Light Microscopy

Confocal Microscopes

1. Nikon A1Rsi Confocal Microscope

The Nikon A1Rsi is a scanning confocal microscope is suitable for a broad range of applications including spectral and rapid imaging. The system is based on a motorized inverted Nikon Ti-E microscope frame equipped with the PerfectFocus stability mechanism. The system is equipped with a unique hybrid scan head, incorporating both a high-resolution galvo scanner and an ultra-high speed resonant scanner, enabling simultaneous photo-bleaching / photo-activation and image acquisition. The scan head is also equipped with two GaAsP PMTs, and two high-sensitivity PMT detectors for four-color confocal imaging as well as a transmitted light PMT for simultaneous DIC imaging. The enhanced spectral acquisition capabilities include a 32-channel detector for spectral acquisition at full image (512 x 512 pixel) resolution in 0.6 seconds.

System configuration

  • Laser lines: 405, 445, 488, 514, 561 and 640nm
  • Objectives: 10X and 20X dry and 40X, 60X and 100X high NA oil immersion
  • Motorized stage: xy-stage including a nano-positioning piezo z-insert
  • Incubation: Tokai-hit stage-top incubator capable of temperature, humidity, CO2 and O2 regulation

Applications

  • Multi-color confocal microscopy
  • DIC transmitted light microscopy
  • Three-dimensional reconstructions
  • Spectral imaging / separation
  • Co-localization analysis
  • FRAP / FRET analysis
  • Photo-activation / photo-stimulation
  • Multi-position imaging / tiling
  • Live Cell imaging (T, H, CO2 and O2)

2. Nikon Spinning Disk Confocal Microscope

The Nikon Spinning Disk (SD) confocal microscope is designed for high-magnification 4D imaging of living cells with minimal effects of photo-damage. The system is based on a motorized inverted Nikon Ti-E microscope frame equipped with the PerfectFocus stability mechanism. The system is fitted with a Yokagawa CSU-X1 variable speed Nipkow spinning disk scan head for the confocal illumination and imaging of an entire field of view (FOV) simultaneously on a camera. The scan head is equipped with an Andor TuCam two-camera adapter that enables simultaneous two-color confocal imaging at video-rate (30fps) on two separate Andor Zyla 4.2 Megapixel sCMOS cameras. The system also boasts an LED-based DMD system for ultrafast photo-stimulation and a separate galvo-based laser scanner for slower FRAP and photo-activation studies.

System configuration

  • Laser lines: 405, 488, 561 and 640nm
  • Objectives: 10X and 20X dry, 60X and 100X high NA oil immersion and 40X water immersion (corr).
  • Motorized stage: xy-stage including a nano-positioning piezo z-insert
  • Ti LAPP DMD (Deformable Mirror Device) LED source for ultrafast photo-stimulation
  • Miniscanner galvo-based laser scanning system for FRAP / photo-activation
  • Incubation: Tokai-hit stage-top incubator capable of temperature, humidity, CO2 and O2 regulation

Applications

  • Multi-color high-speed confocal microscopy
  • DIC transmitted light microscopy
  • Three-dimensional reconstructions
  • FRAP analysis
  • Photo-activation / photo-stimulation
  • Multi-position imaging / tiling
  • Live Cell imaging (T, H, CO2 and O2)

3. Olympus FV1200 Confocal Microscope

The Olympus FV1200 scanning confocal microscope is a versatile imaging platform for both fixed and living samples based on an upright BX61WI frame. The FV1200 scan head is equipped with two GaAsP PMTs and three high-sensitivity PMT detectors (2 spectral and 1 filter-based) for five-color confocal imaging, and a transmitted light PMT for simultaneous DIC imaging. The spectral PMTs are capable of acquiring spectral image data at 2nm resolution. The system is also equipped with a separate high-speed SIM laser scanner enabling simultaneous photo-bleaching / photo-activation and image acquisition

System configuration

  • Laser lines: 405, 440, 488, 515, 559 and 635nm
  • Objectives: 10X dry, 10X, 20X and 40X water immersion, 20X (corr), 100X oil immersion and 60X super chromatic aberration corrected oil immersion
  • Motorized stage: xy-stage

Applications

  • Multi-color confocal microscopy
  • DIC transmitted light microscopy
  • Three-dimensional reconstructions
  • Spectral imaging / separation
  • Co-localization analysis
  • FRAP / FRET analysis
  • Photo-activation / photo-stimulation
  • Multi-position imaging / tiling

