Instrumentation
| Dual Iterative Lattice Structured Illumination Microscopy (SIM2) with the Zeiss Elyra 7 |  |
|---|
Location
• Center for Translational Research and Education (CTRE), room 175
Capabilities
• The Elyra 7 Lattice SIM2 system is versatile, allows for superior temporal and spatial resolution, is more efficient, and has a larger field of view than other systems.
• The Elyra 7 Lattice SIM2 system allows users to achieve resolutions of 60 nm (X, Y) and 200 nm (Z) in a variety of samples (fixed, living cells, thick specimens) using standard staining techniques and 20-30 nm (X, Y) with single molecule localization microscopy (SMLM) techniques such as photoactivated localization microscopy (PALM), direct stochastic optical reconstruction microscopy (dSTORM), and points accumulation for imaging nanoscale topography (PAINT).
• Fixed and living specimens can be imaged gently at speeds of up to 255 frames per second.
• Wide-field system with a field of view of 80 × 80 µm with a 63× lens.
• The Elyra 7 Lattice SIM2 system incorporates several microscopy techniques including Lattice SIM2, SIM2 Apotome, widefield differential interference contrast (DIC), SMLM, and total internal reflection fluorescence (TIRF) microscopy.
• The Elyra 7 can be combined with other imaging modalities, such as transmission electron microscopy, in a correlative workflow.
• Simultaneous two-color imaging of the entire field of view.
• The use of SIM2 Apotome acquisition mode allows for fast optical sectioning with contrast and lateral and axial resolution far superior than that achievable with conventional widefield systems.
• The use of Elyra 7 Leap mode increases the speed at which a volume is imaged by three times while significantly decreasing the light dosage on the sample.
Instrument Features
• 4 diode lasers: 405, 458, 561, 642 nm
• Objectives: 10× dry, 40× oil immersion, 63× oil immersion
• Heated stage insert and controller
• Incubation chamber, CO2 cover, temperature controller, heating unit, and humidity controller
• High-speed, high-precision XYZ scanning stage (travel range: 130 × 100 mm)
• Dual high-speed scientific complementary metal-oxide-superconductor (sCMOS) cameras with integrated liquid cooling and a DuoLink adapter
• AxioObserver 7 inverted microscope with Definite Focus 3 correction unit
• Premium microscopy workstation equipped with Zen software suite with time-lapse, z-stack, tiles, multi-position imaging, 3D rendering, co-localization, and automated image analysis tools
Acknowledgment
• Users of the Elyra 7 Lattice SIM2 system are to acknowledge the support of the National Institutes of Health Shared Instrument Grant by including the following statement in presentations and peer-reviewed papers: A Shared Instrument Grant from the National Institutes of Health (grant number 1S10OD034431-01) supported this work.
| Transmission Electron Microscopy with the Philips CM 120 |  |
|---|
Location
• Center for Translational Research and Education (CTRE), room 173
Capable of collecting high quality, high resolution, 2-dimensional images of biological and non-biological specimens maintained at room temperature.
Instrument Features
• Tungsten electron gun (i.e. illumination source)
• Can be operated at 20, 40, 60, 80, 100, or 120 kV
• Equipped with twin objective lenses, a CompuStage specimen stage, and a single tilt sample holder
• Equipped with a BioSprint 16-megapixel digital camera
• Magnification range: 31× to 660,000×
| Multiphoton/Confocal Microscopy with the Leica TCS SP5 |  |
|---|
Location
• Center for Translational Research and Education (CTRE), room 174
Capabilities
• Capable of collecting low and high magnification high quality, high resolution 2, 3, 4, and 5-dimensional (X, Y, Z, time, and spectrum) images
• Short-term live cell imaging experiments
• Semi-automated collection of 2- and 3-dimensional images from multiple X, Y, and Z locations over time
• Fluorescence recovery after photobleaching (FRAP), fluorescence resonance energy transfer (FRET), photo-activation (PA), photo-stimulation (PS), and photo-uncaging
• Tile stitching and image analysis
• Deep tissue (i.e., several hundred microns) imaging
Instrument Features
• Argon, HeNe, and titanium-sapphire (TiSa) pulsed (1-1.5 picoseconds duration) lasers
• Laser lines: 458, 476, 488, 496, 514, 543, 594, 633 nm
• Objectives: 10× dry, 20× dry, 40× oil immersion, 63× water immersion, 63× oil immersion
• Filter-free spectral detector system with 5 individually tunable channels
• Each detector is freely tunable over the entire visible spectrum
• The spectral range of the detector optics covers 350-900 nm with 5 nm spectral resolution and sub-micron spatial resolution
• The scanner can be run in conventional or resonant modes
o The conventional scanner can be scanned uni- or bidirectional with speeds ranging from 10-2800 Hz (2-3 fps at 512 × 512 pixels).
o The resonant scanner can be scanned uni- or bidirectional at 8000 Hz or 1600 Hz, respectively, with a maximum frame rate of 250 fps.
