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Cellular and Molecular Imaging

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CELLULAR AND MOLECULAR IMAGING

 

     

Scanning serial block face microscope

VolumeScope is a Serial Block-face Imaging (SBFI) instrument for automated acquisition of large volumes. It integrates with Thermo Fisher Scientific SEM platform. The key enabling technologies, hardware, and software include:

  • Novel Field Emission Electron Optics for both high resolution and exceptional contrast, meeting all imaging needs at low beam currents.
  • Fully-integrated, compact, stage-mounted ultra-microtome for in-situ sectioning.
  • Enables direct correlation of images from any light microscope covering large volumes.
Nikon SoRa Spinning Disk Microscope
[Image source: Nikon]

The Nikon SoRa (super resolution by optical reassignment) provides conventional confocal scanning, SoRa at 2.5X and 4X resolution, and TIRF imaging captured by 3 hamamatsu cameras. It allows a 1.4X improvement in lateral resolution beyond the optical diffraction limit.

The Nikon SoRa can collect super resolution fluorescence images as well as conventional confocal, TIRF, wide field, and transmitted light images (bright field and DIC). The scope has 10x, 20x, 20X water, 25X silicon, 40X water, 60x TIRF oil, and 60x oil objectives. A water adjustment system allows for the proper water to be applied throughout the experiment while a stage-top incubator adjusts temperature and CO2. The iLas2 and photostimulation with galvo scanner provides 360 degree TIRF imaging.

Lasers: LunF-XL solid state laser combiner with 405, 445, 488, 514, 561, 594 and 640nm imaging lines and 405, 473 stimulation lines

Filters: DAPI, GFP, RFP, Cy5

Cameras: 3 Hamamatsu Back-Thinned Orca Fusion CMOS cameras

Software:  NIS Elements, 2D/3D deconvolution, iLas2

Computer: HP Z8 microscope computer and separate workstation for analysis

zeiss LSM 710

Uses a Zeiss AXIOExaminer upright microscope stand with transmitted (HAL), UV (HBO) and laser illumination sources. Can collect transmitted light images (bright field and DIC) as well as conventional and confocal fluorescence images. The scope has 10x, 20x, 40x (oil), and 63x (oil) objectives. Provides Z-series, 3D/rotatable reconstruction, time-lapse analysis for following cellular events in live samples, and co-localization of multiple proteins and multicolor imaging of fixed samples with a wide array of imaging applications including protein localization analysis, cytoskeletal studies, live cell imaging and cell cycling studies.

Fluorescence Filters: GFP, DsRed

Lasers:  488 / 555 / 594 / 633

Detectors: 34 channel confocal and transmitted light detectors

Software: ZEN software to control the microscope, scanning, laser module, tools and the image acquisition and processing

Computer: LSM710 is controlled via a HP Z800 workstation with the Windows 7 (64 bit) operating system

zeiss LSM 700

Includes inverted stage (4 laser lines – 405 / 488 / 555 / 647) equipped with a live cell imaging incubator and an automated stage for automatic scanning and tiling of large images.

Uses a Zeiss AXIOExaminer inverted microscope stand with transmitted (HAL), UV (HBO) and laser illumination sources. Can collect transmitted light images (bright field and DIC) as well as conventional and confocal fluorescence images. Has 10x, 20x, 40x (oil), and 63x (oil) objectives and a fully automated platform, including a live cell imaging incubator and automatic scanning and tiling of large images. Provides Z-series, 3D/rotatable reconstruction, time-lapse analysis for following cellular events in live samples, and co-localization of multiple proteins and multicolor imaging of fixed samples with a wide array of imaging applications including protein localization analysis, cytoskeletal studies, live cell imaging and cell cycling studies.

Fluorescence Filters: DAPI, DsRed, GFP, mPlum

Lasers:  488 / 555 / 594 / 633

Software: ZEN software to control the microscope, scanning, laser module, tools and the image acquisition and processing

Computer:  HP Z820 workstation with the Windows 7 (64 bit) operating system

Transmission Electron Microscope

The FEI Tecnai G2 Spirit Twin Transmission Electron Microscopy (EM) is a multi-purpose- high resolution instrument. The cooled column and LaB6 emitter povide stable low-dose observation of beam sensitive samples while the 20-120kV capacity allows for imaging biological and material science samples.  Features include:

  • An FEI Tecnai G2 Spirit Bio-Twin TEM operating at 20-120 kV
  • 0.2nm line resolution
  • LaB6 emitter
  • Gatan Rio CCD camera
Multiphoton Microscope
[Image source: Bruker]

Provides simultaneous imaging and electrophysiology, plus uncaging and photostimulation. Secondary scan path allows for simultaneous imaging and photostimulation. Point photoactivation achieved with  sub-micron positioning controls. 

