Ultraviolet-based photoacoustic microscopy (UV-PAM) has recently been demonstrated as a promising tool to overcome the time-consuming sample preparation procedure in traditional pathological analysis. In order to achieve high-speed UVPAM for clinical usage, we implemented UV-PAM with a single-axis galvo mirror scanner. With our UV laser operating at a repetition rate of 55 kHz, our system produced images ~5.5 times faster than the previously reported point-by-point raster scanning based UV-PAM, with a lateral resolution of ~1.0 μm. Histology-like images of a mouse brain slice were acquired by our system, showing its potential as an intraoperative imaging tool for surgical margin assessment.
Fourier ptychography (FP) is a promising computational imaging technique that overcomes the physical space-bandwidth product (SBP) limit of a conventional microscope by applying angular diversity illuminations. However, to date, the effective imaging numerical aperture (NA) achievable with a commercial LED board is still limited to the range of 0.3–0.7 with a 4×/0.1NA objective due to the constraint of planar geometry with weak illumination brightness and attenuated signal-to-noise ratio (SNR). Thus the highest achievable half-pitch resolution is usually constrained between 500–1000 nm, which cannot fulfill some needs of high-resolution biomedical imaging applications. Although it is possible to improve the resolution by using a higher magnification objective with larger NA instead of enlarging the illumination NA, the SBP is suppressed to some extent, making the FP technique less appealing, since the reduction of field-of-view (FOV) is much larger than the improvement of resolution in this FP platform. Herein, in this paper, we initially present a subwavelength resolution Fourier ptychography (SRFP) platform with a hemispherical digital condenser to provide high-angle programmable plane-wave illuminations of 0.95NA, attaining a 4×/0.1NA objective with the final effective imaging performance of 1.05NA at a half-pitch resolution of 244 nm with a wavelength of 465 nm across a wide FOV of 14.60 mm2 , corresponding to an SBP of 245 megapixels. Our work provides an essential step of FP towards high-NA imaging applications without scarfing the FOV, making it more practical and appealing.
Fourier ptychographic microscopy (FPM) is a recently developed computational imaging technique that achieves gigapixel images with both high resolution and large field-of-view. In the current FPM experimental setup, the dark-field images with high-angle illuminations are easily overwhelmed by stray lights and background noises due to the low signal-to-noise ratio, thus significantly degrading the achievable resolution of the FPM approach. We provide an overall and systematic data preprocessing scheme to enhance the FPM’s performance, which involves sampling analysis, underexposed/overexposed treatments, background noises suppression, and stray lights elimination. It is demonstrated experimentally with both US Air Force (USAF) 1951 resolution target and biological samples that the benefit of the noise removal by these methods far outweighs the defect of the accompanying signal loss, as part of the lost signals can be compensated by the improved consistencies among the captured raw images. In addition, the reported nonparametric scheme could be further cooperated with the existing state-of-the-art algorithms with a great flexibility, facilitating a stronger noise-robust capability of the FPM approach in various applications.
Fourier ptychographic microscopy (FPM) is a recently proposed computational imaging technique with both high-resolution and wide field of view. In current FPM imaging platforms, systematic error sources come from aberrations, light-emitting diode (LED) intensity fluctuation, parameter imperfections, and noise, all of which may severely corrupt the reconstruction results with similar artifacts. Therefore, it would be unlikely to distinguish the dominating error from these degraded reconstructions without any preknowledge. In addition, systematic error is generally a mixture of various error sources in the real situation, and it cannot be separated due to their mutual restriction and conversion. To this end, we report a system calibration procedure, termed SC-FPM, to calibrate the mixed systematic errors simultaneously from an overall perspective, based on the simulated annealing algorithm, the LED intensity correction method, the nonlinear regression process, and the adaptive step-size strategy, which involves the evaluation of an error metric at each iteration step, followed by the re-estimation of accurate parameters. The performance achieved both in simulations and experiments demonstrates that the proposed method outperforms other state-of-the-art algorithms. The reported system calibration scheme improves the robustness of FPM, relaxes the experiment conditions, and does not require any preknowledge, which makes the FPM more pragmatic.