Advanced Bioimaging Laboratory

Contact Info:

Director Kirk Czymmek, PhD

Pricing:

Available Upon Request

Summary:

The Advanced Bioimaging Laboratory is a 1,500 sq/ft suite of powerful imaging and analysis platforms as well as supporting preparation equipment for routine and advanced plant microscopy applications, including confocal, super-resolution, macro microscopy, wide-field fluorescence, dissecting,and transmission electron microscopy. The facility has the following capabilities:
1. Generate high-resolution 3D and 4D image data
2. Produce large area maps and sophisticated multi-location and analysis routines
3. Perform brightfield, phase contrast, DIC, sensitive imaging of a variety of fluorphores including time-lapse fluorescence imaging
4. Screen for fluorescent protein expression and white light imaging of plant morphology
5. 3D visualization and analysis with deconvolution optical sectioning
The facility also includes equipment for cryo-preparation, processing frozen samples, fast-freeze substitution, ultrathin sectioning, serial sectioning, microdissection, handling frozen sections to glass slides, and sample preparation.

Equipment:

Leica SP8-X confocal microscope on an inverted platform
- 405 nm diode laser
- Argon lasers: 458 nm, 476 nm, 488 nm, 496 nm, 514 nm
- Detectors: photomultiplier tube (PMT), supersensitive hybrid detectors (HyD), transmitted light detectors (PMT)
- Motorized microscope stage with Super Z-galvo scanning insert for fast and precise Z movement, standard insert for microscopy slides and 3 – 6-cm glass-bottom Petri dishes; 96-well plate stage insert
- Objectives:
  - Correction Collar 0.14-0.18
  - HC PL APO 100x/ NA 1.44 OIL
  - HC PL APO 10x/NA 0.40 DRY
  - HC PL APO 20x/NA 0.70 DRY
  - HC PL APO 40x/NA 1.10 WATER CS2
  - HC PL APO 63x/NA 1.20
- WLL Whight light laser (allows as many as 200 laser lines starting from 470 nm to 750 nm)
Nikon Ni-E wide-field fluorescence imager with an upright configuration
- 16-bit monochromatic digital camera (Hamamatsu Orca Flash 4.0)
- Fluorescence filter cubes:
  - CFP Exc 437/20x Em 480-40 nm Dichroic T455lp 
  - Cy5 Exc 620/60 Em700/75 Dichroic T660lpxr
  - DAPI Exc 350/50 Em 460/50 Dichroic 40DCLP
  - dsRed (TRITC/Cy3) Exc 545/305 Em 620/60 Dichroic T570lp 
  - EGFP Exc 470/40 Em 525/50 Dichroic 495
  - YFP Exc 500/20x Em 535/30 nm Dichroic T515lp 
- Leica CM 1950 Cryostat CryoJane system
- Leica EM ICE High Pressure Freezer
- Leica Ultracut UCT Ultramicrotome
- Objectives:
  - Nikon Plan Apo 10x/0.45 DRY ∞/0.17 WD 4.0
  - Nikon Plan Apo 20x/0.75 DRY ∞/0.17 WD 1.0
  - Nikon Plan Apo 60x/1.2 WATER WD 031-0.28
  - Nikon Plan Apo 60x/1.4 OIL Ph3 DM ∞/0.17 WD 0.21
- Software: ORS Dragonfly, Stereology Analyzer v4.3.3, Zeiss Zen Black SR, Zeiss Zen Blue, SVI Huygens Essential Deconvolution
- The Thermo Scientific Helios 5 Hydra DualBeam plasma focused ion beam Scanning Electron Microscope (PFIB-SEM)
- Thermo Scientific Talos 120C G2 Transmission Electron Microscope
- Vibratome 1000 Sectioning System
Super-Resolution Zeiss Elyra 7 microscope on an inverted platform
- DuoLink for parallel imaging on two cameras: 2x pco.edge sCMOS (version 4.2 CL HS) 
- Lasers: 405 nm, 488 nm, 561 nm, 642 nm
- Lattice SIM2, Apotome SIM2, TIRF, 2D & 3D Single Molecule Localization Microscopy capabilities
- Objectives:
  - alpha Plan-Apochromat 100x/ NA 1.46 OIL DIC M27
  - C-Apochromat 40x/NA 1.2 Water Corr FCS; C-Apochromat 63x/NA 1.2 Water Corr M27
  - EC Plan-Neofluar 10x/NA 0.3 M27 DRY
  - Plan-Apochromat 40x/NA 1.4 Oil DIC M27
  - Plan-Apochromat 63x/NA 1.4 Oil DIC M27
Zeiss AxioZoom.V16 microscope
- Camera: ZEISS Axiocam 512 color – color CCD camera 12 Megapixel resolution @ 4250 x 2838, 3.1 x 3.1 um pixels @ 14 bits/pixel, full resolution live image @ 10 fps, max live image speed 36 fps
- Fluorescence filter cubes:
  - CFP Filter set 47 (Ex: BP 436/25; BS FT 455; Em: BP 480/40) 
  - Cy5 Filter set 50 (Ex: BP 640/30; BS FT 660; Em: BP 690/50) 
  - DAPI Filter set 49 (Ex: G 365; BS FT 395; Em: BP 445/50) 
  - dsRed Filter set 43 (Ex: BP 545/25; BS FT 570; Em: BP 605/70) 
  - eGFP Filter set 38 HE (Ex: BP 470/40; BS FT 495; Em: BP 525/50) 
  - YFP Filter set 46 (Ex: BP 500/25; BS FT 515; Em: BP 535/350) 
- Lumen Dynamics X-Cite XYLIS LED Fluorescence Microscope Light Source; Excitation of commonly used fluorophores such as DAPI, CFP, GFP/FITC, YFP, TRITC/mCherry/Texas Red, Cy5 and Cy7
- Objectives:
  - PlanNeoFluar Z 1.0x/0.25
  - PlanNeoFluar Z 2.3x/0.57

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