Time-division beam multiplexing in Scanbox

Scanbox now allows interleaved scanning using tunable/fixed laser sources. If you are using such a laser (eg, Coherent Discovery TPC) the sliders in the Laser panel are configured to control the power of the tunable (left slider) and fixed (right slider) sources.

Either before or during scanning one can choose to alternate stimulation with the two sources by clicking the Mux checkbox in PMT Selection, Gain and Visualization panel. When active, the tunable laser source is used to scan the even frames and the fixed source is used to scan the odd frames. In this mode, one can toggle the frames being displayed by a toggle button located below the Mux checkbox. While one can visualize either odd or even frames during acquisition all the frames are being saved. Multiplexing works both in unidirectional and bidirectional modes.

This tool can be useful in two color imaging experiments that require different excitation wavelengths and where there may be substantial overlap between excitation or emission between the different fluorophores. Multiplexing minimizes cross-talk and between channels and provides the data required to estimate the mixing matrix if one decides to do experiments with simultaneous stimulation under the exact same conditions.

Below is a brief video showing testing of this feature using pollen grains:

and a a corn stem slide:

The Optotune & volumetric imaging panel currently allows modulating the tunable laser with depth, but there is already an entry prepared to allow the simultaneous modulation of the fixed laser. We are implementing this feature next, which will generalize the ability of users to do experiments where, for example, 920nm is used to image at shallow depths and 1040nm is used to image deeper as done here.