Two confocal microscopes are housed in a laboratory in R918 that were purchased from NIH NCRR grants.
The confocal systems include a Zeiss Axiovert 100M microscope with argon laser lines (458 and 488 nm) and 2 HeNe lasers (568 and 633 nm), LSM 510 electronics control module v1.6 with 3 confocal detection channels for fluorescence or reflected light, and 10,20,25,40 and 63x objectives with DIC sliders for 25 and 40x objectives. Images are captured on a computer with command driven proprietary Zeiss LSM software. Standard software includes system configuration, ReUse, Acquisition modes, Auto-Z, crop, ROI scan and bleach, FRET, multitracking to minimize problems of cross-over, image display, analysis, processing and archiving.
The physiology and time series programs are present. Expert users are permitted usage of a flow through system in collaboration with Dr. Trinkaus-Randall. In April 2006 a second LSM 510 Zeiss Axiovert 200M (v4.0) with 5 argon laser lines, 2 HeNe lasers (568 and 633 nm) and a 405 nm laser was added. This allows people to use DAPI, Hoechst, ethidium bromide, Alexas of multiple wavelengths, GFP and numerous other dyes. Standard software includes system configuration, ReUse, Acquisition modes, Auto-Z, crop, ROI scan and bleach, FRET, multitracking to minimize problems of cross-over, image display, analysis, processing and archiving.
Programs include the physiology, time series using multiple regions, tileware, autofocus and stitchart. The objectives include a 10x dry, 20x dry, 2- 40x (water and oil), and a 63x oil objective. The instrument is equipped with a motorized stage and environmental chamber. Live cell imaging can be performed at 768 msec /frame on both instruments in a 512 x 512 configuration. Files are saved in the data base format and can be exported using a large number of formats and movies can also be exported. Using the Zeiss Software, one can assess intensity values (ROI), 3-D images, histograms throughout structures, co-localization. This data can be saved in Excel and exported. A workstation with PC computer and 17” monitor includes freeware software for imaging including Image J, NIH Image and Retrac in addition to the full proprietary software of Zeiss that is available to all users.
Additional Instruments – Additional instruments include workstations for the confocals, incubators for housing samples, dissecting microscopes, 1 Upright Leica Microscope with Epifluoresence and 1 inverted Nikon Microscope with a SPOT for capturing images.