Light Microscopes
Acquired in 2024, funding for this instrument was provided by Canada Foundation for Innovation (Biosciences Research Infrastructure Fund) and Alberta Government infrastructure awards to Dr. Tom Hobman.
STELLARIS 8 is a true confocal point scanning system, including a White Light Laser as excitation light source, an Acousto Optical Beam Splitter (AOBS) and a highly sensitive, prism-based spectral detection design with computer controlled adjustable bandwidth for all fluorescence channels. STELLARIS 8 offers enhanced detection efficiency. The extended detection range up to 850 nm plus the expanded excitation range in the visible from 440 nm up to 790 nm allow the application and separation of an extended range of spectrally overlapping fluorophores, up to five simultaneousely. Including TauSense, a set of tools based on fluorescence lifetime information, it provides an additional contrast, improved image quality and separation of spectrally overlapping flurophores.
LIGHTNING allows for optimal extraction of image details and maximum resolution, thus expanding the imaging portfolio both in the classical range and beyond the diffraction limit.
Features a tandem scanning system to provide highest axial resolution with galvanometric scanner, switchable between resonant and non-resonant mode.
Equipped with Power HyD S detector, new standard for detection. All-round detector, high performance throughout the spectrum, two times increased photon detection efficiency in the blue-green range compared to standard multialkaline photomultiplier. Leica Si-based technology is compatible with TauSense in combination with the white light laser.
Includes Power Hybrid Detector HyD X: Specialized for Tau-STED in the visible up to far-red (depending on configuration). Leica GaAsp-based technology.
Compatible with TauSense technology in combination with the white light laser.
Inverted research microscope with touch screen based on the DMi8 series, with motorized 6x nosepiece, closed loop, cooling system and spill protection. Quick switching from coarse to fine drive. Prepared for confocal imaging with laser safety equipment.
Based around a Leica inverted DMI8 CS Premium inverted microscope base, our Stellaris 8 has the following configuration:
- Lenses: 10x/0.4 NA, 20x/0.75 NA, 40x/1.3 NA Oil, 40x/1.25 NA Glyc.(motorized), 63x 1.4 NA Oil, 63x/1.2 Water (motorized, 0.22 mm WD)
- Transmitted light modes: BF, Pol, DIC
- 6 detectors, 3 HyD S and 2 HyD X detectors, each equipped with spectral detection and tau sense detection, transmitted light PMT.
- 405 nm Diode laser
- White Light Laser (440-790nm in 1nm increments)
- Acousto-Optical Beam Splitter
- Tunable Filters
- Z Super Galvo stage
- TokaiHit stage top incubation system
- LED Fluorescence lamp
- AFC Hardware autofocus
- Resonant scanner
- LAS X Software with Dye Finder, Navigator and Lightning
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 detectors, 3 standard PMTs and 2 HyD detectors, each equipped with spectral detection
- 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
Purchased from Quorum Technologies this spinning disk confocal has the following configuration:
- Olympus IX-81 motorised microscope base
- Yokagawa CSU 10 (#1) spinning disk confocal scan-head
- Sedat Dichroic with reflections at 405/491/561/640 for standard 4-colour imaging
- Lenses: 20X/0.85 Oil, 40X/1.3 Oil, 60X/1.42 Oil
- Illumination:
- X-Cite-Mini+ LED Fluorescent Light Source for eyepiece visualization
- LMM5 from Spectral Applied Research for laser merging.
- 44mW 405nm pumped diode laser (for blue dyes, e.g. DAPI)
- 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 Orca-Fusion BT Digital sCMOS
- 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
Purchased from Quorum Technologies this spinning disk confocal has the following configuration:
- Olympus IX-81 motorised microscope base
- Yokagawa CSU-X1 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: 20X/0.85 Oil, 40X/1.3 Oil, 60X/1.42 Oil, 100X/1.4 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)
- DAPI (460/50)
- GFP (525/50)
- TRITC (593/40)
- Texas Red (620/60)
- RFP (650/100)
- Cy5 (700/75)
- "Live" filter wheel (for use with "live" samples)
- CFP (470/40)
- YFP (545/40)
- DsRed (593/40)
- mCherry (641/75)
- Cy5 (700/75)
- Empty position
- 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.
