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Photoactivation and optogenetics with micro mirror enhanced illumination

 
: Rückerl, Florian; Berndt, Dirk; Heber, Jörg; Shorte, Spencer

:

Thienpont, H. ; Society of Photo-Optical Instrumentation Engineers -SPIE-, Bellingham/Wash.:
Micro-Optics 2014 : Brussels, Belgium, April 2014
Bellingham, WA: SPIE, 2014 (Proceedings of SPIE 9130)
ISBN: 978-1-6284-1078-5
Paper 913017
Conference "Micro-Optics" <2014, Brussels>
English
Conference Paper
Fraunhofer IPMS ()
microscopy; diffractive MEMS; photoactivation

Abstract
Photoactivation and “optogenetics” require the precise control of the illumination path in optical microscopes. It is equally important to shape the illumination spatially as well as to have control over the intensity and the duration of the illumination. In order to achieve these goals we use programmable, diffractive Micro Mirror Arrays (MMA) as fast spatial light modulators for beam steering. By combining two MMAs with 256×256 mirrors each, our illumination setup allows for fast angular and spatial control at a wide spectral range (260-1000 nm). Illumination pulses can be as short as 50 µs, or can also extend to several seconds. The specific illumination modes of the individual areas results in a precise control over the light dose to the sample, giving significant advantage when investigating dosage dependent activation inasmuch as both the duration and the intensity can be controlled distinctly. The setup is integrated to a microscope and allows selective illumination of regions in the sample, enabling the precise, localized activation of fluorescent probes and the activation and deactivation of cellular and subcellular signaling cascades using photo activated ion channels. The high reflectivity in the UV range (up to 260nm) further allows gene silencing using UV activated constructs (e.g. caged morpholinos). Photoactivation and “optogenetics” require the precise control of the illumination path in optical microscopes. It is equally important to shape the illumination spatially as well as to have control over the intensity and the duration of the illumination. In order to achieve these goals we use programmable, diffractive Micro Mirror Arrays (MMA) as fast spatial light modulators for beam steering. By combining two MMAs with 256×256 mirrors each, our illumination setup allows for fast angular and spatial control at a wide spectral range (260-1000 nm). Illumination pulses can be as short as 50 µs, or can also extend to several seconds. The specific illumination modes of the individual areas results in a precise control over the light dose to the sample, giving significant advantage when investigating dosage dependent activation inasmuch as both the duration and the intensity can be controlled distinctly. The setup is integrated to a microscope and allows selective illumination of regions in the sample, enabling the precise, localized activation of fluorescent probes and the activation and deactivation of cellular and subcellular signaling cascades using photo activated ion channels. The high reflectivity in the UV range (up to 260nm) further allows gene silencing using UV activated constructs (e.g. caged morpholinos).

: http://publica.fraunhofer.de/documents/N-294224.html