Center for Advanced Light Microscopy (CALM)
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Point scanning confocal

SP5

Technique

The excitation beam path of a laser scanning confocal microscope focuses the excitation light to one diffratction limited spot in the specimen. Fluorescence light that is emitted from this spot is spatially filtered by a pinhole. In consequence, only light from a small 3D volume (voxel) that can be as small as 250nmx250nmx500nm (x,y,z) reaches the detector. By scanning this optical arrangement over the specimen, 2D optical sections can be aquired. Aquiring several 2D sections spaced in z allows to image 3D objects.

Use

Gold standard for imaging of optical sections, high-resolution 3D images and colocalisation studies. Living specimens often do not tolerate the high photon exposure. Reduced speed of image acquisition due to the point scanning principle.