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Equipment

 

1) Confocal SP8-SMD

Inverted Leica TCS-SP8-SMD Confocal Microscope

  • Leica Inverted DMI8 microscope stand 
  • 2 PMTs, 3 HyD (hybrid detectors) 
  • CO2/temperature controlled Tokai Hit system  for  imaging of live cells
  • Single photon lasers (attenuated using acousto-optical tuning filters (AOTFs)):
  • argon laser: excitation lines at 458, 476, 488, 496 and 514 nm (blue-cyan),
  • White light laser for excitation at any wavelength in the visible range
  • 405 nm violet (near UV) laser for near UV excitation.
  • Transmitted light detection
  • Single molecule detection system (SMD) using photon counting
  • Fluorescence Lifetime imaging (FLIM)
  • Fluorescence Correlation Spectroscopy (FCS)
  • Huygens Professional software for image processing

Main uses: advanced standard and fast scanning confocal microscopy, single molecule detection, fluorescence lifetime imaging, fluorescence correlation spectroscopy. 

 

 

2) Widefield CCD IN SITU Upright  

Widefield Leica DMRXA upright microscope

  • Fluorescence filterwheel (filters: FITC, Cy3 and DAPI, dual red/green, triple color)
  • Sensys CCD camera
    motorized stage and a piezo-focusing device
  • Digital Scientific’s Smart Capture VP QUIPS FISH software on a Macintosh computer.

Main Use: fluorescence in situ hybridization (FISH) observation of cells.





3) Confocal SP2 1P-FCS 

 Leica TCS SP2 AOBS filter-free spectral confocal microscope

  • Leica DMIRE2 inverted microscope stand, fully motorized
  • 5 photomultipliers (PMTs)
  • Differental interference contrast (DIC)
  • 2 avalanche photodiodes (APDs),
  • CO2/temperature controlled enclosure for time-lapse imaging of live cells
  • Single photon lasers (attenuated using acousto-optical tuning filters (AOTFs)):
    argon laser:excitation lines at 458, 476, 488, 496 and 514 nm(blue-cyan),
  • 3 helium-neon lasers: excitation lines at 543 (green), 594 (orange) and 633 (far red) nm
  • 405 nm violet (near UV) laser for near UV excitation.
  • Acousto-Optical Beam Splitter (AOBS): gives near instantaneous beam-switching between PMTs allowing different channels to be scanned sequentially between lines, almost mimicking simultaneous scanning without the crosstalk or bleedthrough between channels.
  • Leica Microsystems Leica Confocal Software (LCS),
  • Software modules for fluorescence (or Förster) resonance energy transfer (FRET) and fluorescence recovery after photobleaching (FRAP).
  • Separate computer and software to which fluorescence data can be exported for Fluorescence Correlation Spectroscopy (FCS) analysis (APD’s required).

Main uses: standard confocal laser-scanning microscopy over the full visible spectrum and fluorescence correlation microscopy.





4) Confocal SP2 MP-FLIM

Confocal and Multiphoton Leica TCS SP2 MP AOBS microscope system

  • Leica DMIRE2 inverted microscope stand
    Combined multi-photon/fluorescence lifetime imaging 
  • Ultrafast pulsed Spectra-Physics Mai-Tai Ti:S laser (tunable from 710 to 990 nm for IR excitation).
  • 2 non-descanned detectors (NDDs).
  • Single photon lasers (attenuated using acousto-optical tuning filters (AOTFs)):
  • argon laser:excitation lines at 458, 476, 488, 496 and 514 nm(blue-cyan),
  • 3 helium-neon lasers: excitation lines at 543 (green), 594 (orange) and 633 (far red) nm 
  • Software modules for fluorescence (or Förster) resonance energy transfer (FRET) and fluorescence recovery after photobleaching (FRAP).
  • Acousto-Optical Beam Splitter (AOBS)
    Computer attached with Becker-Hickl software for time correlated single photon counting (TCSPC) for FLIM data collection and analysis.

Main uses: standard confocal microscopy with slides and non-standard specimens, multiphoton microscopy and fluorescence lifetime imaging



5) Confocal SP5-Blue

Inverted Leica TCS-SP5 AOBS Confocal Microscope

  • Leica Inverted DMI6000 microscope stand 
  • DIC, 6 PMTs,
  • CO2/temperature controlled enclosure for time-lapse imaging of live cells
  • Single photon lasers (attenuated using acousto-optical tuning filters (AOTFs)):
  • argon laser:excitation lines at 458, 476, 488, 496 and 514 nm (blue-cyan),
  • 3 helium-neon lasers: excitation lines at 543 (green), 594 (orange) and 633 (far red) nm
  • 405 nm violet (near UV) laser for near UV excitation.
  • Acousto-Optical Beam Splitter (AOBS)

Main uses: standard and fast scanning confocal microscopy through the visible spectrum 





