The CEMC offers training for and access to the following instruments: Transmission Electron Microscope, Ultrathin Microtome, Vibratome, Olympus Confocal Microscope, Zeiss Confocal Microscope, and Image Analysis Workstation. The CEMC also has equipment marked with (*) for emerging technology development and currently only operated by CEMC staff.
FEI Tecnai G2 12 Transmission Electron Microscope (TEM) with a Gatan OneView 16 Megapixel Camera is a TEM with an operating voltage range of 20 to 120 KV. Electron micrographs can be collected with this microscope with magnification of thin samples (<200 nm) up to 700,000 times. A Gatan OneView 16 Megapixel Camera is for digital image acquisition with imaging and in-situ data capture at full 4k × 4k resolution.
Olympus FV1000 Confocal Microscope is featured the world’s first twin scan system. This system was upgraded with a 748nm laser detector for CY7. It is upright model and is able to analyze objects down to a 2 nm wavelength resolution. Image size could be up to 4096 × 4096 – pixels.
Zeiss LSM880 Airyscan/CY7.5 Confocal Microscope offers high sensitivity, enhanced resolution in x,y and z, and high image-acquisition speed in one system. Users achieve a 1.7× higher resolution in all spatial dimensions, 140 nm laterally and 400 nm axially. The improved sensitivity leads to better image quality and increased speed. The whole imaging process is possible with standard sample preparation and labeling protocols. With Airyscan, ZEISS introduces a new concept. Instead of detecting signals with a single point detector, a multichannel area detector with 32 elements collects all the light from an Airy disk simultaneously. Hereby, each detector element functions as a single, very small pinhole. Knowing the beam path and the spatial distribution of each Airy disk enables very light-efficient imaging: researchers can now use all photons that the objective collected.
* Zeiss Sigma VP Field Emission Scanning Electron Microscope (SEM) with Gatan 3View2 XP System is used for image collection for 3-D reconstruction of neurons. The 3-D SEM system has the follow properties: (a) ultrathin microtome inside the scanning electron microscope chamber allows repeated cutting and imaging; (b) cut thickness can be set at 30 nm, even less than 15 nm; (c) the maximum sample size is about 1 mm × 1 mm × 0.6 mm; (d) the efficiency is very high. A 100-slice dataset can be collected in a few hours. A 1000-slice dataset can be collected with an overnight run.
Equipment for Tissue Sectioning – Leica VT1000S Vibratome, Leica UC-7 Ultrathin Microtome, and Leica Glass Knife Maker. Leica VT1000S Vibratome is used to section the whole brain in 30-200 µm thickness. Leica UC-7 Ultrathin Microtome is used to section samples in resin block in 60-70 nm thickness. Leica Glass Knife Maker is used to make glass knives for ultrathin sectioning before changing to a diamond knife.
Equipment for Tissue Processing – Blue M Regular Oven, Lindberg Blue M Vacuum Oven, Nikon SMZ150 Dissecting Microscope, and Leica EM AFS2. Blue M Regular Oven is used for pre-embedding resin polymerization. Lindberg Blue M Vacuum Oven is used for post-embedding resin polymerization. Nikon SMZ150 Dissecting Microscope is used to select the ROI of samples. Leica EM AFS2 performs freeze substitution and progressive lowering of temperature techniques and allows low temperature embedding and polymerization of resins.
* Equipment for emerging technology development in Correlative Light and Electron Microscopy (CELM) – RMC Automatic Tape Collecting Ultramicrotome (ATUMtome), Zeiss OxioObserver Microscope, Leica ACE600 Sputtering and Carbon Coater, and Ted Pella easiGlow Glow Discharge System. ATUMtome is used to collect hundreds to thousands of serial ultrathin sections onto to a tape substrate for later analysis and SEM imaging. Zeiss AxioObserver Microscope with ZEN Blue software is used to collect fluorescent images. Leica ACE600 Sputtering and Carbon Coater is used to produce thin, but strong support layers for SEM high-resolution imaging of structured non-conductive specimens. PELCO easiGlow™ Glow Discharge Cleaning System is primarily designed for cleaning TEM grids and preventing wrinkles in the polymer tape in CLEM.
Image Analysis Workstation – Available software: Imaris, Amira, Dragonfly, Image J, Image Pro, Huygens Professional for deconvolution, Zen Lite, Adobe, etc.