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Extended depth-of-field microscopy with a high-speed deformable mirror

Wide-field fluorescence microscopy is generally limited to either small volumes or low temporal resolution. We present a microscope add-on that provides fast, light-efficient extended depth-of-field (EDOF) using a deformable mirror of update rate 20kHz. Out-of-focus contributions in the raw EDOF images are suppressed with a deconvolution algorithm derived directly from the microscope 3D optical transfer function. Demonstrations of the benefits of EDOF microscopy are shown with GCaMP-labeled mouse brain tissue.