DM6000 CFS.qxd:DM6000 CFS - Duke University Light Microscopy ...

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Two worlds in oneTwo completely different experimental requirements can be satisfiedwith one single system. The Leica TCS SP5 with the TandemScanner provides classical morphology on large samples, wherehigh spatial resolution is required, e.g. research on structures ofcytoskeleton, organelles or tissues, as well as physiology and biophysicsimaging, where temporal resolution becomes very important.Double labeled neurons of brain slices.Courtesy of Thomas Nevian, Institute ofPhysiology, Bern, SwitzerlandCalcium is an important second messenger to trigger many signallingcascades in neurons. Functional imaging of calcium influxis possible with calcium ion sensitive indicator dyes, but calciumtransients in neurons are very fast. Therefore, they are typicallyrecorded in line scan mode, also known as xt-scan mode. Thelaser beam is continuously scanned back and forth along thesame line and fluorescence over time is recorded. The resultingimage consists of one spatial and one time axis. The resonantscanning system of the Leica TCS SP5 oscillates at 8000 Hz,enabling a line rate of 16,000 lines per second. Fast dynamics ofinitial calcium inflow can now be investigated.Even in the xy-scan mode, the scan rate can be as high as 180frames per second in the frame size 512 x 64. This frame rateenables imaging of extended regions of the dendritic tree andmultiple spines at the same time.Confocal interface8
New XYTZ-Scan Mode for 3D visualization of calcium transientsThe dendritic tree has a complex three dimensional structure.Repeatedly acquiring complete 3D stacks gives a temporal resolutionmuch too low for imaging the fast calcium transients in dendrites.To circumvent this problem, optical sectioning, the fastscanning of the resonant system and the triggering capability arecombined for the new XYTZ-Scan Mode.Individual time series are taken at different focal depths and combinedinto a 4D image stack. A stimulus is always delivered beforethe same frame of each time series, synchronized by a trigger outevent. After a complete focal series, the image data is projectedinto a 3D data stack over time. For each structure in the sampled3D volume the fluorescence transients can be analyzed. The timecourse of fluorescence in all parts of the dendritic tree can beseen clearly.XYTZ-Scan Module:Time series at different focalpositions, stimulus (t 0 ) is always delivered beforethe same frame of each time seriesScanner patching and optimized workflowIn combination with the IR-SGC, which makes electrophysiologyneedles visible in the scanning mode, the extreme speed of theresonant scanner allows electrode patching while imaging atvideo rates, without ever switching to the camera mode. Togetherwith the one-for-all electrophysiology 20x 1.0 objective, scannerpatching minimizes the number of steps between experimentsetup and data collection, optimizing your workflow.So even though you might never need it, the workflow orienteduser interface of the Leica TCS SP5 with Leica <strong>DM6000</strong> <strong>CFS</strong>includes the camera as well. Whether you are doing CCD-cameraor confocal / multiphoton imaging, the other option is always justone fast click away. Integrating both operation modes in the samesoftware allows you to concentrate on what’s in your images, noton where they came from.Features and BenefitsCamera interface• Software-integration of CCDcameraand confocal scanner• Single click switching betweencamera and confocal• Workflow oriented user interface9
Page 1 and 2: Leica DM6000 CFSConfocal Fixed Stag
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Page 11 and 12: System ComponentsLeica Confocal Fix

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