Method for combined /sup 3/H and /sup 14/C autoradiography with a single emulsion tested in cultured mammalian cells

Journal Article · Sat Jan 01 04:00:00 EST 1977 · Exp. Cell Res.; (United States)

DOI:https://doi.org/10.1016/0014-4827(77)90384-6· OSTI ID:5545708

Perdue, S W; Kimball, R F; Hsie, A W

A single-gelatin expanded film method for double-isotope autoradiography is described. A preliminary classification based upon silver-grain distribution is used to assign labeled cells to /sup 3/H only or with /sup 14/C classes. Optical sectioning combined with grain counting is employed to obtain ratios for classifying cells labeled with /sup 14/C into /sup 14/C only and /sup 3/H + /sup 14/C classes. The method has been tested with CHO-K1 cells in plateau-phase cultures using two 24-h labeling periods. The experimental design allowed for independent estimation of the expected frequencies of label classes under conditions that provided a wide range of possible label levels and combinations. Previous methods have used time-consuming applications of two emulsion layers and exposures to distinguish between cells labeled with /sup 3/H only or with /sup 14/C and do not identify the /sup 3/H + /sup 14/C class. A single-gelatin expanded film requires only one exposure and permits all label classes to be determined by an objective grain-counting procedure.

• Research Organization: Oak Ridge National Lab., TN
• OSTI ID: 5545708
• Journal Information: Exp. Cell Res.; (United States), Journal Name: Exp. Cell Res.; (United States) Vol. 107; ISSN ECREA
• Country of Publication: United States
• Language: English

Related Subjects

550301* -- Cytology-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ANIMALS
AUTORADIOGRAPHY
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
CARBON 14
CARBON ISOTOPES
CELL CULTURES
COLLOIDS
DISPERSIONS
EMULSIONS
EVEN-EVEN NUCLEI
HISTOLOGICAL TECHNIQUES
HYDROGEN ISOTOPES
ISOTOPES
LIGHT NUCLEI
MAMMALS
NUCLEI
ODD-EVEN NUCLEI
RADIOISOTOPES
TRITIUM
VERTEBRATES
YEARS LIVING RADIOISOTOPES