Optical coherence tomography (OCT) with microscopic resolution is suited for the investigation of dynamic processes on cellular level. Most existing setups for microscopic OCT (mOCT) are able to acquire A-Scans at 100 kHz, hence they are suitable for displaying only B-Scans in real-time. We present an mOCT setup with a new high-speed spectrometer, which is capable of acquiring up to 600.000 A-scans/s. Customized software allows to image mOCT volumes with micrometer resolution at video rate. Therefore, we are able to visualize functional processes on a cellular level in three dimensions over time. Here, we present 2D and 4D data of ex vivo human lung. Recording tens of high-resolution OCT volumes per seconds becomes possible faster than previous setup for OCT with microscopic resolution.
Optical coherence tomography (OCT) images scattering tissues with 5 to 15 μm resolution. This is usually not sufficient for a distinction of cellular and subcellular structures. Increasing axial and lateral resolution and compensation of artifacts caused by dispersion and aberrations is required to achieve cellular and subcellular resolution. This includes defocus which limit the usable depth of field at high lateral resolution. OCT gives access the phase of the scattered light and hence correction of dispersion and aberrations is possible by numerical algorithms. Here we present a unified dispersion/aberration correction which is based on a polynomial parameterization of the phase error and an optimization of the image quality using Shannon’s entropy. For validation, a supercontinuum light sources and a costume-made spectrometer with 400 nm bandwidth were combined with a high NA microscope objective in a setup for tissue and small animal imaging. Using this setup and computation corrections, volumetric imaging at 1.5 μm resolution is possible. Cellular and near cellular resolution is demonstrated in porcine cornea and the drosophila larva, when computational correction of dispersion and aberrations is used. Due to the excellent correction of the used microscope objective, defocus was the main contribution to the aberrations. In addition, higher aberrations caused by the sample itself were successfully corrected. Dispersion and aberrations are closely related artifacts in microscopic OCT imaging. Hence they can be corrected in the same way by optimization of the image quality. This way microscopic resolution is easily achieved in OCT imaging of static biological tissues.
A holographic method for high-speed, noncontact photoacoustic tomography is introduced and evaluated. Relative changes of the object’s topography, induced by the impact of thermoelastic pressure waves, were determined at nanometer sensitivity without physical contact. The object’s surface was illuminated with nanosecond laser pulses and imaged with a high-speed CMOS camera. From two interferograms measured before and after excitation of the acoustic wave, surface displacement was calculated and then used as the basis for a tomographic reconstruction of the initial pressure caused by optical absorption. The holographic detection scheme enables variable sampling rates of the photoacoustic signal of up to 50 MHz. The total acquisition times for complete volumes with 230 MVoxel is far below 1 s. Measurements of silicone and porcine skin tissue phantoms with embedded artificial absorbers, which served as a model for human subcutaneous vascular networks, were possible. Three-dimensional reconstructions of the absorbing structures show details with a diameter of 310 μm up to a depth of 2.5 mm. Theoretical limitations and the experimental sensitivity, as well as the potential for in vivo imaging depending on the detection repetition rate, are analyzed and discussed.