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This chapter is intended to provide a brief overview of the optics of surgical microscopes and rigid endoscopes, with the aim of providing the reader with the principles dictating the nature of surgical visualization when either of the visual control systems is used. It is not by any means geared toward elaborating on the detailed optical physics of these systems, which is beyond the scope and objective of this chapter., (© 2024. The Author(s), under exclusive license to Springer Nature Switzerland AG.)

The maker movement has reached the optics labs, empowering researchers to create and modify microscope designs and imaging accessories. 3D printing has a disruptive impact on the field, improving accessibility to fabrication technologies in additive manufacturing. This approach is particularly useful for rapid, low-cost prototyping, allowing unprecedented levels of productivity and accessibility. From inexpensive microscopes for education such as the FlyPi to the highly complex robotic microscope OpenFlexure, 3D printing is paving the way for the democratization of technology, promoting collaborative environments between researchers, as 3D designs are easily shared. This holds the unique possibility of extending the open-access concept from knowledge to technology, allowing researchers everywhere to use and extend model structures. Here, it is presented a review of additive manufacturing applications in optical microscopy for life sciences, guiding the user through this new and exciting technology and providing a starting point to anyone willing to employ this versatile and powerful new tool., (© 2021 The Authors. Advanced Biology published by Wiley-VCH GmbH.)

The rise of microscopy in the seventeenth century allowed scientists to discover a new world of microorganisms and achieve great physiological advances. One of the first microscopes of the epoch was Antonie van Leeuwenhoek's microscope, a deceptively simple device that contains a single ball lens housed in a metal plate allowing the observation of samples at up to ×250 magnification. Such magnification was much greater than that achieved by rudimentary compound microscopes of the era, allowing for the discovery of microscopic, single-celled life, an achievement that marked the study of biology up to the nineteenth century. Since Leeuwenhoek's design uses a single ball lens, it is possible to fabricate variations for educational activities in physics and biology university and high school classrooms. A fundamental problem, however, with home-built microscopes is that it is difficult to work with glass. We developed a simple protocol to make ball lenses of glass and gelatin with high magnification that can be done in a university/high school classroom, and we designed an optimized support for focusing and taking photographs with a smartphone. The protocol details a simple, easily accessible, low-cost, and effective tool for the observation of microscopic samples, possible to perform anywhere without the need for a laboratory or complex tools. Our protocol has been implemented in classrooms in Chile to a favorable reception.

We introduce a random-access parallel (RAP) imaging modality that uses a novel design inspired by a Newtonian telescope to image multiple spatially separated samples without moving parts or robotics. This scheme enables near-simultaneous image capture of multiple petri dishes and random-access imaging with sub-millisecond switching times at the full resolution of the camera. This enables the RAP system to capture long-duration records from different samples in parallel, which is not possible using conventional automated microscopes. The system is demonstrated by continuously imaging multiple cardiac monolayer and Caenorhabditis elegans preparations., Competing Interests: MA, SM, BS, AT, KR, DB, ST, AC, GB No competing interests declared, (© 2021, Ashraf et al.)

Significance: Surgical microscopes provide adjustable magnification, bright illumination, and clear visualization of the surgical field and have been increasingly used in operating rooms. State-of-the-art surgical microscopes are integrated with various imaging modalities, such as optical coherence tomography (OCT), fluorescence imaging, and augmented reality (AR) for image-guided surgery., Aim: This comprehensive review is based on the literature of over 500 papers that cover the technology development and applications of surgical microscopy over the past century. The aim of this review is threefold: (i) providing a comprehensive technical overview of surgical microscopes, (ii) providing critical references for microscope selection and system development, and (iii) providing an overview of various medical applications., Approach: More than 500 references were collected and reviewed. A timeline of important milestones during the evolution of surgical microscope is provided in this study. An in-depth technical overview of the optical system, mechanical system, illumination, visualization, and integration with advanced imaging modalities is provided. Various medical applications of surgical microscopes in neurosurgery and spine surgery, ophthalmic surgery, ear-nose-throat (ENT) surgery, endodontics, and plastic and reconstructive surgery are described., Results: Surgical microscopy has been significantly advanced in the technical aspects of high-end optics, bright and shadow-free illumination, stable and flexible mechanical design, and versatile visualization. New imaging modalities, such as hyperspectral imaging, OCT, fluorescence imaging, photoacoustic microscopy, and laser speckle contrast imaging, are being integrated with surgical microscopes. Advanced visualization and AR are being added to surgical microscopes as new features that are changing clinical practices in the operating room., Conclusions: The combination of new imaging technologies and surgical microscopy will enable surgeons to perform challenging procedures and improve surgical outcomes. With advanced visualization and improved ergonomics, the surgical microscope has become a powerful tool in neurosurgery, spinal, ENT, ophthalmic, plastic and reconstructive surgeries.

