2.7 X-LIGHT alignment
Focusing the pinholes
The spinning disk is in a plane conjugate to the image plane. As it spins, it scans the microscopic fluorescent
image point by point in real-time. It is essential that the CCD/sCMOS cameras be precisely focused on the
image plane (the pinholes) of the spinning disk in order to obtain a sharp image.
Turn on the bright field lamp of the microscope and send the light to the side port of the microscope. You
don’t need a sample on the microscope. Using your imaging software do the following:
(1) Select the GFP filter set in the light path
(2) Move spinning disk into light path (confocal mode)
(3) Stop the disk from spinning
(4) Switch the camera into live mode using the imaging software
(5) Adjust the exposure time to 50 ms and then adjust the bright field lamp power until a non-saturated
image of the pinholes can be observed
(6) The pinholes should be close to focus. The idea is to adjust the X-Light V2
TP
focusing lens (see
picture) until the pinholes are in focus and the image is sharp.
Remove the cover Rotate the hex key to focus
To Next Page