• Heated stage insert and controller
• Inverted microscope stand
• Digital zoom, 360° image rotation, and imaging multiple X, Y, and Z positions
• Equipped with a Z-galvanometer that allows for XZ scanning and fast XYZ scanning
• Scanning stage allows for stitching together images from large fields of view
• Time-lapse
• Leica Application Suite Advanced Fluorescence (LAS AF) software
• High confocal speed: 28 frames per second (fps) at 512 × 512 pixels
| Image Analysis Workstation |  |
|---|
Location
• Center for Translational Research and Education (CTRE), room 175
Capabilities
• Interactive processing, visualization, and analysis of 3D and 4D microscopic images via Imaris software.
• State-of-the-art volume and surface rendering, object detection, tracking, and cell division tracking tools via Imaris software.
• Automated processing of image series in a batch.
• Automatic or semi-automatic segmentation, visualization, and quantification of filamentous structures via Imaris Filament Tracer module.
Instrument Features
• Dell Precision 7820 Tower base
• Windows 10 Pro for Workstation
• NVIDIA GeForce GTX 1080 graphic card
• Dual Intel Xeon gold 5122 3.6 GHz processors
• 128 GB (8×16GB) RAM
• Imaris 9.5 Basic Tracking with XT (XBT)
• Imaris Batch 9.5
• Imaris Filament Tracer 9.6
• Image J
| Leica EM UC7 ultramicrotome |  |
|---|
Location
• Center for Translational Research and Education (CTRE), room 176
Capabilities
• Prepares consistent, high-quality semi-thin and ultrathin sections for transmission electron microscopy and light microscopy.
Instrument Features
• Thickness feed range from 1 nm to 15 µm.
• Cutting speed range from 0.05 to 100 mm/s.
• Eucentric movement allows for optimal knife approach to the sample.
• Outstanding LED illumination (top, back, and transmitted light; independent control of the illumination modes) yields outstanding visibility of the knife edge and block face.
• Fully motorized knife stage and auto-trim function.
• Vibration-decoupled gravity-stroke cutting arm for chatter-free sectioning.
• Touchscreen control unit.
| Dual Iterative Lattice Structured Illumination Microscopy (SIM2) with the Zeiss Elyra 7 | |
|---|
Location
• Center for Translational Research and Education (CTRE), room 175
Capabilities
• The Elyra 7 Lattice SIM2 system is versatile, allows for superior temporal and spatial resolution, is more efficient, and has a larger field of view than other systems.
• The Elyra 7 Lattice SIM2 system allows users to achieve resolutions of 60 nm (X, Y) and 200 nm (Z) in a variety of samples (fixed, living cells, thick specimens) using standard staining techniques and 20-30 nm (X, Y) with single molecule localization microscopy (SMLM) techniques such as photoactivated localization microscopy (PALM), direct stochastic optical reconstruction microscopy (dSTORM), and points accumulation for imaging nanoscale topography (PAINT).
• Fixed and living specimens can be imaged gently at speeds of up to 255 frames per second.
• Wide-field system with a field of view of 80 × 80 µm with a 63× lens.
• The Elyra 7 Lattice SIM2 system incorporates several microscopy techniques including Lattice SIM2, SIM2 Apotome, widefield differential interference contrast (DIC), SMLM, and total internal reflection fluorescence (TIRF) microscopy.
• The Elyra 7 can be combined with other imaging modalities, such as transmission electron microscopy, in a correlative workflow.
• Simultaneous two-color imaging of the entire field of view.
• The use of SIM2 Apotome acquisition mode allows for fast optical sectioning with contrast and lateral and axial resolution far superior than that achievable with conventional widefield systems.
• The use of Elyra 7 Leap mode increases the speed at which a volume is imaged by three times while significantly decreasing the light dosage on the sample.
Instrument Features
• 4 diode lasers: 405, 458, 561, 642 nm
• Objectives: 10× dry, 40× oil immersion, 63× oil immersion
• Heated stage insert and controller
• Incubation chamber, CO2 cover, temperature controller, heating unit, and humidity controller
• High-speed, high-precision XYZ scanning stage (travel range: 130 × 100 mm)
• Dual high-speed scientific complementary metal-oxide-superconductor (sCMOS) cameras with integrated liquid cooling and a DuoLink adapter
• AxioObserver 7 inverted microscope with Definite Focus 3 correction unit
• Premium microscopy workstation equipped with Zen software suite with time-lapse, z-stack, tiles, multi-position imaging, 3D rendering, co-localization, and automated image analysis tools
Acknowledgment
• Users of the Elyra 7 Lattice SIM2 system are to acknowledge the support of the National Institutes of Health Shared Instrument Grant by including the following statement in presentations and peer-reviewed papers: A Shared Instrument Grant from the National Institutes of Health (grant number 1S10OD034431-01) supported this work.
| Transmission Electron Microscopy with the Philips CM 120 | |
|---|
Location
• Center for Translational Research and Education (CTRE), room 173
Capable of collecting high quality, high resolution, 2-dimensional images of biological and non-biological specimens maintained at room temperature.
