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

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Features and Benefitsof Gradient Contrast• Usable both with camera (IR-videomicroscopy) and scanner (IRscanninggradient contrast, IR-SGC)• Optical elements are outside the fluorescentlight path, allowing highestpossible photon collection efficiencyof two-photon microscopy• Alignment-free overlay of IR-SGCand fluorescence images• Scanner patching with IR-SGC –no need to patch in camera modeOrientation and ContrastTissue or brain slices up to a thickness of several hundreds ofmicrons can be optimally imaged with infrared illumination. A speciallydesigned infrared illumination filter in combination with anIR polarizer, IR analyzer and the infrared differential interferencecontrast (DIC) prisms give extremely good resolution even in thethickest specimens.Used in combination or separately, fluorescence and DIC aregreat techniques for patch clamping. However, to avoid havingany optical components in the fluorescent light path and ensurethe highest photon collection efficiency for two-photon excitationfluorescence microscopy, the Dodt gradient contrast techniquecan also be used. This gradient contrast converts the phase informationinto an amplitude contrast. Images of neurons look similarto images obtained with DIC.To study the fundamental properties of basal dendrites via patchclamprecordings, it is now possible to combine two-photon excitationfluorescence microscopy with a scanning version of thistechnique, called infrared-scanning gradient contrast (IR-SGC).The infrared excitation laser light and the fluorescent light areseparated 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 detectedby spatially filtering the forward scattered infrared laser light witha Dodt tube and subsequent detection by a photomultiplier tube.This allows the online-overlay of a highly contrasted IR image of abrain slice with the fluorescence image of the neuron system. Thisdetection method is patented by a Leica patent: US 6,831,780 B2.Optical light path of Dodt gradiant contrast6
Detection efficiencyAs with multi-photon excitation the fluorescence is only generatedin the diffraction limited focal volume, the detectors can beplaced directly behind the objective (reflected light detectors,RLD) as well as directly behind the condenser (transmitted lightdetectors, TLD) without losing spatial resolution. This close-couplingdetection scheme results in the highest possible photon collectionefficiency, as scattered fluorescent photons can also becollected over a large detection angle due to the high numericalaperture of the objective and the condenser. Two-channel detectorson both sides add a maximum of detection flexibility.Apart from its high NA, the new <strong>DM6000</strong> <strong>CFS</strong> patented turret condenserfor brightfield and interference contrast provides a numberof other advantages. The system allows the exchangebetween dry and oil condensers. The condenser base with condenserhead 1.4 NA oil S1 stands for highest collection efficiency,while the patented condenser base provides a watertight sealwith an outlet pipe for liquid leaking from the sample.ABFeatures and BenefitsDCGradient contrast with camera,magnification 0.35x (A), 1x (B), 4x (C).Online overlay of IR-SGC andfluorescence image (D).• Close-coupled two-channel NDDsboth in reflected and transmittedlight paths• Electrophysiology condenser systemwith outlet pipe for draining• Condenser base with condenserhead 0.9 NA S1• Condenser base with condenserhead 1.4 NA oil S1 for highestcollection efficiency7
Page 1 and 2: Leica DM6000 CFSConfocal Fixed Stag
Page 3 and 4: Variable Fieldat High ResolutionWit
Page 5: Large range of applicationsEven tho
Page 9 and 10: New XYTZ-Scan Mode for 3D visualiza
Page 11 and 12: System ComponentsLeica Confocal Fix

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

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