Services

The WUCCI provides a complete service pipeline for light, electron, ion and 3D X-ray microscopy which is comprised of the following elements:

  • Affordable access to state-of-the-art light, electron, ion and 3D X-ray microscopy
  • Expert support in assay design, training on instrumentation and assistance with data acquisition
  • Analysis of multi-dimensional imaging datasets using commercial or custom-written algorithms

The WUCCI provides access to the following services: 

1. Light Microscopy

  1. Routine Microscopy:  Both upright and inverted microscope platforms capable of both Differential Interference Contrast (DIC) and multi-color fluorescence (DAPI/FITC/TRITC/Cy5) of fixed samples.
  2. Slide Scanning Microscopy:  An automated slide-scanning platform capable of scanning complete slides in either bright-field or multi-color fluorescence (DAPI/FITC/TRITC/Cy5) modes. System is equipped with a slide autoloader and automation software.
  3. Confocal Microscopy:  An upright confocal microscope platform capable of both transmitted light Differential Interference Contrast (DIC) and multi-color fluorescence imaging of fixed mounted cell cultures or tissue sections as well as living specimens such as C. elegans and zebrafish model systems. The system is also equipped with a second scanner for photo-bleaching experiments and photo-stimulation paradigms such as optogenetics.
  4. Live-cell Microscopy:  Two inverted confocal microscope platforms for 3D imaging of living specimens equipped with live-cell incubation chambers capable of controlling CO2, O2, humidity and temperature. The first is a resonant scanning LSM equipped with six laser lines (405, 445, 480, 515, 561 and 640nm) capable of spectral and FRET imaging, the second is a spinning disk confocal equipped with four laser lines (405, 480, 561 and 640nm) and two sCMOS cameras capable of simultaneous two-color imaging.
  5. TIRF / STORM Microscopy: An inverted platform equipped with a with live-cell incubation chambers\ capable of controlling CO2, O2, humidity and temperature and a motorized TIRF illuminator with four laser times (405, 480, 561 and 640nm) and two cameras. One camera (sCMOS) is designated for high-speed TIRF imaging, the other camera (EM-CCD) is designated for single-molecule imaging, specifically STOchastic Reconstruction Microscopy which enable imaging beyond the diffraction limit of light at ~20-40nm resolution in XY and 100nm in Z.
  6. SIM Super-Resolution Microscopy: An inverted platform equipped with four laser times (405, 480, 561 and 640nm) and two EM-CCD cameras capable of simultaneous two-color imaging at twice the resolution of confocal microscopy of fixed mounted cell cultures and thin tissue sections.
  7. Two-Photon Microscopy: An inverted two-photon microscope capable of multi-color fluorescence imaging of fixed mounted cell cultures or tissue sections as well as live specimens such as in vivo mouse imaging. The system is equipped with a Coherent Discovery laser for two-photon excitation and Gallium Arsenide Phosphide (GaAsP) detectors for high-sensitivity detection of fluorescence signals.

2. Electron / Ion Microscopy

  1. Transmission EM:  A JEOL 1400 Transmission Electron Microscope (TEM) boast high resolution and high contrast imaging on an easy to use automated platform. The system features 0.38nm point resolution and is configured for imaging at either 80kV or 120kV. It has a 5-axis precision Piezo motorized stage for large area montages and can accommodate tiel angles up to +/- 10 degrees for analglyph production. The system is equipped with an AMT XR111 high-speed 4k x 2k pixel phosphor scintillated 12-bit CCD camera. 
  2. Scanning EM: A Zeiss Merlin Scanning Electron Microscope (SEM) capable of analytical imaging of whole samples. Equipped with a Gemini II column, this SEM has superb low kV performance, and the ability to fine-tune lens parameters for optimization of image quality and acquisition speed. The voltage can be adjusted from 0.02-30kV (10pA to 40nA probe current) with 3.0nm resolution at 0.2kV, 1.8nm resolution at 1kV and 0.9nm resolution at 15kV.
  3. Focussed Ion Beam-SEM: A Zeiss Crossbeam 540 Focussed Ion Beam-Scanning Electron Microscope (FIB-SEM) capable of analytical imaging of whole samples and 3D nanotomography of embedded samples. Equipped with a Gemini II column, this SEM has superb low kV performance, and the ability to fine-tune lens parameters for optimization of image quality and acquisition speed. The voltage can be adjusted from 0.02-30kV (10pA to 40nA probe current) with 3.0nm resolution at 0.2kV, 1.8nm resolution at 1kV and 0.9nm resolution at 15kV. The Crossbeam 540 is also equipped with a Capella Ga-Liquid Meta Ion Source (Ga-LMIS) column that is capable of 3nm milling resolution at 30kV. The column can be set to 0.5-30kV (1pA to 100nA probe current). When coupled with the unique nature of the GEMINI II column imaging can be performed using only the SEM column, only the FIB column, or simultaneous SEM imaging / FIB imaging or milling for easy monitoring of routines. For masking and charge compensation purposes, the instrument is equipped with two Orsay-Physics gas injection systems (GIS) with both Pt and C precursor gases. In addition to the included imaging software, the Crossbeam 540 is equipped with the ATLAS 5.0 scan engineer with boasts gigapixel imaging capabilities (50k x 40k pixels) for large-area imaging projects.

3. X-ray Microscopy

  1. X-ray Microscopy:  A Zeiss Xradia 520 Versa X-ray microscope is available on a collaborative basis for the sub-micron tomographic imaging of tissues samples from microns to many centimeters in size. 

4. Sample Preparation

  1. Routine tissue EM:  Service includes processing and embedding (maximum of 6 blocks), cutting 1 micron thick plastic sections stained with toluidine blue, cutting of thin sections on standard 200 mesh grids, post-staining in uranyl acetate and lead citrate, and consultation. 
  2. Tissue culture EM:  Service includes routine procedures described above but requires additional specimen preparation and cutting time.
  3. Immuno-EM:  Services include LR White embedding media or pre-embed immunostaining, processing and embedding (maximum of 6 blocks) or cutting 1 micron plastic sections on slides - stained for toluidine blue.  Additional services include cutting thin sections on grids, primary antibody staining (primary antibody supplied by investigator), secondary antibody gold labeling (secondary antibody charged to investigator), counterstaining with OsO4, uranyl acetate, lead citrate, and consultation.
  4. Deep-Etch-EM: Service includes slam freezing of samples in liquid helium, freeze etching and free fracture as well as platinum replica production onto TEM grids.
  5. Negative Staining: Services include preparation of formvar-coated grids and negative staining techniques appropriate for the virus or particle to be visualized.
  6. Metal coating: Service includes critical point drying of samples and subsequent metal coating for topographic imaging and 3DEM in the SEM and FIB-SEM microscope platforms.