抄録
A novel method was developed to estimate the
relative amounts of total and local attachmin
in immunostained mouse peritoneal macrophages
using confocal laser microscopy. Macrophages were
made to adhere to an uncoated glass surface;
attachmin in the cells was immunostained with a
polyclonal anti-attachmin antibody and a FITC-labeled
secondary antibody after treatment with or
without Nonidet P-40. Fluorescence in thin optical
sections through the vertical and tangential levels of
the glass surface was quantitatively measured with
a confocal laser microscope; relative amounts of
total and local attachmin per cell were estimated
during the cell adhesion to glass. Using this method
revealed that the attachmin was distributed all over
the macrophage plasma membranes homogeneously;
at the free surface of the cell, approximately
half of the ≥ 300-kDa attachmin polymer was found
to protrude outside the cell through the plasma
membranes. This simple method will be used for
quantitative analysis of the distribution and localization
of various insoluble antigenic components
within cells.