Instrument Features
• Tungsten electron gun (i.e. illumination source)
• Can be operated at 20, 40, 60, 80, 100, or 120 kV
• Equipped with twin objective lenses, a CompuStage specimen stage, and a single tilt sample holder
• Equipped with a BioSprint 16-megapixel digital camera
• Magnification range: 31× to 660,000×
| Multiphoton/Confocal Microscopy with the Leica TCS SP5 | |
|---|
Location
• Center for Translational Research and Education (CTRE), room 174
Capabilities
• Capable of collecting low and high magnification high quality, high resolution 2, 3, 4, and 5-dimensional (X, Y, Z, time, and spectrum) images
• Short-term live cell imaging experiments
• Semi-automated collection of 2- and 3-dimensional images from multiple X, Y, and Z locations over time
• Fluorescence recovery after photobleaching (FRAP), fluorescence resonance energy transfer (FRET), photo-activation (PA), photo-stimulation (PS), and photo-uncaging
• Tile stitching and image analysis
• Deep tissue (i.e., several hundred microns) imaging
Instrument Features
• Argon, HeNe, and titanium-sapphire (TiSa) pulsed (1-1.5 picoseconds duration) lasers
• Laser lines: 458, 476, 488, 496, 514, 543, 594, 633 nm
• Objectives: 10× dry, 20× dry, 40× oil immersion, 63× water immersion, 63× oil immersion
• Filter-free spectral detector system with 5 individually tunable channels
• Each detector is freely tunable over the entire visible spectrum
• The spectral range of the detector optics covers 350-900 nm with 5 nm spectral resolution and sub-micron spatial resolution
• The scanner can be run in conventional or resonant modes
o The conventional scanner can be scanned uni- or bidirectional with speeds ranging from 10-2800 Hz (2-3 fps at 512 × 512 pixels).
o The resonant scanner can be scanned uni- or bidirectional at 8000 Hz or 1600 Hz, respectively, with a maximum frame rate of 250 fps.
• Heated stage insert and controller
• Inverted microscope stand
• Digital zoom, 360° image rotation, and imaging multiple X, Y, and Z positions
• Equipped with a Z-galvanometer that allows for XZ scanning and fast XYZ scanning
• Scanning stage allows for stitching together images from large fields of view
• Time-lapse
• Leica Application Suite Advanced Fluorescence (LAS AF) software
• High confocal speed: 28 frames per second (fps) at 512 × 512 pixels
| Image Analysis Workstation | |
|---|
Location
• Center for Translational Research and Education (CTRE), room 175
Capabilities
• Interactive processing, visualization, and analysis of 3D and 4D microscopic images via Imaris software.
• State-of-the-art volume and surface rendering, object detection, tracking, and cell division tracking tools via Imaris software.
• Automated processing of image series in a batch.
• Automatic or semi-automatic segmentation, visualization, and quantification of filamentous structures via Imaris Filament Tracer module.
Instrument Features
• Dell Precision 7820 Tower base
• Windows 10 Pro for Workstation
• NVIDIA GeForce GTX 1080 graphic card
• Dual Intel Xeon gold 5122 3.6 GHz processors
• 128 GB (8×16GB) RAM
• Imaris 9.5 Basic Tracking with XT (XBT)
• Imaris Batch 9.5
• Imaris Filament Tracer 9.6
• Image J
| Leica EM UC7 ultramicrotome | |
|---|
Location
• Center for Translational Research and Education (CTRE), room 176
Capabilities
• Prepares consistent, high-quality semi-thin and ultrathin sections for transmission electron microscopy and light microscopy.
Instrument Features
• Thickness feed range from 1 nm to 15 µm.
• Cutting speed range from 0.05 to 100 mm/s.
• Eucentric movement allows for optimal knife approach to the sample.
• Outstanding LED illumination (top, back, and transmitted light; independent control of the illumination modes) yields outstanding visibility of the knife edge and block face.
• Fully motorized knife stage and auto-trim function.
• Vibration-decoupled gravity-stroke cutting arm for chatter-free sectioning.
• Touchscreen control unit.