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O-MUG

Comprehensive Course on Fluorescence Microscopy


Raman Microscope


Built around a Leica DMI6000 automated epifluorescence microscope, the Renishaw confocal Raman microscope allows spatially resolved spectroscopy to be carried out in a confocal manner at high speed. The system can acquire Raman or PL spectra using any one of 3 solid state excitation sources (532nm, 638nm, 785nm) and a deep-depletion silicon CCD detector which has sensitivity from approximately 400-1000nm. The inverted microscope base allows for imaging of live cells or animals in typical environmental chambers which fit the Prior H117 mapping stage. All optical components (holographic notch filters, gratings, beam expansion/linefocus lenses, polarizers) are motorized and encoded, allowing for automated collection of multiple experimental conditions serially.

Model
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Renishaw inVia Confocal Raman Spectrometer
Leica DMI6000 epifluorescence microscope

Objective Lenses
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HCX PL FLUOTAR 5x/0.15
HC PLAN APO 20x/0.70
HCX PL APO 40x/0.85 CORR
HCX PL APO 63x/1.20 W CORR

Illumination Source
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Solid state lasers: 532nm (100mW), 638nm (50mW), 785mW (250mW), with matched polarizer/analyzer optics for each line
Leica EL6000 external metal halide fluorescence light source

Detection
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1024x256 deep depletion RenCam CCD for spectroscopy
600, 1200, 1800l/mm gratings on kinematic mounts
Lumenera 3-1M cooled monochrome CCD for epifluorescence documentation
DAPI, GFP, DsRed, TxRed, Cy3, YFP filter cubes

Acquisition Software
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Renishaw Wire 3.0

Advantages and Techniques
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- Fully loaded epifluorescence microscope, with ability to plan spectral mapping on fluorescence images
- Extremely high speed Raman mapping (<30ms/spectrum) possible at diffraction limited resolution
- Capable of polarization sensitive measurements, pump probe detection (532/785 or 638/785), and programmable dynamic/time lapse acquisition
- Linefocus optics available for low power density acquisition at high NA
- Arbitrary mapping pixel size for macro scanning over large areas

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