User manual LEICA DM6000 CFS BROCHURE

[. . . ] Any direct manipulation of the system potentially disturbs the delicate positioning of the micropipettes within the sample. This emphasizes the importance of a remote control for the imaging setup, providing convenient access to all relevant functions. Finally, recording electrophysiological and imaging data with perfect synchrony is paramount for correct interpretation. Triggering image recording by external events and synchronizing the application of stimuli with image scanning down to the single line level helps realize sophisticated experimental setups. [. . . ] The Leica DM6000 CFS offers maximum flexibility in a multi-user environment to cater to everyone's needs. A high precision adapter is available, allowing to replace the single objective with an electronic 6x objective nosepiece. The patented change of objectives works vibration-free, with automatic power switch-off to avoid disturbing measurements. Thus, by simply pushing a button, a quick change between the magnifications can be achieved even in the near infrared without losing the area of interest, implementing automatic parfocality. Exchangeable nosepiece Features and Benefits · · · · · High precision adapter for switching between single objective and objective revolver 6 objective nosepiece positions Patented electronic nosepiece turret Automatic power switch-off after objective change Automatic electronic parfocality Top: Zebrafish eye (courtesy of: Carl Neumann, EMBL). Right (from top to bottom): Platynereis larva (courtesy of: Raju Tomer, EMBL, Heidelberg, Germany). Neurons in brain slice (courtesy of: Thomas Nevian, Institute of Physiology, Bern, Switzerland). Elisabeth Ehler, King's College, London, UK). 5 Features and Benefits of Gradient Contrast Orientation and Contrast Tissue or brain slices up to a thickness of several hundreds of microns can be optimally imaged with infrared illumination. A specially designed infrared illumination filter in combination with an IR polarizer, IR analyzer and the infrared differential interference contrast (DIC) prisms give extremely good resolution even in the thickest specimens. Used in combination or separately, fluorescence and DIC are great techniques for patch clamping. However, to avoid having any optical components in the fluorescent light path and ensure the highest photon collection efficiency for two-photon excitation fluorescence microscopy, the Dodt gradient contrast technique can also be used. This gradient contrast converts the phase information into an amplitude contrast. To study the fundamental properties of basal dendrites via patchclamp recordings, it is now possible to combine two-photon excitation fluorescence microscopy with a scanning version of this technique, called infrared-scanning gradient contrast (IR-SGC). The infrared excitation laser light and the fluorescent light are separated by a dichroic mirror, underneath a high NA condenser. The fluorescence light is detected by Non-Descanned Detectors (NDD) and the IR-scanning gradient contrast images are detected by spatially filtering the forward scattered infrared laser light with a Dodt tube and subsequent detection by a photomultiplier tube. This allows the online-overlay of a highly contrasted IR image of a brain slice with the fluorescence image of the neuron system. This detection method is patented by a Leica patent: US 6, 831, 780 B2. · · Usable both with camera (IR-video microscopy) and scanner (IRscanning gradient contrast, IR-SGC) Optical elements are outside the fluorescent light path, allowing highest possible photon collection efficiency of two-photon microscopy · · Alignment-free overlay of IR-SGC and fluorescence images Scanner patching with IR-SGC ­ no need to patch in camera mode Optical light path of Dodt gradiant contrast 6 Detection efficiency As with multi-photon excitation the fluorescence is only generated in the diffraction limited focal volume, the detectors can be placed directly behind the objective (reflected light detectors, RLD) as well as directly behind the condenser (transmitted light detectors, TLD) without losing spatial resolution. This close-coupling detection scheme results in the highest possible photon collection efficiency, as scattered fluorescent photons can also be collected over a large detection angle due to the high numerical aperture of the objective and the condenser. Two-channel detectors on both sides add a maximum of detection flexibility. Apart from its high NA, the new DM6000 CFS patented turret condenser for brightfield and interference contrast provides a number of other advantages. The condenser base with condenser head 1. 4 NA oil S1 stands for highest collection efficiency, while the patented condenser base provides a watertight seal with an outlet pipe for liquid leaking from the sample. A B Features and Benefits · C Gradient contrast with camera, magnification 0. 35x (A), 1x (B), 4x (C). Online overlay of IR-SGC and fluorescence image (D). Close-coupled two-channel NDDs both in reflected and transmitted light paths · · · Electrophysiology condenser system with outlet pipe for draining Condenser base with condenser head 0. 9 NA S1 Condenser base with condenser head 1. 4 NA oil S1 for highest collection efficiency D 7 Two worlds in one Two completely different experimental requirements can be satisfied with one single system. The Leica TCS SP5 with the Tandem Scanner provides classical morphology on large samples, where high spatial resolution is required, e. g. [. . . ] +32 2 790 98 50 Fax +61 2 9878 1055 Fax +43 1 486 80 50 30 Fax +32 2 790 98 68 Fax +1 905 762 8937 Fax +45 4454 0111 Fax +33 1 47 32 85 86 Order no. of the edition: English 1593102113 LEICA and the Leica Logo are registered trademarks of Leica IR GmbH. · Industry Division The Leica Microsystems Industry Division's focus is to support customers' pursuit of the highest quality end result. Leica Microsystems provide the best and most innovative imaging systems to see, measure, and analyze the microstructures in routine and research industrial applications, materials science, quality control, forensic science investigation, and educational applications. Richmond Hill/Ontario Tel. +1 905 762 2000 Herlev Rueil-Malmaison Wetzlar Milan Tokyo Seoul Rijswijk Tel. [. . . ]