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Microscopy Assets

Microscopy Assets

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Microscopes and related equipment:

  1. Two photon/confocal microscope – custom built
    1. Nikon TE2000-U stand.
    2. Halogen condenser illumination.
    3. Mercury arc epifluorescence.
    4. Variety of fluorescence cubes.
    5. Objectives:
      1. Nikon 60x/1.27NA Plan Apo IR water immersion with DIC.
      2. 40x/0.6NA Plan Fluor with phase contrast.
  • 20x/0.75NA Plan Fluor ELWD with DIC.
  1. Physik Instrumente fast piezo focuser on 60x objective.
  2. Argon ion laser (Spectra-Physics).
  3. Titanium:Sapphire laser: Mai Tai HP (Spectra-Physics).
  4. Fast Pockels cell (Conoptics) on Mai Tai beam path.
  5. Two Avalanche photodiode detectors (Perkin Elmer) – single photon counting modules.
  6. Two Hybrid detectors (Hamamatsu/Becker and Hickl).
  7. Becker and Hickl fluorescence lifetime module.
  8. x-y galvanometer mirrors (Cambridge Technology).
  9. Custom LabVIEW-based instrument control and acquisition software and hardware (National Instruments).
  10. Vibration dampened optical bench with optics and optomechanical components (various vendors).
  11. Custom applications:
    1. Quantitative subcellular protein distribution analysis
    2. 2 photon FRAPa and FRAPb
  • 2 photon FLIM-based FRET
  1. 2 photon FLIM-based polarization anisotropy microscopy
  2. Fluorescence correlation spectroscopy
  1. sptPALM and STORM microscope (super resolution).
    1. Nikon Ti2 stand.
    2. Objectives:
      1. SR HP Apo TIRF 100x/1.49 Oil.
      2. 20x/0.75NA Plan Apo.
    3. LED condenser illumination.
    4. LED EXFO epifluorescence.
    5. 3D N-STORM module with high power 4 laser system.
    6. ILAS2 ring TIRF illumination.
    7. Ixon Ultra 892 EMCCD camera.
    8. Prime BSI sCMOS camera.
    9. Live cell incubation chamber – okolab.
    10. NIS-Elements Advanced Research
      1. AI module.
      2. 3D super resolution analysis.
  • AI-mediated automated acquisition.
  1. Custom applications:
    1. sptPALM for high density mapping of single molecule dynamics in live cells.
    2. 3D sptPALM under development.
  • Qdot-based single molecule tracking in live cells.

Microscopes and related equipment:

  • Leica TIRF/dSTORM microscope (super resolution)
    • LED widefield illumination
    • Solid state 120-150 mW 405, 488, 561, and 651 lasers
    • Standard fluorescent cubes and filters (for wavelengths above)
    • Objectives:
      • 20x air
      • 40x oil pl apo 1.25 NA DIC
      • 63x pl apo 1.47 NA TIRF/DIC, oil (temperature correction)
      • 100x pl apo 1.47 NA TIRF/DIC, oil (temperature correction)
      • 163x pl apo 1.45 NA TIRF/DIC, oil (temperature correction)
    • DIC and polarization setup
    • Motorized stage
    • LASX software with STORM module
    • Andor iXon EMCCD camera
    • Antivibration table
    • Autofocus
    • Advanced peristaltic pump
    • Custom microfluidics devices
    • OKO labs heater (stage insert and objective heater) – no gas

Applications:

  • Standard epifluorescence
  • TIRF/HiLo (VAEM) for live or fixed cells
  • TIRF “biochemistry on a coverglass” reconstitution experiments
  • SNAP- Halo-tag based single molecule tracking in live cells and in vitro reconstitution assays
  • Nikon SoRa spinning disk confocal and deconvolution system (super resolution)
    1. SOLA LED widefield illumination
    2. Solid state Solid state 100 mW 405, 488, 561, and 651 lasers
    3. Photometrics Prime sCMOS camera
    4. Objectives
      1. 20x pl apo 0.75 NA
      2. 60x pl apo 1.4 NA
    5. Piezo stage
    6. Motorized turret and stage
    7. NIS-elements software
    8. SoRa deconvolution software upgrade
    9. Antivibration table
    10. Yokogawa CSU-W1

Applications:

  1. Super resolution spinning disk confocal (40-50 nM resolution)
  2. Single molecule tracking in cells

Extremely fast 4D (Z stack + time) movies

The lab houses a TIRF microscope that belongs to the CDB department. 

This is a Nikon TE2000 microscope equipped for live cell imaging (with a temperature-controlled environmental chamber). 

The following imaging modalities and objectives are available: IRM, DIC (60X and 20X), phase contrast (10X), epifluorescence (20X, 60X, 100X), and 4-color TIRF (60X or 100X). The following TIRF lasers are available: 405 nm, 488 nm, 561 nm, and 640 nm.  

