VCU Massey Cancer Center

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The VCU Microscopy Core Laboratory (Director Tytus Bernas, Ph.D.) is located in a 3000 sq ft facility spread over several rooms on the 9th floor of Sanger Hall. It is a fee-for-service core supported by the director and three FTEs that provides the instrumentation and expertise to facilitate a comprehensive spectrum of imaging methods and techniques. Instrumentation and services include:

1) Electron microscopy (TEM, SEM) is supported with a Jeol JEM-1400 TEM equipped with Gatan UltraScan 4000SP CCD camera and tilting motorized stage (montaging and 3D), and a Zeiss EVO 50 XVP SEM equipped with SE, VPSE & BSD detectors, extended variable pressure (up to 750 Pa), Deben coolstage, a water vapor introduction kit, and a Shuttle & Find stage adapter for correlative confocal / SEM imaging;

2) Confocal laser scanning microscopy, supported by three systems: A Zeiss LSM 710 (inverted) equipped with a 32-channel spectral detector plus 2 side PMT detectors, a transmitted light detector, a Becker & Hickl Fluorescence Lifetime Imaging system with 2 hybrid GaAsP detectors (for FLIM / FCS), a motorized XY stage, a PeCon low profile stage incubator, Shuttle & Find stage adapter, and 5 lasers (pulsed 405 nm, multi-line Argon [458/488/514 nm], 561 nm green diode, 633 nm HeNe & a pulsed 440 nm. The system is set up for FRET, FRAP and FLIM analysis. The second system is a Zeiss LSM 700 (upright) equipped with 4 solid state lasers (405 nm, 488 nm, 555 nm, and 635 nm). The scan head has 2 confocal PMTs, a variable secondary dichroic beamsplitter and a transmitted light detector. The third system is a Zeiss LSM 880, inverted) equipped with spectral array detector (GaAsP), 2 single-channel PMT detectors, 32-element image scanning (Airy) detector (GaAsP), motorized piezo stage, hardware autofocus, stage incubator and 5 lasers (405 nm, multi-line Argon [458/488/514 nm], 561 nm green diode, 594 nm HeNe and 633 nm HeNe. The system is capable of high-resolution (image scanning), FCS and fast live-cell imaging.

3) Multi-photon laser scanning microscopy, supported by a Zeiss LSM 510 META NLO (fixed stage upright) equipped with a Spectra-Physics Broadband (710-990 nm) MaiTai Ti:sapphire laser and 3 visible lasers (multi-line Argon, 561 DPSS, HeNe 633), 2 non-descanned detectors, 3 descanned detectors (including a META detector for spectral scanning) a transmitted light detector, and a variety of objective lenses (dry, oil, multi-immersion, water immersion). A Luigs & Neumann 380 FM workstation (with slice chamber & in vivo bridges) and physiology equipment (stimulators, VP timing unit, A/D converters, multiclamp amplifier, line heaters, peristaltic pump, pipette puller) are linked with the system to facilitate simultaneous imaging & physiological recording;

4) Live cell confocal microscopy supported by a Zeiss Cell Observer SD spinning disc confocal microscope, equipped with a Yokogawa CSU-X1A spinning disc, 2 Photometrics Evolve 512 cameras, an Axiocam MRm camera, a high resolution piezo driven Z stage, 4 lasers (405 nm, 458/488/514 nm, 561 nm, 635 nm), DIC, phase contrast, FRAP/uncaging and a stage incubation system (to support live cell / live tissue imaging, with regulation for temperature, humidity, CO2 and O2); 5) Total internal reflection (TIRF) microscopy, supported by an Olympus cellTIRF system equipped with 3 lasers (405 nm diode, 488 diode and 561 DPSS), AOTF, DIC, phase contrast, Andor iXon DU-897E EMCCD camera, a laser-based automated focus drift compensation system and a stage incubator (temperature, humidity, CO2); The system is set up for super-resolution imaging (PALM/STORM localization microscopy).

6) A Nikon N-SIM Structured Illumination Microscope. This system is equipped with 3 diffraction gratings, 4 lasers (405, 488, 561 & 640 nm), Andor iXon DU-897E EMCCD camera, a laser-based automated focus drift compensation system, motorized XY stage, a high resolution piezo-driven Z stage, motorized TIRF optics (with independent laser input), fluorescence optics, DIC, and a Tokai Hit stage incubator (to facilitate live cell imaging); The system is capable of super-resolution (SIM and localization), TIRF and combined SIM/TIRF imaging of live and fixed cells.

7) seven wide-field fluorescence microscopes, including: an Olympus BX51 configured for computer-assisted stereology and equipped with a Prior Proscan XYZ motorized stage, an Olympus DP72 CCD camera, fluorescence optics (DAPI / GFP / DsRed / Cy5), DIC, phase contrast, Visiopharm newCAST software and Olympus cellSens software. The second system is a Zeiss AxioImager Z2 automated microscope configured for morphometric analysis and equipped with a Q-Imaging CCD camera and a Zeiss Axiocam MRm, fluorescence optics, (DAPI / CFP / GFP / DsRed / Cy5), DIC, motorized XY stage, and Microbrightfield Neurolucida & StereoInvestigator imaging / image analysis software. The third system is a Zeiss Axiovert 200 equipped with a Hamamastu ORCA ER CCD camera, Colibri LED illumination unit (blue, green, red), fluorescence filter sets (DAPI, GFP, DsRed), white light LED and a stage heater. The fourth system is a Zeiss Axioimager A1 equipped with fluorescence optics (DAPI, GFP, DsRed), and an Axiocam MRc CCD camera. The fifth system is a Zeiss AxioObserver A1 equipped with fluorescence optics (DAPI, GFP, DsRed), and an Axiocam MRc5 CCD camera. The sixth system is a Nikon ECLIPSE E800M with fluorescence optics (DAPI, GFP, DsRed, Cy5), and a Diagnostic Instruments Spot RT CCD camera. The seventh system is a Zeiss Axiovert 40 equipped with fluorescence optics (DAPI, GFP, DsRed), and an Axiocam MRc CCD camera. A Zeiss Discovery V20 Stereo zoom microscope with transmitted, reflection and fluorescence illumination, 0.63x, 1.25x and 1.5x lenses, filter sets for GFP, CFP, YFP and DsRed and an Axiocam MRc CCD camera is available for micro/macroscopic imaging;

This equipment suite (supported by sputter coaters, ultramicrotomes, critical point dryers, etc.) allows the core to offer many types of imaging methods and services including: Sample preparation; Live-cell imaging; Immuno-localization (fluorescence, EM); Fluorescence recovery after photobleaching (FRAP); Fluorescence resonance energy transfer (FRET), Fluorescence lifetime imaging (FLIM), Fluorescence correlation spectroscopy (FCS). The core also provides multi-dimensional image analysis (image cytometry), restoration (including deconvolution) and visualization. A cell culture facility (with incubators and biological safety cabinet) is located on-site to support live cell imaging. Training is available for all of the imaging modalities and methods offered.