Funding for this instrument was provided by Canada Foundation for Innovation and Alberta Government infrastructure awards to Dr. Michael Hendzel.
With Lattice SIM, the sample area is illuminated with a lattice spot pattern instead of grid lines as in conventional SIM. This leads to a dramatic increase in imaging speed. In addition, the lattice pattern provides higher contrast to allow a more robust image reconstruction. Since the sampling efficiency of lattice pattern illumination is 2× higher compared to classic SIM, you need less laser dosage for sample illumination. This lattice illumination makes SIM a preferred live cell imaging technique. The strongly improved photon efficiency of lattice illumination allows you to increase the imaging speed while achieving higher contrast and lower photo dosage.
- High-powered CW lasers: 405nm, 488nm, 561nm, 642nm
- Incubation chamber for adjustable temperature, humidity, and CO2 control.
- Lenses: 63x/1.4 Oil for structured illumination, 63x/1.46 Oil for TIRF and SMLM, 63x/1.2 Water for SIM on thick / highly scattering samples, 40x/1.3 Oil for SIM Apotome mode/thick samples, 25x/0.8 LD multi-immersion lens with correction collar, 5x/0.16 Dry.
- 2 Duolinked sCMOS cameras for fast, 2-channel simultaneous acquisition (light is separated by a set of dichroics = no moving parts). Can also acquire 4 channels sequentially.
- Burst mode processing allows for SIM imaging up to 255 fps for 2D short-duration but very fast time lapse acquisition.
- Acquisition in Leap mode accelerates the volume (z) imaging speed by three times and at the same time decreases the light dosage on your sample.
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
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, DIC analyzer cube, and QUAD cube for fast DAPI/GFP/Cy3/Cy5
- Colibri LED light sources 365, 470, 555, and 625
- HXP120 metal halide source
- Motorized xy stage with holders for slides 3x1", dishes 35-60mm, and multiwell plates
- Axiocam HRm monochrome cooled CCD camera and Axiocam 105 color camera for imaging histological stains.
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, 20x/0.4 Phase LD, 20x/0.8 PA, 40x/1.3 PA Oil, 63x/1.4 PA Oil, and 100x/1.3 PN Oil.
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 (360/40 excitation, 400 Dichroic, 470/40 emission)
- for AlexaFluor488 (470/40 excitation, 495 dichroic, 525/50 emission) (Also works for other common Green dyes, e.g. FITC, GFP)
- for AlexaFluor594 (560/40 excitation, 595 dichroic, 645/75 emission) (Also works for some common red dyes, e.g. AF568, DsRed, Cy3m TRITC, Texas Red)
- for AlexaFluor647 (620/60 excitation, 660 Dichroic, 700/75 emission) (Also works for other far-red dyes, e.g. Cy5, Atto647N)
- Triple B/G/R cube for fast acquisition ( 400/20; 495 /15; 570/20 excitation, 415; 510; 590 Dichroic, 465/20; 530 /30; 640/40 emission)
Funding for this instrument was provided by Canada Foundation for Innovation and Alberta Government infrastructure awards to Dr. Meghan Riddell.
The Cell Discoverer 7 is a closed-box, inverted microscope that is ideal for live-cell imaging. The fully contained incubation system allows for stable environmental control of temperature, humidity, and CO2 over long time periods. This microscope has motorized correction collars, a sample finder, and thickness detector, so it can automatically compensate for different sample holders (multi-well plates, slides, chambers etc.), sample thicknesses, and immersion media.
Illumination modes: Fluorescence and brightfield
Lenses: 5x/0.25NA (Dry, 5.1mm working distance), 20x/0.7NA (Dry, 2.2mm working distance), 50x/1.2NA (water immersion, 0.84mm working distance)
Optovar: 0.5x, 1x, 2x
Excitation wavelengths: Colibri light sources with the following LEDs:
- 385 (DAPI / Hoechst)
- 470 (GFP / AF488)
- 567 (TdTomato / AF568)
- 625 (Cy5 / AF647)
Emission Filters:
- 425/30
- 514/30
- 592/24
- 681/45
Camera: Axiocam 712 monochrome sCMOS
Other features:
- Automatically applied/removed objective (water) immersion.
