Quantitative Receptor
Autoradiography Services

Consultation
Users are advised to fill out a quantitative receptor autoradiography worksheet for their intended experiment. This will be reviewed by Dr. Richfield for discussion with you regarding appropriate sample sizes, experimental details, efficiency, feasibility, time frame, and cost.

Ligand Purchase
A large number of isotopically labeled ligands are commercially available to label receptors and transporters. Assays exist for studying those receptors and transporters in tissue sections using autoradiography. Ligands can be purchased through the Molecular Histology Center or by an investigator and shipped to us. The choice of a ligand needs to be made carefully and the Molecular Histology Center can assist with the choice of ligand and the isotope used.

Tissue Acquisition
Specific organs from defined mouse or rat strains can be obtained. Fresh frozen tissue is always used for quantitative receptor autoradiography to ensure the maximal number of transporters or receptors in the native state available for binding.

Tissue Sectioning
Tissue sections can be prepared for assays. Typical thickness is 10 microns. These can be cut and mounted onto pig gel subbed slides, dried, and stored at -70°C until used in assays. Any plane of orientation can be cut and multiple organs can be mounted to a single slide.

Experimental Assays
A variety of specific assays can be performed for studying receptors and/or transporters. Equilibrium studies can be performed to determine when equilibrium binding is achieved. Kinetic studies can be performed. Saturation studies to determine both the Bmax and KD of a receptor can be determined. Tissue distribution studies for a particular receptor or transporter can be done and comparisons can be made between different experimental conditions.

Film or Phosphor Screen Exposure
Users can choose either a permanent hard copy of the experiment using isotope-sensitive film or a digital image using a phosphor screen or both. Images from the phosphor screen can be downloaded from our website and viewed, manipulated, measured or exportted in other formats on your computer. Software for this can be downloaded from our web site. Films can be shipped directly to you.

Densitometry
Calibrated standards are always included with quantitative receptor autoradiographic exposures. The standards are calibrated to both 3H and 35S. Quantitative densitometry using those standards can be obtained in organs or regions of interest within an organ and under different experimental conditions. Data are provided in a spreadsheet for downloading from our website. Remote user-assisted densitometry is also available.

Image Capture
Digital capture of images from film and/or phosphor screens can be obtained in gray-scale or pseudocolor. Image processing or production of a montage of images can be prepared. All captured images are available for download from our website.

Project Page
All projects have a unique password protected page on our website containing information pertaining to your project including a tissue log, ligand, receptor and/or transporter examined, data spreadsheets for specific experiments, image files, and billing invoices for download.

Molecular Histology Center Environmental and Occupational Health Sciences Institute, 170 Frelinghuysen Road, Piscataway, NJ 08854 Phone: 732-445-3729 For additional information contact ekr@eohsi.rutgers.edu

Updated on Friday, June 03, 2005