Journal
APPLIED PHYSICS LETTERS
Volume 118, Issue 2, Pages -Publisher
AMER INST PHYSICS
DOI: 10.1063/5.0035183
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- Army Research Office (ARO) [W911NF-16-1-0014]
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By using nonlinear optical techniques to control bidirectional optogenetic switches, the selectivity of the photoactivation step can be enhanced beyond the effects of traditional linear excitation with monochromatic CW light. Simulating spectral features and employing stimulated depletion quenching as a nonlinear optical strategy can effectively improve the dynamic range of bidirectional switches.
Conventionally, bidirectional optogenetic switches are controlled with linear excitation (e.g., monochromatic CW light), which has serious limitations when there is a spectral overlap between the on and off states of the switch. The spectral crosstalk lowers the selectivity of the photoactivation step, resulting in a moderate photoequilibrium value and a poor dynamic range for the switch. Using stimulated depletion quenching (SDQ), which is a nonlinear optical strategy similar to STED, we demonstrate enhanced photoactivation selectivity on one side of the switch, thus shifting the photoequilibrium beyond what is achievable with CW light. The discussion is built around Cph8 as a prototypical bidirectional optogenetic switch lacking complete photoreversibility upon CW excitation because of the spectral crosstalk. We use two fluorescent dyes as analogues to emulate the key spectral feature of the on and off states of Cph8; in this way, we focus on the initial photoactivation step and abstract from the complicated subsequent dynamics. By applying SDQ to a mixture of the dyes, we enhance the selectivity of the photoactivation beyond the linear regime. Increased selectivity of the photoactivation step via nonlinear optical techniques should translate into an improved dynamic range for a broad variety of bidirectional switches. These experiments provide a basis to further expand the foundations for non-conventional optical control methods of biological switching.
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