6) Confocal SP5 MP-STED

Leica TCS-SP5 AOBS Confocal Multiphoton STED microscope

Leica Inverted DMI6000 microscope stand

  • Set up for stimulated emission depletion (STED) microscopy for fluorescence scanning imaging below the diffraction limit of light resolution in two different color channels.
  • STED scanning uses two beams, one generated by stimulated light emission to deplete the fluorescence of certain fluorochromes for a fraction of a second. stimulated emission depletion (STED) beam at 750 nm, generated by attenuating a pulsed infrared beam from a MaiTai laser through a 100 meter fiber optic. This beam can be shaped in the form of a doughnut whose hole can be made smaller than the 200-300 nm limit of light resolution. A second beam from a either a 561 or 635 nm pulsed laser for excitation is aligned with the latter beam to pass through the hole and both beams are then be scanned across the specimen giving a much smaller effective excitation beam allowing resolution of details between 60-90 nm.
  • Resonant scanner capable of 8000 Hz scanning speed (16000 Hz with bidirectional mode)
  • single photon lasers:
    a) argon with 458, 476, 488, 496 and 514 nm excitation lines,
    b) 561 nm DPSS (yellow-green)
    c) 594 and 633 nm helium neon lasers.
  • DIC, 6 PMTs, 2 APDs, 2 NDDs 
  • CCD camera (for viewing IR)
  • MaiTai laser can also be run as a two photon laser for multiphoton scanning.
  • Software: Leica Application Suite--Advanced Fluorescence (LAS AF)

Main uses: Stimulated emission depletion pulsed scanning superresolution microscopy, confocal scanning microscopy of large and thick specimens





7) Confocal MicroALEX-FCS

A home-built single-molecule microscopy set-up for alternating laser excitation spectroscopy

  • confocal or wide-field geometry
  • Olympus IX71 microscope stand
  • 3 alternating lasers (blue, green and red) for FRET-based measurement in solution
  • Alternating Laser Excitation spectroscopy: ALEX
  • 3 APD detectors
  • fast time-correlated counting board (6 ns-resolution)
  • EMCCD camera and PC computer for recording and processing data.
  • Data can be plotted with stoichiometry vs. FRET to show different fluorochrome FRET combinations much like a plot from a flow cytometer.

Main use: alternating laser excitation spectroscopy:FRET ratioing and stoichiometry measurements at very low light levels





8) Confocal Spinning Disk

Leica DMI6000 inverted microscope with:

  • Yokogawa CSU-X1 spinning disk confocal scanner unit
  • Software controlled motorized stage and piezo-focusing device for 3-D sectioning
  • Andor EMCCD camera.
  • Solamere Technologies laser bank, computer, Yokogawa scanner and computer with running software.
  • Laser bank: 4 solid state lasers with excitation lines at 405 (dark blue) 491 (blue-green), 561 yellow-green)and 640 nm (far red).
  • Fast PC computer with over a terabyte of data storage space.
  • Running software: MicroManager opensource microscopy management software. Ultra-fast image acquisition (up to 1000 fps in full-frame) with minimal photobleaching and phototoxicity.
  • System is being tested with a Photronics CMOS super-fast camera (? frames per second) being developed at UCLA by one of our collaborators for recording of very fast events.

Main use: fast confocal scanning of live or fast moving specimens





9) Wide-Field CCD Microinjection Inverted 

Leica DMIRBE inverted microscope

  • fluorescence and DIC capabilities controlled by Ludl and Leica filterwheels,
  • Hamammatsu ORCA CCD camera
  • heated chambered stage
  • piezo-focusing device for 3-D sectioning,
  • PC computer with Slidebook software for 3-D fluorescence imaging of live cells
  • Eppendorf microinjector/ micromanipulator and second small CCD camera and for cell microinjection.

Main Use: Cell microinjection





10) In vivo Maestro 2

Cambridge Research and Instrumentation (CRi) Maestro™ 2 in vivo small animal Imaging System

  • fluorescence observation and spectral unmixing of whole mice.
  • Same LCTF technology as the NUANCE system, but applied to whole living mice in the Maestro
  • Xenon arc lamp for excitation and 7 excitation/emission sets for 500 nm to 950 nm imaging
  • Dynamic Contrast Enhancement application for image rendering.
  • PC with CRi Maestro software similar to CRi NUANCE software

Main use: spectral unmixing imaging of fluorescence signals in small animals or other macroscale specimens






11) Nikon Microscope

Nikon TE2000E inverted microscope with;

  • widefield imaging, DIC, and darkfield imaging.
  • General functions include motorized stage and autofocus.
  • dry and oil-immersion high numerical aperture specialized objectives.
  • Magnification from 20X to 100X.
  • Light sources: mercury arc lamp, and halogen lamp.
  • Cameras include Hamamatsu EMCCD for low light applications and Orca  
    Flash 2.8 for high resolution applications with frame rates of 32 and  
    45 frames per second at full resolution respectively.
  • High speed computer runs softwares that include NIS Elements by Nikon  
    and Hamamatsu HCimage.
  • System is currently being set up and tested.  

Main use: Widefield and high resolution darkfield microscopy

 

12)  Leica LMD7000 Laser Microsdissection System

Leica's latest system for laser microdissection. Laser microdissection uses a microscope to  visualize individual cells or cell clusters. Regions of interest are selected by software, excised from the surrounding tissue by a laser, and collected by gravity into specialized devices for analysis.

  • 349 nm adjustable pulse laser (max 120uJ)
  • Leica high end upright research microscope w/ motorized stage     
  •  Autofocus
  •  Color and B&W cameras
  •  Collection by gravity w/ tubes
  •  Fully automated inspection mode
  •  Fluorescence, DIC. Brightfield imaging modes
  •  Objectives 1.25X/0.04, 5X/0.12, 10X/0.3, 20X/0.4, 40X/0.6, 63X/0.7, 150X/0.9
  •       Fluorescence filters Y5 (far red), GFP for LMD, LMD–BGR triple band

Main use:  Microdissection of cells and tissue elements for downstream analysis