A woofer–tweeter adaptive optical structured illumination microscope(AOSIM) is presented. By combining a low-spatial-frequency large-stroke deformable mirror(woofer) with a high-spatial-frequency low-stroke deformabl...A woofer–tweeter adaptive optical structured illumination microscope(AOSIM) is presented. By combining a low-spatial-frequency large-stroke deformable mirror(woofer) with a high-spatial-frequency low-stroke deformable mirror(tweeter), we are able to remove both large-amplitude and high-order aberrations. In addition, using the structured illumination method, as compared to widefield microscopy, the AOSIM can accomplish highresolution imaging and possesses better sectioning capability. The AOSIM was tested by correcting a large aberration from a trial lens in the conjugate plane of the microscope objective aperture. The experimental results show that the AOSIM has a point spread function with an FWHM that is 140 nm wide(using a water immersion objective lens with NA=1.1) after correcting a large aberration(5.9 μm peak-to-valley wavefront error with 2.05 μm RMS aberration). After structured light illumination is applied, the results show that we are able to resolve two beads that are separated by 145 nm, 1.62× below the diffraction limit of 235 nm. Furthermore, we demonstrate the application of the AOSIM in the field of bioimaging. The sample under investigation was a green-fluorescentprotein-labeled Drosophila embryo. The aberrations from the refractive index mismatch between the microscope objective, the immersion fluid, the cover slip, and the sample itself are well corrected. Using AOSIM we were able to increase the SNR for our Drosophila embryo sample by 5×.展开更多
We discuss the implementation and performance of an adaptive optics(AO)system that uses two cascaded deformable phase plates(DPPs),which are transparent optofluidic phase modulators,mimicking the common woofer/tweeter...We discuss the implementation and performance of an adaptive optics(AO)system that uses two cascaded deformable phase plates(DPPs),which are transparent optofluidic phase modulators,mimicking the common woofer/tweeter-type astronomical AO systems.One of the DPPs has 25 electrodes forming a keystone pattern best suited for the correction of low-order and radially symmetric modes;the second device has 37 hexagonally packed electrodes better suited for high-order correction.We also present simulation results and experimental validation for a new open-loop control strategy enabling simultaneous control of both DPPs,which ensures optimum correction for both large-amplitude low-order,and complex combinations of low-and high-order aberrations.The resulting system can reproduce Zernike modes up to the sixth radial order with stroke and fidelity up to twice better than what is attainable with either of the DPPs individually.The performance of the new AO configuration is also verified in a custom-developed fluorescence microscope with sensorless aberration correction.展开更多
基金UC Office of the President(MR-15-327968)National Science Foundation(NSF)(1353461)National Institutes of Health(NIH)(R21MH101688)
文摘A woofer–tweeter adaptive optical structured illumination microscope(AOSIM) is presented. By combining a low-spatial-frequency large-stroke deformable mirror(woofer) with a high-spatial-frequency low-stroke deformable mirror(tweeter), we are able to remove both large-amplitude and high-order aberrations. In addition, using the structured illumination method, as compared to widefield microscopy, the AOSIM can accomplish highresolution imaging and possesses better sectioning capability. The AOSIM was tested by correcting a large aberration from a trial lens in the conjugate plane of the microscope objective aperture. The experimental results show that the AOSIM has a point spread function with an FWHM that is 140 nm wide(using a water immersion objective lens with NA=1.1) after correcting a large aberration(5.9 μm peak-to-valley wavefront error with 2.05 μm RMS aberration). After structured light illumination is applied, the results show that we are able to resolve two beads that are separated by 145 nm, 1.62× below the diffraction limit of 235 nm. Furthermore, we demonstrate the application of the AOSIM in the field of bioimaging. The sample under investigation was a green-fluorescentprotein-labeled Drosophila embryo. The aberrations from the refractive index mismatch between the microscope objective, the immersion fluid, the cover slip, and the sample itself are well corrected. Using AOSIM we were able to increase the SNR for our Drosophila embryo sample by 5×.
文摘We discuss the implementation and performance of an adaptive optics(AO)system that uses two cascaded deformable phase plates(DPPs),which are transparent optofluidic phase modulators,mimicking the common woofer/tweeter-type astronomical AO systems.One of the DPPs has 25 electrodes forming a keystone pattern best suited for the correction of low-order and radially symmetric modes;the second device has 37 hexagonally packed electrodes better suited for high-order correction.We also present simulation results and experimental validation for a new open-loop control strategy enabling simultaneous control of both DPPs,which ensures optimum correction for both large-amplitude low-order,and complex combinations of low-and high-order aberrations.The resulting system can reproduce Zernike modes up to the sixth radial order with stroke and fidelity up to twice better than what is attainable with either of the DPPs individually.The performance of the new AO configuration is also verified in a custom-developed fluorescence microscope with sensorless aberration correction.