UMMC Electron Microscopy Facility

Facility Description

The UMMC electron microscopy facility is located in the Department of Biochemistry of the University of Mississippi Medical Center. The facility specializes in imaging of multiprotein and protein/nucleic acid complexes and is designed for acquisition of images of both negatively stained and frozen hydrated samples. These images can then be utilized in computational two and three dimensional image processing analyses.

The imaging part of the facility consists of: a LEO 912 AB transmission electron microscope with Koehler illumination, low dose imaging, an integrated OMEGA energy filter for elemental analysis and a liquid nitrogen-cooled cryostage ; all tools and transfer equipment for cryospecimen preparation and transfer; and a dedicated darkroom suite.

The computational analysis equipment includes: a 2500 X 2500 dpi AGFA scanner for digitizing negatives; a Silicon Graphics Octane workstation and with porting to a 64 processor Origin for parallel applications; and a Kodak dye sublimation printer for digital images.

The facility is under the direction of Dr. Mona T. Norcum to whom inquiries should be addressed. Contact information is as follows:

Mail :
     Biochemistry Department
     University of Mississippi Medical Center,
     2500 North State Street, Jackson, MS 39216-4505

Phone: 601-984-1527
FAX: 601-984-1501

Email: mnorcum@biochem.umsmed.edu

Guidelines for facility use

A. Facility use and acknowledgement

Use of the facility can be arranged as either collaborative research or on a fee-for-service basis. Regardless of basis, all publications that contain images obtained at the facility or data derived from such images must reference the UMMC Electron Microscopy Facility. Collaborative research assumes co-authorship of such manuscripts or abstracts.

B. Fees

Standard cost rates of fee-for-service work are:
     Sample preparation: $120 set-up charge plus $75 per vitrified sample or $40 per negatively stained sample
     Microscope time: $75 per hour
     Data documentation:
          Film negatives: $2.25 each
          Digitization with data storage on CD-ROM: $11.50 per negative
Note: Rates are higher for outside investigators due to added UMMC indirect costs.

Advanced data analysis (image averaging, three-dimensional image reconstruction) are not considered a standard part of the facility services. Please discuss these items with the facility director.

C. Billing

All billing and payment will be through the UMMC Accounting Office on behalf of the UMMC Electron Microscopy Facility. After approval of the proposed project by the facility director, a Purchase Order Number must be provided before work begins.

D. Project proposals

All requests for use of the facility should be made to the facility director and must contain the following information:

1. Rationale for the study

In order to judge feasibility of the proposed work, all potential users must provide a brief description of the overall goal of the project with specific details of the information desired from a the study. For example, a project may be designed to give a simple description of a particle’s size and shape, may involve mapping of component topography with antibodies or other labels, or may aim for reconstruction of a three-dimensional structure. The results of any prior imaging studies should be referenced.

2. Sample description

The relevant biochemical properties of the sample must be provided, e.g. type of material (protein, nucleic acid, viral particles, etc.). For single particle imaging, useful information includes the particle molecular mass, temperature stability, required buffer composition, etc. If any sample manipulations must be done at the facility beyond grid preparation, such as labeling or additional purification, all details must be included.

3. Hazardous materials

Any samples that are radioactive, contain live viruses, or otherwise fall under NIH biohazard guidelines must be fully identified. The facility staff reserves the right to refuse to work with such materials.

Guidelines for samples

A. Appropriate samples

The minimum particle size for routine structural analysis is approximately 400 kDa. Depending on the particle size, final sample concentrations are typically 15 - 100 micrograms per milliliter and as little as 50 - 100 microliters may be needed. However, in order to determine the optimum range, it is best to provide at least 50 -100 microliters at a concentration 100 - 200 micrograms per milliliter.

B. Appropriate buffers

Most common biological buffers are amenable to preparation of stained or frozen samples. However, high salt and high glycerol concentrations are problematic.

The University of Mississippi offers equal opportunity in employment and education, M/F/D/V.
Copyright © University of Mississippi Medical Center Department of Biochemistry
This document was last modified on
Page design by Anthony.