Radioisotopes and Life Processes (Revised)Kisieleski, Walter E.
Science
Radioisotopes and Life Processes (Revised)
Kisieleski, Walter E.
Biology; Radioactive tracers
The condition of close contact between cells and emulsion is achieved by
the technique of dip-coating autoradiography. In this process the glass
slide on which the cells are carried is dipped into a melted
photographic emulsion (see Figure 15a), a thin film of which clings to
the slide. After it has been dried, the slide is placed in a lighttight
box and kept in a refrigerator for the desired period of exposure,
usually several days or weeks. During this period disintegrating
radioactive atoms within the cells continue to emit beta particles,
which, in turn, produce a latent image in the overlying emulsion. After
the exposure is complete, the slide is developed and fixed like a
photographic plate, and a stain is applied which penetrates the emulsion
so that the outlines of the cells and their internal structures can be
seen. The fixing process removes all silver bromide that has not been
ionized so that the emulsion is reduced to a thin, transparent film of
gelatin covering the stained cells and containing only the clusters of
silver grains that were struck by the beta particles.
[Illustration: Figure 16 _Radioautographs of tumor cells. Above, tumor
cells and blood cells. Below, magnification of tumor cells._]
When the finished autoradiograph is examined under the microscope, it
will look like the radioautographs of tumor cells in Figure 16. In the
upper micrograph the tumor cells are the larger ones and the smaller
ones are blood cells. The dense structures in the center of the tumor
cells are nuclei. The cells were exposed to tritium-labeled thymidine,
and those synthesizing DNA at the time of exposure took up the thymidine
and became radioactive. They can be identified by the black dots
overlying the nuclei; the dots are the aggregates of silver grains
struck by the beta particles.
Notice that only the nuclei contain radioactivity; the reason for this
is that radioactive thymidine is incorporated only into DNA localized in
the nuclei of cells. This picture identifies not only the cells that
were making DNA at the time the label was administered but also the
cells that were destined to divide in the immediate future, since cells
synthesize DNA in preparation for cell division.
If we want to compare two populations of cells to find out which is
proliferating (dividing) more actively, counting the fraction of cells
labeled will give the number of cells synthesizing DNA in preparation
for cell division. Of course, a rough approximation of the proliferating
activity can be obtained by simply counting the number of cells actually
dividing. But with tritiated thymidine we can obtain not only much more
accurate measurements but also considerable information that cannot be
obtained by simply counting the number of cells in mitosis. We shall
discuss the cell cycle later on, but for the moment we should emphasize
that much of our knowledge of the cell cycle stems from the use of
high-resolution autoradiography.
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