Confocal Facility

Dr. Vickery Trinkaus-Randall is a Professor of Biochemistry and Ophthalmology. She has successfully obtained NCRR shared instrumentation grants to fund the purchase of two of the instruments available to the Core. She is internationally recognized for her work on injury and remodeling of the cornea, and has extensive experience in novel microscopic techniques.

Two confocal microscopes were purchased under NIH NCRR grants and are housed in a laboratory in R918. 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 (*.xls) format.

A workstation with PC computer and 17 inch monitor includes freeware software for imaging including Image J, NIH Image and Retrac, in addition to the full proprietary software package offered by Zeiss, is available to all users.

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.

Access

The core facility will be available to any qualified researcher across BUMC. All persons will be trained on the instruments and qualified as independent after evaluation by staff. Abuse of the instrument will result in lack of access. Outside users will be allowed but priority will be given to BUMC users. There will be a cost differential for BUMC users (short and long term) versus external users. An Imaging course has been offered yearly in the Spring Semester and will be offered again in 08 ñ GMS CM-766.
Rates
The user fees are designed to help support the service contracts and the costs of supplies that are not supported by grants or institutional sources. The charges are commensurate with other universities in the Boston area. Pilot studies and new investigators collecting data for a grant application will be eligible for 4 hrs of free use and consultation. Cost for usage is proportional to time used.
The current rates for usage of the LSM510 confocals are:

BUMC Users:

Unassisted: $50/hr
Assisted: $75/hr
40 hrs:$1200
100 hrs:$2700
200 hrs:$4000
300 hrs/yr:$5000

External Users:

Unassisted:$100/hr
Assisted $140/hr
Rates for 40 hrs are $2400.

Training and teaching

One of the missions of the Core is to continually train users and this includes graduate students and postdoctoral fellows to enable them to enter and contribute to labs using the high end technologies that are a part of research. To this end, the Core teaches an Imaging Class yearly with hands on experience, lectures in graduate level courses, brings in speakers who are experts on imaging, and sends herself and staff to Quantitative Microscopy Courses held at either Mt Desert Island or Woods Hole to maintain the high level of expertise.