Microscopy and Imaging

The Microscopy and Imaging Laboratory provides DC CFAR investigators with priority access to instrumentation for microscopic image acquisition and data analysis, training on the use of available equipment, quality control of the acquired microscopic images and data analysis.

In addition to routine confocal imaging, the core offers its extensive infrastructure and expertise in life cell microscopic imaging and analysis and infrastructure for transmission electron microscopy.

Services and Instrumentation

• Zeiss LSM 510: The Zeiss LSM 510 confocal system is equipped with fully motorized Zeiss Observer Z1 inverted microscope, with 10x (Plan Neofluar, N.A. 0.3), 20x (Plan Apochromat, N.A. 0.8), 40x (EC Plan Neofluar N.A. 1.3), 60x (Plan Apochromat, N.A. 1.4, Oil, DIC) objectives, and 100x (Alpha, N.A. 1.46, Oil, DIC). Four lasers are available for excitation: (a) Argon 30 mW, emitting at 458, 477, 488, and 514 nm, (b) HeNe 1.2 mW emitting at 543 nm, (c) HeNe 5 mW emitting at 633 nm, and (d) Diode laser 30 mW emitting at 405 nm. The confocal system has three standard confocal photomultipliers and one transmitted light photomultiplier. The system is equipped with Zeiss integrated on-stage environmental chamber for control of CO2 and temperature for cell culture time-lapse experiments.

• Zeiss LSM 710: The Zeiss LSM 710 confocal system (NIH 1S10RR025565) on AXIO EXAMINER upright microscope and LSM 710 with a 34 spectral detection channels and two transmitted light photomultipliers, 5 laser lines - 458, 488, 514, 561 & 633 nm, control electronics, ZEN 2009 software and x/y/z motorized scanning stage. Additional 405 nm diode laser. Objectives: Plan-Apochromat 10x/0.45 D=2.0, Plan-Apochromat 20x/0.8 M27 D=0.55, W Plan-Apo 20x/1.0 DIC VIS-IR WD=1.8, Plan- Apochromat 63x/1.40 Oil DIC, Alpha Plan Apochromat 100x/1.46. On stage incubator (Tokai Hit, Japan) for long-term observations of explanted tissues and control of the temperature and environment either via perfusion or CO2 control.
• JEOL JEM-1200EX: The JEOL JEM-1200EX is a classic Transmission Electron Microscope (TEM) 100 kV equipped with 4 megapixel SIA-CAM.
• Ultramicrotome workstations: Reichert Jung Ultracut E, Sorval MT5000 and MT6000 are available for sectioning specimens for TEM imaging. A Thermo Scientific Microm HM650 V slicer is available for living tissue.
• Microscope Imaging Workstation: The microscope workstation has an Olympus BX-60 upright Fluorescence Microscope with a Ludl Focus Motor and an Evolution Digital CCD Camera connected to a computer with Image Pro Plus 6.2, including 3-D constructor module and ImageJ.
• Nikon Eclipse TE300: The Nikon Eclipse TE300 inverted microscope with Optronics CCD camera at 1MP resolution can perform phase contrast, bright-field and fluorescence imaging. It is also available equipped with a microinjection system.
• Image Analysis Workstation: The Image-Pro Plus v6.2 and Metamorph Image Analysis Workstation includes 3D constructor and deconvolution modules on Dell Precision 7400 computer with 64 Windows XP operating system. Velocity software on iMac 16 GB processor, supporting analysis of large 3D and 4D data is also available.
• Storm 860 Molecular Imager: The Storm 860 and personal densitometer with Image Quant software are available for blue and red lasers for excitation.

Leadership

The Microscopy and Imaging Laboratory is directed by Dr. Anastas Popratiloff, Director of the GW Center for Microscopy and Imaging Analysis.

Dr. Popratiloff is available to consult with DC CFAR members on experimental design, data acquisition approaches and data analysis strategies.