What is the principle of light sheet microscopy?
optical sectioning by illuminating specimen along optical path orthogonal to detection axis
illumination of only thin layer -> better focus and minimized photobleaching
What kind of lense is used for light sheet microscopy?
cylindrical lense combined with Gaussian beam or axicon lense combined with bessel beam -> focus light beam along line, not a single point
What are different setups of lense and beam for light sheet microscopy?
Selective plane illumination microscopy (SPIM): laser -> cylindrical lense -> light sheet
Down-scanned light sheet microscopy (DSLM): rapidly up and down scanning laser → virtual light sheet
1-photon Bessel beam: DSLM with Bessel beam -> even thickness
2-photon Bessel beam: DSLM with pulsed IR laser → two-photon excitation → highest intensity in central core of the beam
Compare Gaussian and Bessel beam
What type of detection/illumination configurations are there for light sheet microscopy?
inverted configuration 1-view
inverted configuration 2-view
upright configuration 1-view
MultiView
What is the improvement of lattice light sheet microscopy?
lowers the spread due to refraction
creates 2D lattice of regularly spaced Bessel beams -> increased axial resolution
but many unwanted frequencies, that have to be filtered out
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