The MIC operates in three different areas: equipment maintenance, user support, and data analysis. Equipment acquisition, maintenance and upgrade The facility operates three modern confocal microscopes, each optimized for certain applications: 1) A Zeiss LSM 710 inverted for high-resolution confocal imaging of fixed specimen and live cells. 2) A Zeiss LSM 780 for challenging specimen requirying both high resolution and high sensitivity. 3) A Nikon Spinning Disk / Total Internal Reflection Fluorescence (TIRF) hybrid microscope for high-speed confocal imaging or selective recording of membrane-bound events in live cells (TIRF). An 11-year-old Zeiss LSM 510 2-photon instrument was decommisioned recently due to severe degradation in the performance of the pulsed 2-photon laser. Replacement of this microscope for live tissue slices and live animal imaging is being discussed with the Office of the Scientific Director. The MIC also owns several high-end conventional (wide-field) light microscopes: 1) A fluorescence inverted Olympus IX8 for imaging fixed and live specimen, equipped with a Flash-4 high-speed camera. 2) An upright fluorescence Olympus BX61 for fixed slides. 3) A Zeiss Lumar fluorescence stereo-microscope with a color camera for large specimen, including dissection work. 4) A Zeiss inverted Axiovert equipped with a high-end Color camera for transmission imaging of visible stains and pathology specimens. Live imaging is supported on all confocal microscopes and on the Olympus IX81 with a stage-top chamber providing temperature, CO2, and humidity control. All confocals are equipped with a motorized stage that allows multi-location and tiled imaging experiments, and also an auto-focus device for long-term recordings (24-72 hours). In order to keep downtime to a minimum, Dr. Schram carries an NIH-provided cell phone and can be reached quickly, including outside business hours. For problems that require extensive repairs, the three confocal microscopes are covered by a manufacturers service contract and are usually serviced within 1 to 2 business day. User training and support After an initial orientation where their project is dicussed, users receives hands-on training on the light microscope best suited to their goals. The training covers the principles of fluorescence microscopy, confocal imaging, and optimum operation of the hardware and software involved. It is followed by periodic refreshers at the user's request, or when the staff feels the equipment is not being used optimally. Beyond this first and essential level, the facility provides training and support in other meaningful ways: whenever possible, Dr. Schram offers on-site assistance on equipment owned by individual investigators. Yearly training classes on light and confocal microscopy are organized by the facility. This initiative has been well received by the community in PNRC, especially the hands-on workshop on ImageJ organized by Dr. Schram. The MIC website (mic.nichd.nih.gov) is an important resource for tutorials and protocols for both fixed and live cell microscopy. Last but not least, the facility organizes on-campus demonstrations of new instruments and software by vendors such as Zeiss, Olympus, Photometrics, Nikon and Leica, where users can evaluate first-hand new equipment and technologies. During the past year, three such demonstrations were organized (GE Healthcare / DetaVision Elite, Zeiss / LSM 880 with Airy detector, and Nikon / A1R MP+ multiphoton). Data analysis The facility operates three high-end workstations running advanced image analysis software (Metamorph, Nikon Elements, Volocity, Imaris, Zeiss AIM and Zen). At the user's request, we provide training and support for each software package and, when required, write protocols or custom macros for high-volume image processing. The facility also offers extensive data storage with a 20-terabyte file server available for data transfer and long-term image storage. Facility usage As of this writing, the MIC has a total of 177 registered users in 58 laboratories. Following the recent decision to waive usage charges (Spring 2015), both the overall usage and the ratio of PNRC users not affiliated with NICHD is rising. However, usage has not reached a saturation point because users with basic imaging needs are oriented toward the older confocal microscopes onwed by NINDS (Dr. Smith is likewise steering advanced projects toward some NICHD confocal microscopes). Publications Since its inception in 2004, the work carried out in the MIC has resulted in more than 160 publications. For a complete list, head to: https://science.nichd.nih.gov/confluence/display/mic/Publications
Showing the most recent 10 out of 23 publications
