Laser Scanning Confocal Microscopy (LSCM/CLSM)
Confocal laser scanning microscopy (CLSM) or laser scanning confocal [LSCM]), often colloquially referred to simply as “confocal”, is a technique for obtaining high resolution optical images with depth selectivity. The key feature of confocal microscopy is its ability to acquire in-focus images from selected depths, a process known as optical sectioning. Images are acquired point-by-point and reconstructed with a computer. This allows three-dimensional reconstructions of topologically complex objects. However, CLSM is significantly slower than widefield or spinning-disk confocal microscopy, because images are acquired pixel-by-pixel and the laser power is concentrated on small diffraction limited spots which increases the phototoxicity load on the samples. CLSM typically requires high-laser power excitation, which can lead to phototoxicity and limited observation of live cells. This technology can be improved with image scanning mic/pixel reassignment to allow non-diffraction limited images which provide resolutions 100-200nm (check with the lab where these improvements are available).