- Sample finder, providing image of sample and holder and automatic focusing.
- Motorized correction collars, enhancing imaging in plastic well-plates (5x, and 20x only), and thick samples.
- Plate adapters for holding multi-well plates, 35mm petri dishes, 60mm petri dishes, standard slides (76x26 mm), and Lab-Tek chambers (57x26mm).
- Live cell incubation (CO2, humidity, and temperature control).
- Pipette insert port for adding solutions without disturbing environmental conditions.
- Separate analysis workstation for image processing, and deconvolution.
Funding for this instrument was generously provided by the Striving for Pandemic Preparedness Alberta Research Consortium (SPP-ARC).
Historically, widefield microscopy has not been best suited for the imaging of large samples/specimen volumes. The image background, mainly originating from out-of-focus regions of the observed sample, significantly reduces the contrast, the effective dynamic range, and the maximal possible signal-to-noise ratio (SNR) of the imaging system. Instant Computational Clearing (ICC) is the core technology in THUNDER Imagers. It detects and removes the out-of-focus background for each image, making the signal of interest directly accessible. At the same time, in the in-focus area, edges, and intensity of the specimen features remain. In addition to ICC, the THUNDER microscope can perform an immediate post-processing, decision-mask- based 3D deconvolution dedicated to either thin samples (small volume computational cleaning - SVCC) or thick samples (large volume computational clearing - LVCC).
Illumination modes: Fluorescence, Brightfield, DIC, Polarized Light
Lenses: 2.5x/NA (Dry), 10x/0.3NA (Dry), 20x/0.75NA (multi-immersion), 40x/0.95NA (Dry), 63x/1.4NA (Oil immersion).
Excitation wavelengths: 8 discrete light sources with the following LEDs:
- 390 (DAPI / Hoechst)
- 440 (CFP)
- 475 (GFP / AF488)
- 510 (YFP)
- 555 (TdTomato / AF568)
- 575 (RFP / AF594)
- 635 (Cy5 / AF647)
- 747 (Cy7 / AF750)
Emission Filters:
- Quad bandpass filter cubes:
- DFT5 (435/30, 519/25, 594/32, 695/58)
- CYR7 (473/22, 539/24, 641/78, 810/80)
Camera: Leica K8 Monochrome sCMOS
Other features:
- Navigator function for fast whole specimen overview and tilescanning.
- THUNDER Live allows computationally cleared images to be viewed instantly in the live viewer to allow you to optimize ICC parameters by using immediate image feedback.
- Live cell imaging capabilities with various sample holder stage inserts for multi-well plates, 35mm petri dishes, 60mm petri dishes, standard slides (76x26 mm), as well as CO2, humidity, and temperature control.
- External emission filter wheel for “clean-up” of emission detection.
- Leica’s image analysis software (Aivia) available for image processing, analysis and segmentation workflow, to extract data from your images.
Funding for this instrument was generously provided by the University Hospital Foundation.
Creates a virtual slide image by digitizing the entire sample on the slide. The system works with both brightfield and fluorescence contrast, and can accommodate 100 standard sized slides at a time. User designed scan profiles enable unattended, automated scanning. Our system can digitise up to 5 fluorescence channels (e.g. DAPI, AF488, AF568, AF647 and AF750). Fluorescence images are captured on a state of the art Hitachi sCMOS camera (Orca Flash 4.0), while full colour images from histological colorimetric stains are captured by a progressive scan Hitachi HV-F202SCL 3X CCD camera. Fluorescence excitation is provided by an integrated, multi wavelength LED light source with excitation wavelengths of 385nm, 469nm, 555nm, 631nm and 735nm. Our system has 3 scanning lenses, 10X, 20X and 40X that give images with pixel sizes of 440nm, 220nm and 110nm, respectively. Shading correction as well as other post-acquisition processing and analysis is provided by a nearline image processing/analysis computer, optimised to view, interrogate, analyze and process the exceptionally large sized files (typical pixel dimensions: 100,000 X 100,000) that the slide scanner generates.