Leica Stellaris 8 Confocal Microscope
[Image Source: Leica Microsystems]

STELLARIS 8 is a true confocal point scanning system, including a tuneable White Light Laser and a UV laser extending the detection range from 405 to near InfraRed (850).  The system comes with 4 highly sensitive prism-based spectral detectors with computer controlled adjustable emission bandwidth.  Lightning technology provides super resolution imaging down to 120nm resolution in xy.  Both Falcon and TauSense technology provide a set of tools based on fluorescence lifetime imaging microscopy (FLIM) for improved image quality and separation of spectrally overlapping fluorophores. The system has high resolution and high depth objectives to accommodate a wide range of research needs. 

Features include:

  • High-end HpZ6G4 workstation with CUDA support for high workloads
  • LAS X Stellaris Software for live data mode, c3D visualization, olocalization, FRAT, FRET, and electrophysiology
  • LAS X Lightning Expert is advanced software for confocal, STED, and multiphoton data
  • LAS C Co-Localization
  • Power HyD S detector – high performance
  • Power Hybrid Detector HyD X- specialized for FALCON (Fast Lifetime CONtrast)
  • Tuneable lasers up to 850nm
  • Objectives – 5x, 10x, 20x, 25x, 40x, 63x
  • Additional workstation with full software suite for offline data analysis

To initiate use of cellular and molecular imaging equipment:

  • Contact facility director Christie Lacy, (christie21@vtc.vt.edu).
  • Complete required equipment training with facility manager.
  • Access to the scheduling tool is granted upon completing of training.

Fees

(per hour for internal and external users)
  • Zeiss 700: $20 internal, $32 external
  • Zeiss 710: $25 internal, $43 external
  • Bruker/Zeiss 2-Photon: $40 internal, $69 external
  • FEI TecnaiSpirit TEM: $35 internal, $61 external
  • Nikon SoRa Spinning Disk: $25 internal, $43 external
  • Apreo Serial BlockFace EM: $20 internal, $70 external

 

  • Training is required for each instrument.
  • Users must begin by contacting facility director Christie Lacy, (christie21@vtc.vt.edu).
  • Training is scheduled and provided one-on-one by the facility director.
  • Each user must complete at least two training sessions.
  • The intial training session typically takes 2-3 hours and covers basic operating of the instrument. 
  • The second session is for the user to practice with their own sample. 
  • Any subsequent training is based on the needs of the user.
  • Users may schedule 3-hour time blocks each day on an instrument.
  • Users must be trained on an instrument to schedule use of it.
  • Currently some instruments have other restrictions on the amount of time that can be reserved.
  • Two weeks prior to the day of a reserveration, users may extend their block past three hours if time on the instrument is available.
  • Users more than 15 minutes late to a reserved time block may lose their time to another user.
  • Users must clean up after themselves.
  • Only leave the instrument on for the next user IF they are present have asked you to do so. 

All Cellular and Molecular Imaging equipment is located at the Fralin Biomedical Research Institute at VTC in Roanoke, Virginia.

Thermo Fisher Scientific Apreo Volumescope Scanning Serial Block Face Electron Microscope: Room 2306A, 4 Riverside Circle

Nikon SoRa Inverted Spinning Disk Confocal Microscope: Room 1036, 4 Riverside Circle

Zeiss 700 Confocal Microscope: Room R1028, 2 Riverside Circle

Zeiss 710 Confocal Microscope: Room R1028, 2 Riverside Circle

Transmission Electron Microscope with Cryo Capacity:  Room R2029, 2 Riverside Circle

Bruker Ultima in vitro Multiphoton Microscope: Room R1015, 2 Riverside Circle

Leica STELLARIS 8 Spectral Confocal Microscope: Room R3024, 2 Riverside Circle

Parking: During business hours, visitors may park in designated visitor parking places at the entrance to the parking garage at 2 Riverside Circle. Visitors may also park on the 2nd - 6th floors of the parking garage located at 5 Riverside Circle. On nights and weekends, visitors may park anywhere in the 5 Riverside parking garage, as well as the surface lot and parking deck. All parking is free.

Contacts

Facility Director
Christie Lacy
christie21@vtc.vt.edu

Facility Faculty Director
Shannon Farris, Ph.D.
farrissl@vt.edu