Core Services and Equipment Resources

Light Microscopes

Leica TCS SP5 Laser Scanning Confocal Microscope

Acquired in 2010, thanks to a CFI grant to the CEGIIR at the University of Alberta
Based around a Leica inverted DMI 6000 B microscope base, our SP5 has the following configuration:

  • Lenses: 10X/0.3, 20X/0.5, 40X/1.25 Oil, 60X/1.2 Water, 100X/1.44 Oil
  • 5 PMTs, each equipped with AOTFs
  • 405 nm Diode laser
  • Argon ion laser (458 nm, 476 nm, 488 nm, 496 nm, 514 nm)
  • Green HeNe (543 nm)
  • Red HeNe (633 nm)
  • Acousto-Optical Beam Splitter
  • Z Galvo stage
Visit Leica for more information 

OMX

Applied Precision's OMX super resolution microscope that uses structured illumination to break the diffraction barrier.

  • Lasers: 405, 488, 561, 642
  • Solid state conventional illumination for: DAPI, CFP, Alexa488, YFP, mCherry, Alexa568, Alexa633, Alexa647, Cy5
  • Lenses:  60X/1.42 Oil for structured illumination or deconvolution, 60X/1.49 for TIRF
  • RING TIRF
  • 3X sCMOS cameras to simultaneously acquire 3 channels (light is separated by a set of dichroics, no moving parts). Can also be used to acquire 4 channels sequentially.
  • Two modes of microscopy possible - either structured illumination or fast live cell imaging. Fast live imaging means 10+ stacks per second - dependent on the thickness of your sample and the exposure time required to image your signal.
  • 37°C incubation and 5% CO2

Spinning Disk Confocal Microscope #1 and #2

Purchased from Quorum Technologies this spinning disk confocal has the following configuration:

  • Olympus IX-81 motorised microscope base
  • Yokagawa CSU 10 (#1) or CSU-X1 (#2) spinning disk confocal scan-head
    • Sedat Dichroic with reflections at 405/491/561/640 for standard 4-colour imaging
    • CFP dichroic with reflections at 440/491/561/640 for 4-colour imaging using CFP.
  • Lenses: 10X/0.3 Ph1, 20X/0.75 Dry (#1) 20X/0.85 Oil (#2), 40X/1.3 Oil, 60X/1.42 Oil
  • Illumination:
    • X-Cite 120 from Lumen Dynamics for visualization with eyepieces
    • LMM5 from Spectral Applied Research for laser merging.
    • 44mW 405nm pumped diode laser (for blue dyes, e.g. DAPI)
    • 40mW 440nm pumped diode laser (for CFP)
    • 50mW 491nm pumped diode laser (for green dyes, e.g. GFP, YFP, Alexa 488, FITC)
    • 50mW 561nm pumped diode laser (for red dyes, e.g. mCherry, Alexa 546, Cy3, TRITC)
    • 45mW 642nm pumped diode laser (for far-red dyes, e.g. Cy5, Alexa 633, Alexa 647)
  • Filter Cubes for use with eyepieces (BRIGHTLINE filter sets from Semrock):
  • Emission Filters for use during Confocal Imaging: The system is fitted with two interchangeable filter wheels, one for imaging standard 4 colour (Blue, Green, Red, Far-Red) “fixed” samples, and one optimised for imaging fluorophores commonly found in “live” cell experiments. Note that with these emission filters, there are no excitation filters necessary, as the lasers provide single wavelength excitation. Additionally, there is a choice of three dichroic mirrors housed in the confocal scan unit. Most commonly used, the Sedat dichroic is compatible with 4 colour fixed imaging. The second choice is a dichroic that maximises the emission from GFP (EGFP), but at the expense of separating its emission from other fluorophores. Last is a dichroic that is designed for separating CFP from YFP.
  • “Fixed” filter wheel (for use with “fixed” samples)
    • Polariser (for DIC imaging)
    • DAPI (460/50)
    • GFP (515/30)
    • Cy3 (595/50)
    • Texas Red (620/60)
    • Cy5 (690/50)
  • “Live” filter wheel (for use with “live” samples)
    • 436/24
    • CFP (470/24)
    • GFP (520/40)
    • YFP (540/30)
    • RFP (595/50)
    • Cy5 (700/75)
  • Imaging using a Hamamatsu EMCCD (C9100-13).
  • Acquisition using Perkin Elmer’s Volocity
  • Chamlide TC-A Live Cell Chamber (37°C incubator + 5% CO2 atmosphere). Can accommodate either 18mm or 25mm diameter coverslips.
  • ASI MS-2000 motorised XY stage with a piezo Z insert that has 100µm travel.