Two-Photon Microscope

1. Zeiss LSM 880 Airyscan Confocal / Two-Photon Microscope

The Zeiss LSM 880 Airyscan confocal microscope is suitable for a broad range of applications including spectral and rapid imaging as well as deep tissue imaging. The system is based on a motorized inverted Zeiss Axio Observer Z1 microscope frame. The system is equipped with a unique scan head, incorporating a high-resolution galvo scanner along with two PMTs and a 32-element spectral detector as well as a transmitted light PMT for simultaneous DIC imaging. In addition, the Airyscan unit provides sub-diffraction limited imaging down to 140nm lateral resolution and the FAST beam shaper enables sub-diffraction limited imaging at up to 27 fps. The system is also equipped with two non-descanned detectors (NDDs) for two-photon imaging in thick tissue samples.

System configuration

  • Laser lines: 405, 458, 488, 514, 561 and 633nm and a Coherent Discovery laser with a tunable 680-1300nm line and a static 1040nm line
  • Objectives: 10X and 20X dry and 40X, 60X and 100X high NA oil immersion
  • Motorized stage: xy-stage including a nano-positioning piezo z-insert
  • Incubation: Pecon stage-top incubator capable of temperature, humidity and CO2 regulation

Applications

  • Multi-color confocal super-resolution microscopy
  • Multi-color two-photon microscopy
  • DIC transmitted light microscopy
  • Three-dimensional reconstructions
  • Spectral imaging / separation
  • Co-localization analysis
  • FRAP / FRET analysis
  • FCS, FCCS, ICS, RICS studies
  • Photo-activation / photo-stimulation
  • Multi-position imaging / tiling
  • Live Cell imaging (T, H, CO2 and O2)

Light-Sheet Microscope

1. Light-Sheet Fluorescence Microscope (OCPI-II)

OCPI-II is a home-built Light-Sheet Fluorescence Microscope (LSFM) based on an Objective-Coupled Planar Illumination (OCPI) implementation. This system excels at high-speed volumetric imaging of water-immersed fluorescent samples and achieves imaging rates that are hundreds or thousands of times faster than two-photon microscopy by simultaneously illuminating all pixels in a given focal plane, thereby eliminating the need to collect images by scanning one pixel at a time. This approach it is well suited to analyzing large ensembles of neuronal elements or for tracking highly dynamic signals such as cellular calcium. The microscope is equipped with two high speed cameras that allow synchronous dual-color imaging. An optimized piezoelectric positioner allows rapid z-scanning with a maximum of 800um of travel. Since most samples are not sufficiently clear to see 800um deep, the microscope offers a horizontal scanning mode for imaging along the surface of large samples (in addition to a conventional axial scanning mode).

System configuration

  • Laser lines: 405, 440, 488, 515 and 561nm for for imaging DAPI, CFP, GFP, YFP, and red fluorescent proteins, as well as for photoactivation and photoconversion
  • Objectives: (all water) 10X@0.3 NA, 20X@0.5 NA, 4X@0.28 NA , 10x@0.6 NA (Multi-immersion)
  • A 2-sCMOS camera system for the simultaneous imaging of two colors, including ratiometric imaging
  • 8 Configurable TTL outputs for synchronizing custom equipment / triggering stimuli
  • A multi-channel perfusion system with temperature control for maintaining physiological conditions
  • Micro-manipulators for simultaneous electrophysiology and imaging
  • An open, upright stage configuration allowing introduction of addifional components, such as inlets, for pharmacological agents, stimulating electrodes and a treadmill for in vivo mouse experiments
  • Imaging speed for a 500um cubic volume sampled with 5 um z-spacing: ~2 volumes per second (exact value depends on objective, forthcoming upgrades will increase this further)

Applications

  • High-speed volumetric imaging in living samples
  • Dual-color simultaneous imaging including with radiometric probes
  • Photoactivation and photoconversion with pixel-precise precision
  • Functional electrophysiology coupled with high-speed volumetric imaging

Super-resolution Microscopes

1. Nikon n-SIM Structured Illumination Microscope

The Nikon n-SIM microscope allows for sub-diffraction super-resolution imaging using high-frequency structured illumination in three-dimensions for resolutions beyond 100nm. The system is based on a motorized inverted Nikon Ti-E microscope frame equipped with the PerfectFocus stability mechanism. Fluorescence detection is provided by two Andor Zyla 4.2 Megapixel sCMOS cameras fitted to an Andor TuCam dual camera adapter, which facilitates simultaneous two-color and sequential four-color super-resolution imaging.

