v.闪烁countingofb-emissions3 h，14，35，和（ v. scintillation countingofb-emissions3 h,14 c,35 s,and）.docVIP

Biochemistry 3723 Lecture 4b: Radioisotopes August 28, 2002 V. Scintillation Counting of ?- emissions: 3H, 14C, 35S, and 32P A. Process of scintillation: use 3H as an example: 1. ?- decays with energy between 0 and 0.0186 MeV (= 18.6 KeV) 2. If radioactive solute dissolved in appropriate solvent, collision of ?- with solvent transfers some of energy to solvent, “exciting” solvent molecule. a. non-bonding π electrons excited to higher state without loss of energy b. ?- continues to excite solvent until all energy of particle dissipated. c. ?-**** + S ? ?-*** + S*; ?-*** + S ? ?-** + S* ; etc.(* = energy) 3. Excited solvent transfers energy to fluor, a compound that accepts energy easily from S and decays to ground state with emission of light S* + F ? S + F*; F*?? F + light (h?) 4. Look at elements of overall process a. ?- emission ? photons takes 5 nsec. i. Yield ≈ 10 photons /KeV. Therefore most energetic 3H decay ≈ 186 photons ii. # photons ≈ linear w/energy of ?-. Average 5.5 KeV ≈ 55 photons b. Solvent--various aromatic compounds-- toluene, xylene, alkylbenzenes, alkylnapthalenes c. Fluor--various multicyclic compounds (aromatic) -delocalized electrons d. Scintillation Cocktail: Fluor dissolved in Solvent e. Additions to allow solution of aqueous samples-i.e. DMSO f. Many proprietary formulations (companies wont tell exactly how they are formulated, because then you wouldnt buy their expensive products.) B. The Scintillation Counter: Necessary parts—in your book (p 177) 1. Sample chamber--where you put the scintillation vial containing sample dissolved in scintillation coctail 2. Photomultiplier tubes a. Two of them see light flashes. b. Resolution time ≈ 20 nsec. Means absolute limit ≈ 5 x 107 cps 3. Pulse summation (counts # events) 4. Coincidence circuit a. Light flash must be seen by both PMTs at same time to be counted. i Theoretical minimum to be seen: 2 photons