Chapter 2
STORM Microscopy Operation
14
7
Measure the gap between the boundary surface of the
cover glass and the position of the observation target
(Z-axis direction) using the method below.
This step is not required for 2D-STORM.
(1) Step
1:
When the right dialog is displayed, select the
imaging laser (647 nm). Set the focus to the
boundary between the cover glass and the
specimen by using the PFS offset controller. (Adjust
the power to about 50% to 100% using the slider.)
Set the focus to the boundary surface of the cover
glass.
When setting the focus, move the XY stage and view, as
boundary marks, the dye molecules that are
non-specifically adsorbed onto the boundary surface of
the cover glass to identify the boundary surface.
After setting the focus, click [OK]. The XY stage
automatically returns to its original position.
(2) Step
2:
When the dialog shown on the right is displayed, set
the focus to the target imaging position by using the
PFS offset controller.
After setting the focus, click [OK]. The measured
gap (Z-axis direction) is automatically saved and
used in analysis.
When acquisition is started, the dialog box shown on the
right is displayed. Do not perform any operation using this
dialog box until image acquisition has been completed.
Clicking [Events...], [Pause], or [Refocus] prevents the
acquisition of appropriate images for STORM analysis.
To halt the acquisition, click [Finish] or [Abort]. (If [Finish] is
clicked, the acquired images are saved. Be careful
because, if [Abort] is clicked, the acquired images will be
discarded.)
7-(1)
7-(2)
Cover glass
Boundary surface of
cover glass
Cover glass
Imaging target surface
