Chemistry Reference
In-Depth Information
Figure 4.1 Single molecule observations in
Ry
when bound to the channels. (b
e) Fluorescence
images of the same field. The upper panel (b, c)
shows images recorded by excitation with a red
laser (633 nm). The lower panel (d, e) show
excitation with both a green (532 nm) and red
(633 nm) laser. (b) and (d) show images of
Cy5
-
RyR2 binding. (a) Schematic representation
of the experiment. The receptor channels were
immobilized on the glass. Fluorescent ryanodine
molecules were not excited (
-
) when they were
distant from the glass surface. Excitation occurs
at a distance of less than 200 nm. However, they
cannot be visualized by video rate recording
because of their rapid three-dimensional thermal
motion. They were visualized as bright spots only
$
RyR2 while (c) and (e) represent
BodipyFL
-
RyR2 binding
duration measured in the presence of 10 nM
Ry and 10
Ry. (f) Histogram of Ry
-
-
MCa
2 รพ
.
m
the base of the chamber was coated with a thin layer of agarose. The bilayers
were positioned so as to make contact with the agarose layer. The TIRF microscope
allowed us to visualize single
fluorophores in the membrane by reducing
the background noise and also to detect single
fluorescent molecules in the