Background: Although recent developments in methodologies for light microscopy have enabled imaging of fine biological structures, such imaging is often accompanied by two types of problems. One is a tilting of the specimen with respect to the x-y plane (i.e. rotation around the x- or y-axis) such that the sample is not perpendicular to the optical z-axis, and the other is rotation around the z-axis that precludes optimal orientations for imaging and experimentation. These rotation problems can cause optical aberrations and hamper imaging experiments, even when the angular difference from the ideal position is small., New Method: In order to correct for these practical issues, we have developed a specimen holder with 3-axis (x-y-z) rotation for an inverted light microscope. This allows for full-range rotations of 2-4° for x-, y-axes, ~24° for z-axis, and a small-angle control of <0.1° for either axis., Results: Using this device, we observed the cultured hippocampal neurons stained by immunofluorescence for a dendritic marker, or the sub-resolution fluorescent beads plated on a glass coverslip. The rotations and associated problems could be manipulated, while viewing the specimens by laser-scanning confocal microscopy., Comparison With Existing Methods: This tilting/rotation device is easily manufactured and installed on a conventional microscope stage without requiring changes to the existing optical components. Similar devices with full capability have not been available., Conclusions: It will be useful for imaging experiments with biomedical applications., (Copyright © 2013 Elsevier B.V. All rights reserved.)

A new method of polarized light analysis is described in which a highly sensitive electronic detector specific for birefringence is used to identify the crystalline axes of an object and then measure its phase retardation due to birefringence. The microscopic system employed in the method consists of an electronic birefringence detection system (BDS), a microscope with strain-free lenses, and a driven stage for passing the specimen at appropriate velocities across the image of an aperture placed at the field stop and imaged in the specimen plane by the condenser. The detector registers retardations directly as voltage at a constant deflection sensitivity of ca. 1.1 v per angstrom unit over a range of 120 angstrom units. The basal rms noise level is 0.002 A for a spot 36 micro in diameter formed by a 95 x, N. A. 1.25 objective pair, and increases in proportion to the reciprocal of the diameter of the scanning spot. The increase in noise with high resolution scanning can be offset by increasing the instrumental time constant, which is adjustable in decades between 0.004 and 0.4 seconds. A number of difficult problems in high extinction polarization microscopy are avoided by the use of modulated light and a rapid electronic detector. For example: (a) The measured distribution of birefringence is unaffected by the usual diffraction anomaly; therefore polarization rectifiers are not required. (b) The detector is selective for birefringence, so that there is no problem in separating contrast due to different optical properties (e.g. dichroism, light scattering). (c) The speed and sensitivity are both increased by between one and two orders of magnitude over that attainable by visual or photographic methods, thereby rendering a vast number of weakly birefringent, light-scattering, and motile objects readily analyzable for the first time with polarized light.

This paper presents a microscopy technique that can perform snapshot depth resolved optical imaging in the same manner as A-scan in ultrasound imaging and optical coherence tomography. In this technique, a laser line along the axial dimension is used to illuminate a sample to create a fluorescent line object. By transforming the line object along the axial dimension (Z) to a ring image on the lateral dimensions (X-Y) using a full cone mirror, common optics can be used to relay and acquire the ring image precisely. Then, by converting half of the ring image back to a line image using a half cone mirror, the opening side of the half cone mirror allows the line image, which contains the full depth resolved information of the line object, to be taken in one snapshot. This eliminates the requirement of axial scanning in traditional depth resolved imaging techniques such as confocal microscopy to obtain the same information. The technique is demonstrated by imaging fluorescent microspheres of different diameters. This technique offers a simple alternative to traditional depth resolved imaging techniques such as confocal microscopy and light sheet microscopy. It is particularly useful in imaging samples with multiple layers in which multiple A-scans or a few B-scans are sufficient to represent the entire sample. [ABSTRACT FROM AUTHOR]

