et.al. TIRF microscopy ACS 2007
L.Kaestner et.al. Novel 515 nm DPSS laser. Focus on Microscopy 2008
Photonik International Magazine March 2009 Fluorescence microscopy of living cells using new 515 nm DPSS lasers
S.Doose et. al. Single molecule FRET. Biophysics Letter 2007
M. Duser et. al Single molecule FRET SPIE Vol 6092 2006
N. Zarrabi et. al. Monitoring the rotary motors of single FF-ATP
Application notes
Cobolt Zouk™ for fluorescence microscopy
