Optogenetic manipulation of cell migration with high spatiotemporal resolution using lattice lightsheet microscopy

Lattice lightsheet microscopy (LLSM) featuring three-dimensional recording is improved to manipulate cellular behavior with subcellular resolution through optogenetic activation (optoLLSM). A position-controllable Bessel beam as a stimulation source is integrated into the LLSM to achieve spatiotemporal photoactivation by changing the spatial light modulator (SLM) patterns. Unlike the point-scanning in a confocal microscope, the lattice beams are capable of wide-field optical sectioning for optogenetic activation along the Bessel beam path.We show that the energy power required for optogenetic activations is lower than 1 nW (or 24 mWcm(-2)) for time-lapses of CRY2olig clustering proteins, and membrane ruffling can be induced at different locations within a cell with subcellular resolution through light-triggered recruitment of phosphoinositide 3-kinase. Moreover, with the epidermal growth factor receptor (EGFR) fused with CRY2olig, we are able to demonstrate guided cell migration using optogenetic stimulation for up to 6 h, where 463 imaging volumes are collected, without noticeable cellular damages. Using a Bessel beam as a simulation source allows the use of lattice lightsheet microscopy for spatiotemporal control of photoactivation, illustrated by the control of cellular migration behavior.

Automated summary

This study improves lattice lightsheet microscopy (LLSM) to manipulate cellular behavior with subcellular resolution through optogenetic activation (optoLLSM). By integrating a position-controllable Bessel beam as a stimulation source, spatiotemporal photoactivation can be achieved by changing spatial light modulator (SLM) patterns. The study demonstrates the ability to control membrane ruffling and cell migration behavior using light-triggered recruitment of phosphoinositide 3-kinase and guided cell migration with low energy power.