Wide-field mid-infrared (MIR) hyperspectral imaging offers a promising approach for studying heterogeneous chemical systems due to its ability to independently characterize the molecular properties of different regions of a sample. However, applications of wide-field MIR microscopy are limited to spatial resolutions no better than ∼1 μm. While methods exist to overcome the classical diffraction limit of ∼λ/2, chromatic aberration from transmissive imaging reduces the achievable resolution. Here we describe the design and implementation of a simple MIR achromatic lens combination that we believe will aid in the development of resolution-enhanced wide-field MIR hyperspectral optical and chemical absorption imaging. We also examine the use of this doublet lens to image through polystyrene microspheres, an emerging and simple means for enhancing spatial resolution. [ABSTRACT FROM AUTHOR]

Optical superresolution microscopy is an important field, where nonlinear optical processes or prior information is used to defeat the classical diffraction limit of light. Quantum correlation microscopy uses photon arrival statistics from single photon emitters to aid in the determination of properties including the number of emitters and their relative brightness. Here we model quantum correlation microscopy in the few emitter regime, i.e. around four single photon emitters below the diffraction limit. We use the Akaike Information Criterion to determine the number of emitters and we vary the relative contributions of intensity to quantum correlation information to determine contribution that provides optimal imaging. Our results show diffraction unlimited performance and a change in localisation scaling behaviour dependent on emitter closeness. [ABSTRACT FROM AUTHOR]

We propose a compact, portable, and low-cost holographic microscope designed for the characterization of micrometric particles suspended in a liquid. This system is built around a commercial optical microscope by substituting its illumination source (a light-emitting diode) with a collimated laser beam. Similarly, a quartz flow cell replaces the microscope glass slide using a 3D-printed custom mount. With the hardware presented in this paper, the holographic imaging of the electromagnetic fields emitted by the particles that intercept the laser beam achieves a resolution close to that of optical microscopes but with a greater depth of field. Several morphological and optical features can be extracted from the holograms, including particle projected section, aspect ratio, and extinction cross-section. Additionally, we introduce a remote system control that enables users to process the acquired holograms on a remote computational device. This work provides a comprehensive description of the methodology of image processing in holographic microscopy and a series of validation measurements conducted using calibrated particles. This technique is suitable for the characterization of airborne particles found in snow, firn, and ice; here we report experimental results obtained from Alpine ice cores.

SignificanceTraditional pathology workflow suffers from limitations including biopsy invasiveness, small fraction of large tissue samples being analyzed, and complex and time-consuming processing.AimWe address limitations of conventional pathology workflow through development of a laser microbiopsy device for minimally invasive harvest of sub-microliter tissue volumes. Laser microbiopsy combined with rapid diagnostic methods, such as virtual hematoxylin and eosin (H&E) imaging has potential to provide rapid minimally invasive tissue diagnosis.ApproachLaser microbiopsies were harvested using an annular shaped Ho:YAG laser beam focused onto the tissue surface. As the annulus was ablated, the tissue section in the center of the annulus was ejected and collected directly onto a glass slide for analysis. Cryogen spray cooling was used before and after laser harvest to limit thermal damage. Microbiopsies were collected from porcine skin and kidney. Harvested microbiopsies were imaged with confocal microscopy and digitally false colored to provide virtual H&E images.ResultsMicrobiopsies were successfully harvested from porcine skin and kidney. Computational and experimental results show the benefit of cryogen pre- and post-cooling to limit thermal damage. Virtual H&E images of microbiopsies retained observable cellular features including cell nuclei.ConclusionsLaser microbiopsy with virtual H&E imaging shows promise as a potential rapid and minimally invasive tool for biopsy and diagnosis.

