SAMPLE COVER LETTER
MUST BE ON UNIVERSITY LETTERHEAD

August 12, 1994

Mr. Donald L. Munroe
Radiation Control Officer
212 Nuclear Sciences Center
P O Box 118340

Dear Mr. Munroe:

Attached is my request for approval to use radioactive isotopes in my research program at the University of Florida.

I have used radioactive isotopes for approximately 10 years while at the Virginia Polytechnic Institute and State University.

For historical information, I have also attached my approved isotope authorization forms for my last renewal application at VPI and SU. I have indicated in red ink on the VPI and US forms the personnel who moved with me to the University of Florida.

All of our critical experiments require the use of radioactive isotopes. Therefore, we are very anxious to receive approval to use isotopes at the University.

Sincerely yours,

John Smith, Ph.D.

Professor

SAMPLE PROPOSAL

DATE:

TO:         Radiation Control Committee

ATTN:     Donald L. Munroe, Radiation Control Officer
                Box 118340

FROM:    Dr. John Smith (Principal Investigator)
                Professor
                Department of Microbiology and Cell Science

SUBJECT: Proposal to use C¹⁴, H³, P³³, P³², S³⁵, and I¹²⁵ (inorganic and organic forms) in tracer labeling studies in microorganisms, plant cells, and in biochemical reactions

1. Brief Description of Studies:
   Radioactive isotopes will be used in three different types of studies:

Radioactive organic and inorganic compounds will be used to measure the activities of different types of enzymes in test-tube assays. These radioactive assays will be used to quantify levels of enzymes in crude cell extracts and in purified enzyme preparations. In addition, substrates labeled in specific positions will be used in enzyme mechanisms studies. The types of radioactive substrates which will be used include C¹⁴-labeled bicarbonate; C¹⁴, P³², H³-labeled nucleotides and sugars, S³⁵-labeled amino acids and proteins, etc.

Radioactive iodine (I¹²⁵) will be covalently linked to enzymes (and other proteins) and antibodies. The resulting radioiodinated proteins will be used in protein/antibody binding measurements in test tube assays and in sucrose density-gradient centrifugation assays.

Radioactive inorganic and organic compounds will be added to the culture medium of plant cells, algae, and microorganisms to label intracellular metabolites, proteins, and nucleic acids. The cells will be broken and the distribution of radioactive label in the various intracellular metabolites and macromolecules will be ascertained after their fractionation by biochemical procedures. The types of radioactive compounds added to the culture medium will include C¹⁴-labeled bicarbonate, glucose, amino acids, nucleosides; P³²-labeled inorganic phosphate; S³⁵ labeled inorganic sulfate, sulfur amino acids.

2. Amount and Chemical Forms of Isotopes:

Maximum Amount on Hand Desired	*Chemical form	Byproduct Material (Element & Mass Number)
18 mCi	Inorganic + organic	H3
35 mCi	Inorganic + organic	C14
5 mCi	Inorganic + organic	P32
5 mCi	Inorganic + organic	P33
20 mCi	Inorganic + organic	S35
8 mCi	Inorganic + organic	I125

*See previous section for examples of inorganic and organic forms.

3. Proposed Date(s) for Studies:

Radioisotopes will be used on a routine basis (i.e., daily) in Dr. Smith research laboratory. Experiments with radioisotopes will begin upon approval of this request to use isotopes; and will continue as long as Dr. Smith is employed at the University of Florida. Dr. Smith's laboratory consist of three rooms (#G047, G058, G053) and a coldroom (between #G052 and G058) in McCarty Hall.

4. Protective Clothing:

For experiments with C¹⁴, H³, P³², P³³, S³⁵, the laboratory personnel will wear washable white Dacron/cotton laboratory coats. Plastic (or rubber gloves) will be worn. When experiments are performed with P³² and I¹²⁵, a protective lead apron will be worn which covers from the neck to the knees, and plastic (or rubber gloves) always will be worn. For experiments with P³² and I¹²⁵, the personnel will also work behind a lead-glass shield and lead bricks.

5. Experiments Performed Within and Outside of Hood:

Experiments which involve the use of isotopes which might be converted to volatile forms will be performed in a hood* in Room G058 McCarty Hall. Examples, of this type of research work include the use of [C¹⁴] bicarbonate in experiments with enzymes (e.g., ribulose-1, 5-bisphosphate carboxylase) and whole cells (e.g., photosynthetic incorporation of HCO₃) in which bicarbonate might be in equilibrium with atmospheric CO₂; experiments with [C¹⁴] a glucose and other sugars which are rapidly metabolized to CO₂. The handling of open vessels of I¹²⁵ as free iodine proteins (i.e., antibodies) always will be performed in a hood.

Experiments with non-volatile radiolabeled compounds or with compounds which will not be metabolized to volatile compounds will be performed outside of a hood. However, in this latter case the radioactive material will be performed on fiberglass trays with raised edges so that any spillage of material is contained on the tray.

Luxel/Film/TLD badges will be worn on the protective clothing during handling of strong Beta-emitters of Gamma-emitters.

6. Precautions to Prevent Spread of Contamination:

Whenever possible, experiments with radioisotopes will be performed on fiberglass trays with raised edges in Rooms G047, G052, G058. These trays will be identified with radiation signs. All radioactive glassware, pipettes, etc., will be maintained on these trays until washed in a sink designated for washing of contaminated glassware in Room #G058. After each experiment, the area will be initially monitored with a survey meter for possible contamination and then swabbed with a piece of Kimwipe tissue for measurement of radioactivity in a scintillation solution and counter. Any areas of contamination which are detected will be washed with tissues containing suitable detergent (e.g. Isoclean) used for radioactive decontamination. Any major spill would be dealt with as described in Chapter 2 section IX Emergency Procedures of the Radiation Control Guide. In addition to surveying radiation areas after each experiment, a "routine area" survey around work and isotope storage areas will be performed. Permanent records will be maintained of all specific and routine radiation survey.

Radioactive materials will be stored in a locked freezer and/or refrigerator. The radioactive materials will be so labeled and stored in a labeled box separate from non-radioactive materials. The fumehood in Dr. Jones' laboratory (Room #2047) will be used until installation of ductwork for fumehood in Dr. Smith's laboratory is completed in January 1995)

7. Radioactive Waste Disposal:

The research being performed in Dr. Smith's laboratory will generate both dry and liquid waste. Dry waste will be stored in properly marked containers lined with a heavy paper bag. Liquid waste will be stored in marked polyethylene bottles, with tight-fitting caps in a fume hood. After each experiment the type and amount of radioisotope, as waste, will be recorded. P³² and I¹²⁵ waste will be stored in a fume hood behind lead bricks. Each waste container or bottle will be labeled with Dr. Smith's name, and the total type and amount of radioisotope present.

8. Laboratory and Personnel Monitoring:

All persons working in an area designated for radioisotope handling shall wear monitoring devices. These devices shall be Whole body Luxel/film/TLD badges and ring badges, depending on the type of isotope being used. Personnel monitoring devices shall be changed every month or periodically, as determined by the RADIATION CONTROL OFFICER.

Areas where radioisotopes were used will be monitored at the end of each day by doing a s wipe test and counting in scintillation fluid in a liquid scintillation counter. When gamma emitters (P³², I¹²⁵) are used a survey meter (Nuclear Chicago, Model 2650M, approved and calibrated by RADIATION CONTROL) may also be used. AT the end of each week the entire laboratory and exit points will be monitored. All survey results will be kept in a permanent log.