Laser Options

Use options on the Lasers tab of the Resolve3D Settings window to configure and align a laser for a photokinetics experiment.

Open the Laser settings options by clicking the Settings icon on the Resolve3D window and clicking the Lasers tab.

Field	Description
Module Status	Indicates whether the optional attached laser module is Off or On.
Laser	View which lasers are currently installed in the Laser Module Chassis.
Status	View the states of the Lasers. Status displays Off, Standby, Warming Up, or Ready for each laser. You cannot use a laser in an experiment until Status displays Ready.
Power (%)	The control voltage value that determines laser power. A typical value is 20%. The Power value is the percentage of maximum control voltage. Increasing control voltage increases laser power; however, control voltage is not linearly related to laser power.
Laser to test	Use the drop-down list in this field to select a specific laser on which to run PK tests.
Show spot alignment target	Activate this check box to include a target alignment reticle on the displayed test image as shown below.
Image The Spot	Selecting this button opens the laser shutter and acquires a single image of the laser spot. The event duration is set using the Exposure field in Resolve3D.
Continuous	Selecting this button opens the laser shutter and acquires continuous images of the laser spot. Select Stop Imaging to discontinue the imaging process.
Do Test Bleach	Perform a bleach test using the current Power and PK Duration settings.
% Bleach	The value entered in this field displays a before and after comparison of the intensities at the bleach spot: 100x (Signal After/Signal Before).
Background	The value in this field provides a measurement of the signal (intensity value) after the bleach event.
TIRF Illumination Settings	Selecting this button opens the TIRF Illumination Settings window from which the following actions are available: TIRF Beam Depth – allows you to select a laser to adjust its evanescent wave depth. Block – sets the TIRF depth adjuster to a position that prohibits illumination of the sample. Epifluorescence – sets the TIRF depth adjuster for epifluorescence (the highest value of the TIRF depth slider). Restore TIRF Depth – restores the TIRF depth adjuster to the last-used TIRF position. Back Aperture Focus for Current Lens – focuses the laser on the back aperture of the objective resulting in a collimated illumination laser and an even TIRF depth across the field of view. This value should be set once for each installed objective. To make sure this position is set correctly for imaging, verify the proper objective is selected from the Resolve3D drop-down menu. The Locked check box (activated by default) keeps the slider from being inadvertently moved. Laser Path Splitter – sets the percent of laser light to be directed to the TIRF illumination path versus the PK path. Use Beam Concentrator - enables the laser beam concentrator when check box is selected.