Metasurfaces have attracted considerable attention because of their unique optical capabilities to control the fundamental properties of light, such as amplitude, phase, and polarization. The flat nature of metasurfaces can help reduce the complexities and bulk of conventional optical systems. After a decade of rapid progress, metasurfaces are close to maturity and have found their role in various optical applications. This review emphasizes the significant advancements and emerging applications of metasurfaces in biomedical optics, particularly focusing on beam shaping for laser treatments, light-sheet fluorescence microscopy, HiLo microscopy, and optical trapping. Looking forward, we discuss foreseeable challenges for integrating metasurfaces into biomedical, preclinical, and clinical systems. [ABSTRACT FROM AUTHOR]

SignificanceFollicular thyroid carcinoma carries a substantially poor prognosis due to its unique biological behavior and less favorable outcomes. In particular, fine-needle aspiration (FNA) biopsies, which play a key role in screening thyroid nodules, cannot differentiate benign from malignant follicular neoplasm.AimWe report on the use of hyperspectral Raman microscopy in combination with chemometric analysis for identifying and classifying single cells obtained from clinical samples of human follicular thyroid neoplasms.ApproachWe used a method intended to simulate the FNA procedure to obtain single cells from thyroid nodules. A total of 392 hyperspectral Raman images of single cells from follicular thyroid neoplasms were collected.ResultsMalignant cells were identified based on their intrinsic Raman spectral signatures with an overall diagnostic accuracy of up to 83.7%.ConclusionsOur findings indicate that hyperspectral Raman microscopy can potentially be developed into an ancillary test for analyzing single cells from thyroid FNA biopsies to better stratify "indeterminate" nodules and other cytologically challenging cases.

The objective of this research includes integration of high-resolution imaging through scattering medium, such as blood, into a disposable micro-endoscope. A fiber laser integrated into the micro-endoscope as part of its illumination channel, allows to project a tunable array of spots of light onto an object, that is located behind the scattering medium. We have a laser fiber as part of the illumination channel of a disposable micro-endoscope. By using proper optics, we convert the temporal modulation of the laser into spatial distribution. Thus, the result is generation of spatial spots when using a pulsed laser. The detection channel is a holographic recording of the collected back scattered light, that allows extraction of the electrical field. By time integrating the field we obtain the realization of the spatial array of illumination spots formed on top of the inspected object and behind the scattering medium. By changing the temporal modulation of the illumination laser (changing its temporal photonic signals), we can tune the positions of the spots in the illumination array. If the distance between the projected spots is larger than the imaging resolution, then by applying localization microscopy algorithms together with scanning of the position of the spots in the array, will yield a high-resolution reconstruction of the inspected object. We theoretically and experimentally demonstrate the discussed operation principle and show the potential of the proposed concept as a modality in medical endoscopic procedures. [ABSTRACT FROM AUTHOR]

Conventional microscopy systems have limited depth of field, which often necessitates depth scanning techniques hindered by light scattering. Various techniques have been developed to address this challenge, but they have limited extended depth of field (EDOF) capabilities. To overcome this challenge, this study proposes an end-to-end optimization framework for building a computational EDOF microscope that combines a 4f microscopy optical setup incorporating learned optics at the Fourier plane and a post-processing deblurring neural network. Utilizing the end-to-end differentiable model, we present a systematic design methodology for computational EDOF microscopy based on the specific visualization requirements of the sample under examination. In particular, we demonstrate that the metasurface optics provides key advantages for extreme EDOF imaging conditions, where the extended DOF range is well beyond what is demonstrated in state of the art, achieving superior EDOF performance. [ABSTRACT FROM AUTHOR]

Coherent Raman scattering (CRS) microscopy is an optical imaging technique with capabilities that could benefit a broad range of biomedical research studies. We reflect on the birth, rapid rise, and inescapable growing pains of the technique and look back on nearly four decades of developments to examine where the CRS imaging approach might be headed in the next decade to come. We provide a brief historical account of CRS microscopy, followed by a discussion of the challenges to disseminate the technique to a larger audience. We then highlight recent progress in expanding the capabilities of the CRS microscope and assess its current appeal as a practical imaging tool. New developments in Raman tagging have improved the specificity and sensitivity of the CRS technique. In addition, technical advances have led to CRS microscopes that can capture hyperspectral data cubes at practical acquisition times. These improvements have broadened the application space of the technique. The technical performance of the CRS microscope has improved dramatically since its inception, but these advances have not yet translated into a substantial user base beyond a strong core of enthusiasts. Nonetheless, new developments are poised to move the unique capabilities of the technique into the hands of more users.