Deltavision Elite

Automated, integrated widefield, fluorescence microscope from GE Healthcare. Uses Deconvolution to increase contrast and resolution in the image.

  • Olympus IX-71 microscope base:
    • Sedat Dichroic with reflections at 405/491/561/640 for standard 4-colour imaging
    • CFP dichroic with reflections at 440/491/561/640 for 4-colour imaging using CFP.
  • Lenses: 10X/0.3,  40X/1.3 Oil, 60X/1.42 Oil
  • Illumination:
    • SSI Solid State Illumination with excitation wavelengths ~405nm, 440nm, 480nm, 561nm and 640nm
    • Analyser for DIC visualisation
  • Emission Filters:
    • “Fixed” filter wheel (for use with “fixed” samples)
      • Polariser (for DIC imaging)
      • DAPI (460/50
      • FITC (515/30
      • TRITC (595/50
      • Cy5 (620/60)
    • “Live” filter wheel (for use with “live” samples)
      • 436/24
      • CFP (470/24)
      • GFP (520/40)
      • YFP (540/30)
      • RFP (595/50)
      • Cy5 (700/75)
  • Imaging using an Edge sCMOS, up to 2048X2048 (220µm X 220µm field of view)
  • Environmental control (37°C incubator + 5% CO2 atmosphere). Can accommodate multiple sample holders.
  • Flexure stage.

Zeiss Colibri Fluorescence Microscope

Zeiss AxioObserver.Z1

  • z motor focus with 25nm step
  • Brightfield, phase contrast, DIC transmitted techniques in motorized condenser
  • 6x motorized fluorescence filter turret with DAPI/BFP, AF488/GFP, Rhod/Cy3, CY5, Cy7, DIC analyzer cube
  • Colibri LED light sources 365 470 590 with triple RGB filter cube and Texas Red, GFP, DAPI and white LED for individual filter sets
  • HXP120 metal halide source
  • Motorized xy stage with holders for slides 3x1", dishes 35-60mm, multiwell plates and 90mm dishes
  • Axiocam HRm monochrome cooled CCD camera

AxioVision imaging has MultiChannel overlay/merging, z-stack, timelapse, mark and find, Mosaix, deconvolution, 3d rendering, Apotome optical sectioning. 

Objectives: 
10x/0.3 Phase PN, 10x/0.45 PA, 20x/0.4 Phase LD, 20x/0.8 PA, 40x/0.9 dry, 40x/1.3 PA, 40x/0.6 Phase LD, 63x/1.4 PA.


Leica Widefield DMI6000 Inverted

Based around a Leica inverted DMI 6000 microscope base, this microscope has the following features:

  • Lenses:
    • 100X/1.49 Oil
    • 40X/1.3 Oil
    • 20X Dry
    • 10X Dry
    • 4X Dry
  • Fluorescence Filtersets:
    • for DAPI (365/12 excitation, 395 Dichroic, 397 nm Longpass emission)
    • for AlexaFluor488 (470/40 excitation, 510 dichroic, 540/50 emission) (Also works for other common Green dyes, e.g. FITC, GFP)
    • for Rhodamine (546/12 excitation, 560 dichroic, 608/65 emission) (Also works for other common Red dyes, e.g. Cy3, TRITC, DsRed)