Noninvasive functional imaging of molecular and cellular processes of vision may have immense impact on research and clinical diagnostics. Although suitable intrinsic optical signals (IOSs) have been observed ex vivo and in immobilized animals in vivo, detecting IOSs of photoreceptor activity in living humans was cumbersome and time consuming. Here, we observed clear spatially and temporally resolved changes in the optical path length of the photoreceptor outer segment as a response to an optical stimulus in the living human eye. To witness these changes, we evaluated phase data obtained with a parallelized and computationally aberration-corrected optical coherence tomography system. The noninvasive detection of optical path length changes shows neuronal photoreceptor activity of single cones in living human retina, and therefore, it may provide diagnostic options in ophthalmology and neurology and could provide insights into visual phototransduction in humans.functional optical coherence tomography | intrinsic optical signals | imaging | adaptive optics | phototransduction

Certain topics in research and advancements in medical diagnostics may benefit from improved temporal and spatial resolution during non-invasive optical imaging of living tissue. However, so far no imaging technique can generate entirely diffraction-limited tomographic volumes with a single data acquisition, if the target moves or changes rapidly, such as the human retina. Additionally, the presence of aberrations may represent further difficulties. We show that a simple interferometric setup–based on parallelized optical coherence tomography–acquires volumetric data with 10 billion voxels per second, exceeding previous imaging speeds by an order of magnitude. This allows us to computationally obtain and correct defocus and aberrations resulting in entirely diffraction-limited volumes. As demonstration, we imaged living human retina with clearly visible nerve fiber layer, small capillary networks, and photoreceptor cells. Furthermore, the technique can also obtain phase-sensitive volumes of other scattering structures at unprecedented acquisition speeds.

We demonstrate a new noninvasive method to assess biomechanical properties of the retinal vascular system. Phase-sensitive full-field swept-source optical coherence tomography (PhS-FF-SS-OCT) is used to investigate retinal vascular dynamics at unprecedented temporal resolution. The motion of retinal tissue that is induced by expansion of the vessels therein is measured with an accuracy of about 10 nm. The pulse shapes of arterial and venous pulsations, their temporal delays, as well as the frequency-dependent pulse propagation through the capillary bed, are determined. For the first time, imaging speed and motion sensitivity are sufficient for a direct measurement of pulse waves propagating with more than 600 mm/s in retinal vessels of a healthy young subject.

Swept-source optical coherence tomography (SS-OCT) is sensitive to sample motion during the wavelength sweep, which leads to image blurring and image artifacts. In line-field and full-field SS-OCT parallelization is achieved by using a line or area detector, respectively. Thus, approximately 1000 lines or images at different wavenumbers are acquired. The sweep duration is identically with the acquisition time of a complete B-scan or volume, rendering parallel SS-OCT more sensitive to motion artifacts than scanning OCT. The effect of axial motion on the measured spectra is similar to the effect of non-balanced group velocity dispersion (GVD) in the interferometer arms. It causes the apparent optical path lengths in the sample arm to vary with the wavenumber. Here we propose the cross-correlation of sub-bandwidth reconstructions (CCSBR) as a new algorithm that is capable of detecting and correcting the artifacts induced by axial motion in line-field or full-field SS-OCT as well as GVD mismatch in any Fourier-domain OCT (FD-OCT) setup. By cross-correlating images which were reconstructed from a limited spectral range of the interference signal, a phase error is determined which is used to correct the spectral modulation prior to the calculation of the A-scans. Performance of the algorithm is demonstrated on in vivo full-field SS-OCT images of skin and scanning FD-OCT of skin and retina.

Holoscopy is a tomographic imaging technique that combines digital holography and Fourier-domain optical coherence tomography (OCT) to gain tomograms with diffraction limited resolution and uniform sensitivity over several Rayleigh lengths. The lateral image information is calculated from the spatial interference pattern formed by light scattered from the sample and a reference beam. The depth information is obtained from the spectral dependence of the recorded digital holograms. Numerous digital holograms are acquired at different wavelengths and then reconstructed for a common plane in the sample. Afterwards standard Fourier-domain OCT signal processing achieves depth discrimination. Here we describe and demonstrate an optimized data reconstruction algorithm for holoscopy which is related to the inverse scattering reconstruction of wavelength-scanned full-field optical coherence tomography data. Instead of calculating a regularized pseudoinverse of the forward operator, the recorded optical fields are propagated back into the sample volume. In one processing step the high frequency components of the scattering potential are reconstructed on a non-equidistant grid in three-dimensional spatial frequency space. A Fourier transform yields an OCT equivalent image of the object structure. In contrast to the original holoscopy reconstruction with backpropagation and Fourier transform with respect to the wavenumber, the required processing time does neither depend on the confocal parameter nor on the depth of the volume. For an imaging NA of 0.14, the processing time was decreased by a factor of 15, at higher NA the gain in reconstruction speed may reach two orders of magnitude.