Polarizing optical microscopy and electro optics are used to study a compound with constituent non-chiral molecules, DIO, known to exhibit three nematic phases, namely N, Nx and ferronematic NF. N is found to be a conventional nematic but shows two additional unusual features: the presence of optically active domains and a linear electro optic response to the applied signal. The optical activity is explained by a segregation of the stable helical conformers of the opposite sense; these preferring to form domains with an optical rotation of ±4°/μm. This is the first example of helical segregation observed in non-chiral molecules in the high temperature nematic phase. Unlike the ordinary nematic liquid crystalline material, this compound shows a linear electro optical response to the applied electric field, thereby confirming the polar nature of this phase. NF is the ferroelectric nematic as reported previously. The strong polar azimuthal surface interaction energy in the NF phase stabilises a homogeneous structure in planar-aligned cells that are rubbed parallel and gives a twisted structure in cells rubbed antiparallel. [ABSTRACT FROM AUTHOR]

Experimental limitations such as design complexity and low optical throughput have prevented photonic nanojet (PNJ) and photonic hook (PH) measurements from demonstrating and characterizing the implementation of narrow intense electromagnetic beams generated from dielectric microelements with circular symmetry. Near-fields optical microscopy can mitigate these limitations and still present a capability of detecting a highly localized electromagnetic beam for applications in step-index media. Here we model a localized PNJ and PH formation in step-index media. We show that despite negligible refractive index contrast between the water ( n w a t e r = 1.33 ) and silica microcylinder (∼1.1), a formation of PNJ and PH is observed with equivalent performance compared to that of silica microcylinder embedded in air ( n a i r = 1 ). This model features a practical fiber source and silica microcylinder as an auxiliary structure. Simultaneously, we performed experimental characterization of a photonic nanojet generated from an optical fiber and studied the resulting near-fields. Our electromagnetic simulation results are in good agreement with the experimental ones, demonstrating a full width at half maximum (FHWM) with a relative error of 0.64%. This system will make fiber-based nanojet realization and characterization accessible and practical for optics and laser engineering applications, super-resolution imaging, and nanolithography. [ABSTRACT FROM AUTHOR]

Over- or under-expression of erythropoietin-production human hepatocellular receptors (Eph) and their ligands are associated with various diseases. Therefore, these molecular biomarkers can potentially be used as binding targets for the delivery of therapeutic and/or imaging agents to cells characterized by such irregular expressions. We have engineered nanoparticles derived from erythrocytes and doped with the near-infrared (NIR) FDA-approved dye, indocyanine green. We refer to these nanoparticles as NIR erythrocyte-derived transducers (NETs). We functionalized the NETs with the ligand-binding domain of a particular Eph receptor, EphB1, to target the genetically modified human dermal microvascular endothelial cells (hDMVECs) with coexpression of EphB1 receptor and its ligand ephrin-B2. This cell model mimics the pathological phenotypes of lesional endothelial cells (ECs) in port wine stains (PWSs). Our quantitative fluorescence imaging results demonstrate that such functionalized NETs bind to the ephrin-B2 ligands on these hDMVECs in a dose-dependent manner that varies sigmoidally with the number density of the particles. These nanoparticles may potentially serve as agents to target PWS lesional ECs and other diseases characterized with over-expression of Eph receptors or their associated ligands to mediate phototherapy.

In infantile abusive head injury (AHT), subdural haemorrhage (SDH) is commonly held to result from traumatic damage to bridging veins traversing from the surface of the brain to the dura and dural venous sinuses. However, there are limited published radiological or autopsy demonstrations of ruptured bridging veins and several authors also assert that bridging veins are too large to rupture due to the forces associated with AHT. There have been several studies on the size, locations and numbers of adult bridging veins and there is one small study of infant bridging veins. However, there are no microscopic studies of infant bridging veins and only a select few ultrastructural investigations of adult bridging veins. Hitherto, it has been assumed that bridging veins from infants and younger children will display the same anatomical characteristics as those in adulthood. At 19 neonatal, infant and young child post-mortem examinations, we macroscopically examined and sampled bridging veins for microscopy. We compared the histology of those samples with bridging veins from an older child and two adults. We demonstrate that adult bridging veins are usually surrounded by supportive meningeal tissue that appears to be lacking or minimally present around the bridging veins of younger children. Neonatal, infant and young children's veins had a free 'bridging' section. Neonatal and infant bridging veins had smaller diameter ranges and thinner walls (some only 5–7 µm) than those seen in older children and adults. Bridging vein walls contained both fine strands of elastic fibers and a more pronounced elastic lamina. The presence of an elastic lamina occurred more frequently in the older age groups These anatomical differences between the veins of adults and young children may help to explain apparent increased vulnerability of neonatal/infant bridging veins to the forces associated with a shaking-type traumatic event. [ABSTRACT FROM AUTHOR]

