User manual LEICA TCS SP5 AOBS BROCHURE

[. . . ] Here, a broadband confocal microscope ­ covering all requirements by a single device ­ means a true relief. Light ­ a precious matter Sensitivity tips the scales Efficiency is the key to success. Not only in science and economy ­ where it turns to profitability, but as well in research and routine laboratories, where productivity has always been a strong requirement. Efficiency strongly depends on the suitability and the capabilities of the instrumentation. Here, first and foremost, efficiency is determined by the sensitivity of the detection device. [. . . ] In brief, the new device is not a specially coated mirror, but a switching valve for light, which is tunable to channel any laser line onto the sample and simultaneously transmit very efficiently the emitted light to the detector. It consists of an acousto optical crystal, known as tunable deflection device. For details, you may want to consult the suggested readings. Drosophila melanogaster Green: Feb211 positive neurons and their axons, Alexa 488 Red: Fibrous part of the cns (i. e. all axons), Cy3 Blue: Nuclei, DAPI Grey: Nuclei of neurons, Alexa 594 The benefits of AOBS® How does the AOBS® improve your scientific work?The transmission of the AOBS® is superior to most dichroic mirrors over the full visible spectrum. Bright and crisp images require wide emission bands as provided by the AOBS®. This is important to channel as much photons as possible from the sample to the detector ­ again improving the image quality. Low bleaching during image acquisition is important to protect the sample from fading and to protect living specimen from toxic chemicals that accumulate on photolysis of fluorochromes. The AOBS® has very steep slopes allowing collecting emission very closely to the excitation band. Any visible-range dye can be excited, as the position of the reflection-pins can be tuned individually. Multiparameter fluorescence is solved: up to eight laser lines programmable, leaving still sufficient space for emission collection ­ and the frequencies are tunable!for Ca 2+, membrane potential, pH or chloride expect fast switching in sequential scanning. The very strong suppression of the excitation can be reduced individually, if necessary for reflection imaging. ROI-scanning is improved as well: different excitation patterns are possible for different regions during a single scan. Large 3D volume recording, in sequential mode will benefit as well from fast switching devices, as speed improves dramatically the system efficiency. Fluorescence correlation spectroscopy (FCS) requires very low background and stray-light. Spectral recording (lambda scan) supply correct spectra, as the transmission of the AOBS® is "white", which means, that is does not alter the emission spectra ­ a common problem, if spectral scanning is done in a dichroic-mirror system. True confocal optical sectioning requires point-shaped illumination and emission. [. . . ] The Leica SP® overcomes the restrictions of classical filter cascades for emission splitting. It is a series of tunable elements at very high transmission, allowing selecting any wavelength band for emission collection. Tunable emission bands fit perfectly to tunable dichroics: the combination of these two technologies will not leave open any application requirement. Borlinghaus: The AOBS: Acousto Optical Beam Splitter ­ colorful brightness in confocal microscopy. [. . . ]