All Research Areas

Imaging through complex media

The imaging performance of an optical microscope can be degraded by sample-induced aberrations. A general strategy to undo the effect of these aberrations is to apply wavefront correction with a deformable mirror (DM). In most cases the DM is placed conjugate to the microscope pupil, called pupil adaptive optics (AO). When the aberrations are spatially variant an alternative configuration involves placing the DM conjugate to the main source of aberrations, called conjugate AO. We have provided a theoretical and experimental comparison of both configurations for the simplified case where spatially variant aberrations are produced by a well-defined phase screen. Conjugate AO is found to provide a significant FOV advantage, which we xperimentally verified with standard widefield microscopy and with 2-photon microscopy.

We have also implemented a fast closed-loop feedback implementation of AO that requires no guide stars, where the sample itself serves as the reference. Several features of our implementation are new. First, it is based on a high-resolution, single-shot wavefront sensor that is compatible with extended samples. Second, it is applied to widefield (i.e., nonscanning) microscopy in a conjugate AO configuration that increases field of view. Third, it makes use of a fast algorithm to identify sample-induced aberrations using illumination from an arbitrarily shaped source. We present the principle of our technique and proof-of-concept experimental demonstrations.

Finally, we have characterized the spectral decorrelation that arises from the propagation of polychromatic light through complex media.


Publications Related to this Research Area

Widefield fluorescence microscopy with sensor-based conjugate adaptive optics using oblique back illumination

Jiang Li, Thomas G. Bifano, Jerome Mertz,

Journal of Biomedical Optics

We describe a wavefront sensor strategy for the implementation of adaptive optics (AO) in microscope applications involving thick, scattering media. The strategy is based on the exploitation of multiple scattering to provide oblique back illumination of the wavefront-sensor focal plane, enabling a simple and direct measurement of the flux-density tilt angles caused by aberrations at this plane. Advantages of the sensor are that it provides a large measurement field of view (FOV) while requiring no guide star, making it particularly adapted to a type of AO called conjugate AO, which provides a large correction FOV in cases when sample-induced aberrations arise from a single dominant plane (e.g., the sample surface). We apply conjugate AO here to widefield (i.e., nonscanning) fluorescence microscopy for the first time and demonstrate dynamic wavefront correction in a closed-loop implementation.

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Conjugate adaptive optics in widefield microscopy with an extended-source wavefront sensor

J. Li, D. R. Beaulieu, H. Paudel, R. Barankov, T. G. Bifano, and J. Mertz,

Optica

Subsurface microscopy is often limited by poor image quality due to sample-induced aberrations. Adaptive optics (AO) can counter such aberrations, though generally over limited fields of view. In most applications, AO is either slow or requires a “guide star” in the sample to serve as a localized reference target. We describe a fast closed-loop feedback implementation of AO that requires no guide stars, where the sample itself serves as the reference. Several features of our implementation are new. First, it is based on a high-resolution, single-shot wavefront sensor that is compatible with extended samples. Second, it is applied to widefield (i.e., nonscanning) microscopy in a conjugate AO configuration that increases field of view. Third, it makes use of a fast algorithm to identify sample-induced aberrations using illumination from an arbitrarily shaped source. We present the principle of our technique and proof-of-concept experimental demonstrations.

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Axial range of conjugate adaptive optics in two-photon microscopy

H. P. Paudel, J. Taranto, J. Mertz, and T. Bifano,

Optics Express

We describe an adaptive optics technique for two-photon microscopy in which the deformable mirror used for aberration compensation is positioned in a plane conjugate to the plane of the aberration. We demonstrate in a proof-of-principle experiment that this technique yields a large field of view advantage in comparison to standard pupil-conjugate adaptive optics. Further, we show that the extended field of view in conjugate AO is maintained over a relatively large axial translation of the deformable mirror with respect to the conjugate plane. We conclude with a discussion of limitations and prospects for the conjugate AO technique in two-photon biological microscopy.

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Field of view advantage of conjugate adaptive optics in microscopy applications

J. Mertz, H. Paudel, and T. G. Bifano,

Applied Optics

The imaging performance of an optical microscope can be degraded by sample-induced aberrations. A general strategy to undo the effect of these aberrations is to apply wavefront correction with a deformable mirror (DM). In most cases the DM is placed conjugate to the microscope pupil, called pupil adaptive optics (AO). When the aberrations are spatially variant an alternative configuration involves placing the DM conjugate to the main source of aberrations, called conjugate AO. We provide a theoretical and experimental comparison of both configurations for the simplified case where spatially variant aberrations are produced by a well-defined phase screen. We pay particular attention to the resulting correction field of view (FOV). Conjugate AO is found to provide a significant FOV advantage. While this result is well known in the astronomical community, our goal here is to recast it specifically for the optical microscopy community.

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Focusing polychromatic light through strongly scattering media

H. P. Paudel, C. Stockbridge, J. Mertz, T. Bifano,

Optics Express

We demonstrate feedback-optimized focusing of spatially coherent polychromatic light after transmission through strongly scattering media, and describe the relationship between optimized focus intensity and initial far-field speckle contrast. Optimization is performed using a MEMS spatial light modulator with camera-based or spectrometer-based feedback. We observe that the spectral bandwidth of the optimized focus depends on characteristics of the feedback signal. We interpret this dependence as a modification in the number of independent frequency components, or spectral correlations, transmitted by the sample, and introduce a simple model for polychromatic focus enhancement that is corroborated by experiment with calibrated samples.

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Conjugate versus pupil AO

Demonstration of the field of view advantage of conjugate (left) versus pupil (right) AO using 2-photon microscopy.

Closed-loop conjugate AO

Demonstration of real-time, closed-loop, conjugate AO using a PAW wavefront sensor. Sample is elastic mouse cartilage.