Microscopy Facility
Confocal Laser Scanning Microscopy
Confocal laser scanning microscopy allows for high resolution, multi-channel three-dimensional imaging of fluorescently-labeled or reflective specimens. The advantage over conventional widefield light microscopy is that the optics of the confocal microscope remove “blurred” light originating from outside the focal plane of interest, thus generating an “optical section.” Three-dimensional image reconstruction of serial optical sections as well as quantitative measurements may be performed using the microscope software.
The facility houses a Leica TCS-SP2 AOBS confocal laser scanning microscope (inverted) with a spectrophotometer scan head, a high resolution Märzhäuser MCX-2 motorized XY stage and three confocal detectors (plus a transmitted light detector). The system has five lasers: blue diode (405 nm), Argon (458, 476, 488, 514 nm), green HeNe (543 nm), orange HeNe (594 nm) and red HeNe (633 nm). The spectrophotometer scan head allows the user to “tune” the detectors to any emission wavelength.
In addition to being able to tune to different emission wavelengths, the SP detector allows for spectral scanning of fluorescence and emission “fingerprinting” as well as linear unmixing of signals from fluors with overlapping emission profiles. The system is also capable of FRET (fluorescence resonance energy transfer) and FRAP (fluorescence recovery after photobleaching) imaging.
A PeCon stage incubator is available to permit live cell confocal imaging.
