Methods

We provide infrastructure for the different techniques and methods listed below

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Confocal laser scanning microscopy (CLSM)

Confocal laser scanning microscopy (CLSM)

Confocal laser scanning microscopy has been established as a valuable tool for obtaining high resolution images and three-dimensional reconstructions of a variety of biological specimens.

The microscope works as follows

A beam of laser light is focused by an objective lens onto a fluorescent specimen. The fluorescent energy from the sample is then collected through the same objective and recorded by a photo detector. The optical system is designed so that the laser's focal point in the sample is imaged exactly on the face of the photo detector. This means that fluorescence coming from the point of laser focus will be focused on the photo detector, and fluorescence coming from any other points will be out of focus on the photo detector. By inserting a small aperture (pinhole) in front of the photo detector, the gathered fluorescence can be limited to a region very close to the focal point. In CLSM, the laser is moved across the sample in a raster (x-y) pattern, and by moving the focus vertically (z), multiple slices can be used to build up a full three-dimensional image.

Confocal laser scanning microscopy (CLSM)