The microscope is equipped with the motorized Z-position control, Perfect Focus, and with a Prime 95B cameras and is controlled by a dual monitor computer using Nikon Elements software. The microscope is equipped with filter sets for multicolor TIRF, DAPI, TRITC, GFP, and CY5. 

Instrumentation in Core

  • Zeiss LSM510 scanning confocal/two-photon microscope system
  • Zeiss LSM 780 confocal
  • Deep Brain 2p Imaging Microscope
  • Superresolution Microscope
  • Environmental chamber providing temperature and CO2 control
  • Coherent Chameleon tunable phased NIR laser for two-photon excitation
  • Newport vibration isolation optical table
  • A separate graphical workstation with Zeiss and ImageJ software for image analysis

    Website
  • Select a confocal geometry optimal for your assay and throughput needs
  • Achieve excellent image quality without sacrificing throughput via our unique optical path technology
  • Acquire statistically relevant data quickly with an advanced scientific CMOS detector, enabling >3 log dynamic range
  • Large field of view enables whole-well imaging
  • Expand your research capabilities with transmitted light, liquid handling, and environmental options

Confocal technology at the speed of widefield imaging:

  • Capture an entire well of a 384‑well plate with a single image at 4X magnification
  • Capture four wells of a 1536- well plate in a single image at 4X magnification
  • Throughput of >160K wells/day confocal, >200K wells/day widefield 2 ImageXpress

Website

  • JEM-2100 (JEOL) 80-200 kV cryo configuration
  • LaB6 electron source
  • Gatan 626 cryo stage
  • TVIPS F415-MP 4096x4096 pixel slow scan CCD camera, 15 µm pixel size, 6.1 x 6.1 cm imaging area
  • TVIPS F114-NX 1024x1024 pixel fast scan CCD camera, 14 µm pixel size, up to 26 frames/sec for life imaging
  • EMMENU software for manual and automatic image collection

Website

Leica STED/Light Sheet advanced imaging system with five channel spectral confocal: (IHP/NRB-4624B)

  1. STED (stimulated emission depletion) 3X nanoscopy
    This is a super-resolution technique that overcomes diffraction limited resolution. With multiple STED laser lines of at 592 nm, 660 nm and the pulsed laser at 775 nm, STED can reach a resolution below 30 nm.
    Key Features:
    • UV Laser 405 nm, White light laser WLL2 470-670 nm.
    • STED Laser: 592 nm, 660 nm and 775 nm.
    • STED objectives: 93x glycerol and 100x oil.
    • Five spectral detector channels: two PMTs and three HyDs.
    • Tandem scanner (8kHz).
    • XY Scanning Stage.
    • HyVolution2 for STED.
  2. Digital light sheet (DLS) Microscopy
    DLS microscopy is a fluorescence technique that uses a plane of light for 3-D imaging that is useful for large samples such as embryos, sensitive samples, or fast biological processes in vivo. It provides good optical sectioning capabilities and high speed with low levels of photobleaching.
    Key Features:
    • 4.2MP CMOS camera.
    • Detection objectives: 5x, 10x and 25x water.
    • TwinFlect mirror: 2.5 mm, 5mm and 7.8 mm.
  3. Confocal Microscopy
    Key Features:
    • UV Laser 405 nm, White light laser WLL2 470-670 nm.
    • Detection objectives: 10x dry, 20x dry, 40x oil, 63X oil.
    • Five spectral detector channels: two PMTs and three HyDs.
    • Tandem scanner (8kHz).
    • XY Scanning Stage.
    • HyVolution2 SuperResolution package

Leica SP8 confocal microscopy: (WH-108):

Leica's latest confocal system with three channel spectral detectors and HyVolution software that combines super-sensitive HyD detection with SVI Huygens deconvolution, allowing lateral resolution down to 140 nm compared with 200-250 nm for standard confocal microscopy.
Key Features:

  • UV Laser 405 nm, VIS Laser 488 nm, 552 nm and 638 nm.
  • Detection objectives: 10x dry, 20x dry and 63x Oil.
  • Three spectral detector channels: two PMTs and one HyD.
  • XY Scanning Stage.
  • HyVolution2 SuperResolution package.

Website



  • Select a confocal geometry optimal for your assay and throughput needs
  • Achieve excellent image quality without sacrificing throughput via our unique optical path technology
  • Acquire statistically relevant data quickly with an advanced scientific CMOS detector, enabling >3 log dynamic range
  • Large field of view enables whole-well imaging
  • Expand your research capabilities with transmitted light, liquid handling, and environmental options

Confocal technology at the speed of widefield imaging:

  • Capture an entire well of a 384‑well plate with a single image at 4X magnification
  • Capture four wells of a 1536- well plate in a single image at 4X magnification
  • Throughput of >160K wells/day confocal, >200K wells/day widefield 2 ImageXpress

Website

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