Cell Culture Dishes for Microbeam Irradiation
cell culture dish.
Cell culture irradiations are the primary form
of microbeam irradiations that are performed at RARAF. The cells
are plated on a modified culture dish where a 1/4" hole has been
drilled in the center to act as a culture well. The bottom of
the well is covered with 3 micron polypropylene film where the
cells are plated. This thin polypropylene allows the chosen radiation
to get through to the cells while allowing them to be placed upright
on the microbeam end stations with minimal distance between the
dish bottom and beam exit window.
We have drafted instruction sheets
on preparing microbeam
dishes and plating
cells for microbeam irradiation.
A list of cell lines that have been previously used for microbeam irradiation is available
The dish making and handling procedures have
been demonstrated in a video for our microbeam training course.
Video for Dish Handling
with multiple parts labeled with separate fluorophores.
DNA (blue), mitochondria (red) and Actin fibers (green)
are shown here.
The dishes provide a stable base on
which the cells can be plated and maintained throughout the microbeam
irradiations. The cells are primarily imaged through fluorescent
staining with Hoechst 33342 DNA binding vital stain to target
nuclei to be hit or not hit with the irradiation. Labeling of
multiple parts of cells for irradiation and/or observation is
also available. A picture of multi-color stained cells in culture
is shown here. Other imaging modalities are available if desired
and are discussed on the Microbeam
Online Imaging page.