Much information for new users can be found in an entire section of this web site dedicated just to you.
Head on over to the New User Information section.

ALL CISR microscopes are available ONLY to those who have been trained in their use by a member of the RESOURCE STAFF. After training, you will be able to sign up and use the microscopes as needed. Detailed instructions on microscope use can be found on most Equipment pages.

CISR SERVICE PRICES [as of September 1st, 2018]

All CISR microscopes are in secured rooms, most requiring a VU ID badge for entry. CISR light microscopes are available for use 24/7. TEM and SEM are available Monday through Friday 9:00 a.m. – 6:00 p.m. with use after hours or on weekends limited to users with extensive experience who have undergone additional training.

Please remember that ALL scopes should be reserved before use through the online calendar reservation system.

For new experiments or for inexperienced researchers it is best to consult with CISR staff to insure that specimen preparation is optimized and to ensure you have a through understanding of microscope use. A brief meeting and/or training sesstion between the Resource staff and the researchers laboratory to iron out details can be arranged by contacting anyone on our Staff page. This meeting can help determine feasibility, provide suggestions for optimal use of the resource, and, if appropriate, alert the resource staff to unique details of specimen preparation for your project.

That depends on the sample, the fluorophore, and the information desired, but here are a couple of examples: a tissue staining protocol, and a cell staining protocol. Some notes on coverslips can be found here, and some information on anti-fade reagents and mounting media is located here.

Samples must be fixed prior to submission. Please see the Tips for Fixation Procedures for EM question on this page. If you have samples for immunolabeling or any other specialized protocol, please discuss your project with EM staff prior to sample preparation.

Fixation is typically achieved with a 2.5% glutaraldehyde solution in 0.1 M sodium cacodylate buffer. Fixation should be initiated at room temperature. Tissue should be fixed in a solution volume of at least ten times the sample volume. Specimen vials pre-filled with fixative solution are available for users in the refrigerator of the Processing Lab located in room T-3208 of MCN.

Larger volumes of fixative and buffer are available for whole-animal perfusion fixation (recommended). Please contact a member of our staff to arrange for pick up.

Samples for SEM must also be fixed. Typically these samples will be processed in the critical point dryer by EM staff and mounted for examination in the SEM.

To maintain ultrastructure, specimens must be fixed rapidly after the oxygen supply has been cut off. The specifics of fixation (fixative, concentration, time, temperature) and the buffering medium will vary for specific experiments. However, in general, fixation will involve buffered aldehydes which cross link proteins, carbohydrates, and to a certain extent lipids. The fixatives will cross link similar molecules that surround the sample (body fluids, cell culture medium, etc. ) so the sample should be washed briefly in a buffered solution before fixation.

Aldehyde fixatives penetrate tissue slowly, so samples must be small (less than 0.5 mm on two sides). For large specimens, mincing the sample directly in fixative after washing works best. This can be done by placing a drop of fixative on dental wax or parafilm and placing the excised and washed sample in the drop of fixative and then mincing with fresh razor blades. The EM staff can show you how this is done.

Cold depolymerizes cytoskeletal elements, so whenever possible, buffer washes, fixatives, and sample should be at room temperature or warmer to initiate fixation. For temperature sensitive samples, such as enzyme cytochemistry, the samples can be briefly warmed placed in fixative and then, after a minute, re-cooled. To store fixed samples for more than a few hours, the specimen should be placed on ice or in the refrigerator after fixation. For storage of samples more than a few days before processing, the fixative should be replaced with buffer after 24 hours.

All samples must now go through the EM online laboratory information management system located here. Please contact a member of our staff if you experience any problems or need access to the system.

Please note that specimens for EM processing should be fixed prior to submission and kept refrigerated until submitted. EM fixative and wash buffer can be obtained from the refrigerator in the Processing Lab located at T-3208 MCN.

CISRstore is a very large, very fast disk storage system available to all registered Cell Imaging Shared Resource users, and is the core's preferred location for storing image files. Click Here to view instructions on accessing the CISRstore.

The free LSM Image browser is here.

The free NIS-Elements Viewer is here.

The free Olympus Viewer is here.

The free ZEN browser is here.

Answer: Just ask!

CISR staff members are available to assist you with FRET, FRAP, time lapse, image tiling, and many, many other specialized procedures. Extended help may be subject to additional charges, but most answers are free. We are always ready to help.