The Leica TCS SP2 is a spectral confocal with 3CH+DIC digital emission 400-800nm range designed to acquire high spatial resolution images of fluorescently labeled materials and for analysis of these images. The confocal principle utilizes a pinhole (confocal aperture) to eliminate out-of-focus light from fluorescently labeled specimens. Lasers provide intense, point illumination that is scanned over the preparation and the fluorescence at each point is quantified and used to construct a representation of the object brightness (an image). This method provides high resolution for the x and y planes as well as vertically (z plane).
Our SP2 confocal system has seven laser excitation lines: 458, 476, 488, 496, 514, 543, and 633, all fiber coupled and controlled by AOTFs. It has a laser scanner with up to 2000-Hz line scans, frame scan rate of 40 fps at 512x32 pixels, scan resolution up to 4096x4096 pixels, zoom to 32x, scan rotation; interactive control panel with digital potentiometers. The scan detector is equipped with an acousto-optical beam splitter (AOBS) to select/introduce most excitation laser lines, eliminating the need for main dichroic mirrors. The system has three PMT detectors with 8 or 12-bit output, simultaneous operation and digital/graphic definition of emission wavelength content per channel. Spectral scanning / display and linear un-mixing are possible, giving the ability to separate largely overlapping, similar colored fluorophores (e.g., GFP vs. FITC or YFP).
LCS Leica Confocal Software includes LCS 3D Volume rendering, 3D animations, 3D filter, Stereo images and animations, Average projection, Maximum point projection, SFP; LCS Multicolor Co-localization of 2channel and 3channel recordings. Cytofluorograms in 2D and 3D display. Histogram-segmentation and masking.Use of the confocal system requires in-person training to make sure the system remains available to Users.