User manual LEICA TCS SP5 TANDEM SCANNER BROCHURE

[. . . ] Regarding confocal microscopy, the systems on the market, today, are either optimized for morphological image acquisition ­ where sufficient time is available for image restoration ­ or for rapid data acquisition ­ where speed is the main concern whereas beauty is not an issue. In the latter case the recording parameters are mostly set in a way to , , just see something behind the noise". This means one has to invest twice, if research shall be done in both categories. [. . . ] Ca2+ waves in a living sample a conventional scanning system is far too slow. But , , tuning" it to higher speeds brings significant limitations (field of view). Here, a new solution by Leica Microsystems solves the problem: Resonant Scanner The resonant scanning system of the Leica TCS SP5 works at 8000 Hz frequency and enables to record data in both directions of the scan motion. It thus acquires about 25 images per second at a frame resolution of 512 x 512 pixels. This is about 10 times faster compared to the recording speed of conventional scanners. For even faster events, one can reduce the number of lines per image managing up to 200 frames per second. For extremely fast changes, data can be recorded by images containing only one single line. Large field of view The application of resonant scanners has additional advances compared to other high-speed solutions. The field of view is very large (15 mm fov) compared to conventional scanners, where the field of view at 2000 Hz is restricted to some 3 mm fov. This does not work for systems that employ cameras (typically spinning-disk based systems or similar technologies with parallel illumination) since their field of view is fixed by the CCD chip size and the optical magnification. Ca2+ waves in muscle cells shown by a fluorescenct Calcium-indicator and recorded with a resonant scan system. Eisner University of Manchester, UK Zoom function by decreasing the scanning angle of galvanometer-mirror, pan function by adding a scanZoom 1 5 Pan offset in x and y to the (fov f iel f do With the computer controlled shifting of the y-position, the Leica TCS SP5 has a panning function also for the resonant scanner which allows for moving the position in the optical field. This is important for adjusting to a small section of the image field without cumbersome adjustment of the stage. This saves time and avoids the early damage of the sample. Drosophila melanogaster (eye section) vi e w ) zoom-image. Multicolor Imaging A further advantage of the resonant scanner is its integration into a confocal system with multiple channels and high efficient AOBS ® and SP ® detection for multi-parameter fluorescence experiments at high temporal resolution. spinning-discs) have significant limitations, which can only be compensated by simultaneous recording with multiple cameras. A very expensive and complicated approach, where multiple in fact is restricted to two. The Leica TCS SP5 en passant offers effectively 5 channels simultaneously for high speed imaging. [. . . ] Of course, one can also create nice images with a resonant scanner by line- or frame averaging. The Leica TCS SP5 merges these contradictory fields of application in a single system. This is economical on one hand, and allows to do completely different experiments with one system without having to move the sample (which would be impossible e. g. [. . . ]