SOLS excitation

In a single-objective light-sheet microscope, how should we configure the excitation?

Contents

Adjusting light-sheet angle

It is important to match the light-sheet angle to the image plane, and (to a lesser extent) the lightsheet direction in the field of view (FOV). This can be done by adjusting the position of the excitation in the back focal plane of objective 1, for example by conjugating a gimbal mirror (or similar) to an intermediate image plane on some part of the excitation path. If the gimbal mirror has 'tip/tilt' adjustment, then one adjuster be arranged to align the light-sheet tilt to the image plane,

SOLS_excitation_angle_to_image.png
(.odp sketch)

and the other adjuster to align the light-sheet direction to the field of view:

SOLS_excitation_angle_to_center.png
(.odp sketch)

Note: It may be worth automating one or both of these adjustments. For example the light-sheet 'angle to image' adjuster can be used to chase thermal drift and accommodate different samples that may affect this alignment. The 'angle to direction' adjuster can be used to scan the light-sheet direction, which helps with sample absorption and scattering for more uniform intensity across the FOV.