System configuration

  • Laser lines: 405, 488, 561 and 640nm
  • Objectives: 60X water immersion and 100X oil immersion high NA lenses
  • Motorized stage: xy-stage including a nano-positioning piezo z-insert
  • Incubation: Tokai-hit stage-top incubator capable of temperature, humidity and CO2 regulation

Applications

  • Multi-color super-resolution microscopy
  • Three-dimensional super-resolution reconstructions
  • Super-resolution co-localization
  • Multi-position imaging / tiling

2. Nikon n-STORM TIRF Microscope

The Nikon n-STORM TIRF Microscope allows imaging close to the surface of a coverslip facilitating imaging of endo- and exo-cytic processes as well membrane trafficking events. The system is based on a motorized inverted Nikon Ti-E microscope frame equipped with the PerfectFocus stability mechanism. Fluorescence detection is provided by either an Andor Zyla 4.2 Megapixel sCMOS camera when imaging in TIRF mode or an Andor iXon 897 EM-CCD when imaging in single-molecule or STORM (STochastic Optical Reconstruction Microscopy) super-resolution imaging mode.

System configuration

  • Laser lines: 405, 488, 561 and 640nm
  • Objectives: 10X and 20X dry and 60X and 100X oil immersion high NA lenses
  • Motorized stage: xy-stage including a nano-positioning piezo z-insert
  • Incubation: Tokai-hit stage-top incubator capable of temperature, humidity and CO2 regulation

Applications

  • Multi-color STORM super-resolution microscopy
  • 3D STORM super-resolution microscopy
  • Single-molecule imaging
  • FRET / smFRET
  • Super-resolution co-localization
  • Multi-position imaging / tiling

Fluorescence Microscopes

1. Zeiss Axio Imager Z2 Fluorescence Microscope

The Zeiss Axio Imager Z2 is a motorized upright brightfield / fluorescence microscope equipped with an ApoTome 2 optical sectioning grid imager. The system is equipped with an LED illumination source and filtersets for DAPI, GFP, RFP and far-red imaging as well with the relevant prisms and sliders for Differential Interference Contrast Microscopy (DIC). The system is equipped with two cameras; a Hammatsu Orca Flash sCMOS camera for fluorescence and a color CCD camera for histological imaging.

System configuration

  • Excite LED excitation source
  • Objectives: 5X, 10X, 20X dry and 60X and 100X oil and 40X water immersion (corr) lenses
  • Motorized stage: xy-stage

Applications

  • Multi-color fluorescence microscopy
  • Brightfield and DIC microscopy (inc. color camera for H&E stains)
  • Optical sectioning in fluorescence (ApoTome)
  • Multi-position imaging / tiling
  • Shuttle & Find correlative microscopy

2. Zeiss Stereo Lumar V.12 Fluorescence Stereoscope

The Zeiss Stereo Lumar V.12 is a motorized zoom fluorescence stereoscope equipped with LED illumination and filtersets for DAPI, GFP, RFP and far-red imaging as well with a white light source for transmitted light imaging. The system is equipped with two cameras, a Hammatsu Orca Flash sCMOS camera for fluorescence and a color CCD camera for histological imaging.

System configuration

  • Excite LED excitation source
  • Objectives: 1.5X NeoLumar (5 – 60x zoom) with a working distance of 30mm
  • Motorized stage: xy-stage

Applications

  • Multi-color fluorescence stereoscopy
  • Brightfield stereoscopy (inc. color camera for H&E stains)
  • Multi-position imaging / tiling

3. Zeiss Axioscan Z1 Automated Slide Scanning System (100-slide capacity)

The Zeiss Axioscan Z1 is a fully automated, 100-slide capacity, brightfield / fluorescence slide scanning system equipped with an LED illumination source and high-speed filter wheel fitted with filters for DAPI, GFP, RFP and far-red fluorescence imaging. The system is equipped with two cameras; a Hammatsu Orca Flash sCMOS camera for high-speed fluorescence and a color CCD camera for histological imaging of H&E, peroxidase and DAB stained slides.

System configuration

  • Excite LED excitation source
  • Objectives: 10X, 20X and 40X dry
  • High accuracy motorized stage: xy-stage
  • 100-slide capacity loader
  • Cassettes for 4 x 1x3 slides and 2 x 2x3 slides

Applications

  • Multi-color fluorescence microscopy
  • Brightfield stereoscopy (inc. color camera for H&E stains)
  • Automated Slide Scanning