3D-IM&ACT – Remote focusing for volumetric imaging in combination with 3D patterning of the light

Sculpting light in vivo for uncaging or optogenetics relies on calibration performed in a cuvette but not in vivo. One solution consisting in combining volumetric imaging with the patterned illumination for stimulation is not trivial as the objective needs to remain steady in the axial direction to keep a stable 3D light pattern using holography. The objective of the 3D-IM&ACT project is to set up fast volumetric imaging in combination with 3D patterning for monitoring the effects of optical manipulation onto the activity of neuronal circuits in vivo. The system 3D-IM&ACT will achieve simultaneous multiphoton imaging at 4 volumes per second over 5 planes with a volume of > 0.5mm3 for single cell resolution photostimulation with sub-millisecond temporal control of custom activity patterns. The performances of the 3D-IM&ACT microscope have been already validated by a concept publication relying on imaging only (Rupprecht et al., 2016) and will be tested in vivo on 3 applications: the transparent zebrafish larva, as well as in mouse first in vitro, and in vivo in the cortex. The technology has been and will be further transferred to the industrial partner 3i in Paris.