Ultra-fast optical pulses are the most ephemeral sensing paradigm ever devised, examining events over incredibly brief timescales with broadband illumination. A consequence of sensing at timescales lower than a picosecond is that pulse characterization cannot be done with traditional analog-to-digital samplers and must be ascertained from integrating intensity sensors. Techniques for pulse characterization have been constructed using combinations of time-invariant and time-variant filter responses to create non-linear but invertible intensity datasets (Walmsley & Dorrer, 2009). In this paper, we develop a novel high-order phase retrieval technique to perform pulse characterization from a single-pixel integrating sensor measuring integrated intensity of auto-convolution (IIAC). We examine gradient descent's ability to recover signals as a function of signal dimension and measurement count, and we demonstrate the effective use of iterative hard tensor thresholding as an initializer. Finally, we demonstrate IIAC recovery in a laboratory setting to recover the time profile of a complex laser pulse. We assert that the IIAC recovery solution demonstrated here simultaneously provides the optics community with a pulse characterization technique that scales to low-power microscopy systems and provides the optimization community with a physically motivated high-order phase retrieval problem enhanced by low-rank tensor processing. • Ultrafast pulse characterization is built largely on auto-correlation datasets. • Tensor iterative hard thresholding and gradient techniques demonstrate algorithmic superiority. • These methods are applied to a condensed measurement not previously studied in optics or optimization. • New measurement type relieves requirement in optical systems for spectral or 2D sensor. • Wirtinger gradient descent and tensor IHT are used to effectively invert new data type in simulation and lab data. [ABSTRACT FROM AUTHOR]

We are combining two optical techniques, pulsed photothermal radiometry (PPTR) and diffuse reflectance spectroscopy (DRS), for noninvasive assessment of the structure and composition of human skin in vivo. The analysis involves simultaneous multidimensional fitting of the measured PPTR signals and DRS spectra with predictions of a numerical model of light transport (Monte Carlo) in a four-layer model optical model of human skin, accounting for the epidermis, papillary and reticular dermis, and subcutis. The assessed epidermal thickness values were tested by coregistration with a multiphoton microscope, which provides vertical sectioning capability based on two-photon excited fluorescence and second-harmonic generation in selected skin components. The comparison shows that these values correspond well to the maximal epidermal thicknesses measured in the multiphoton microscopy images, the rete ridges.

Volumetric imaging tools that are simple to adopt, flexible, and robust are in high demand in the field of neuroscience, where the ability to image neurons and their networks with high spatiotemporal resolution is essential. Using an axially elongated focus approximating a Bessel beam, in combination with two-photon fluorescence microscopy, has proven successful at such an endeavor. Here, we demonstrate three-photon fluorescence imaging with an axially extended Bessel focus. We use an axicon-based module that allowed for the generation of Bessel foci of varying numerical apertures and axial lengths, and apply this volumetric imaging tool to image mouse brain slices and for in vivo imaging of the mouse brain.

Most flying-spot optical coherence tomography and optical coherence microscopy (OCM) systems use a symmetric confocal geometry, where the detection path retraces the illumination path starting from and ending with the spatial mode of a single-mode optical fiber. Here we describe a visible light OCM instrument that breaks this symmetry to improve transverse resolution without sacrificing collection efficiency in scattering tissue. This was achieved by overfilling a water immersion objective on the illumination path while maintaining a conventional Gaussian mode detection path (1/e2 intensity diameter ∼0.82 Airy disks), enabling ∼1.1  μm full width at half-maximum (FWHM) transverse resolution. At the same time, a ∼0.9  μm FWHM axial resolution in tissue, achieved by a broadband visible light source, enabled femtoliter volume resolution. We characterized this instrument according to paraxial coherent microscopy theory and, finally, used it to image the meningeal layers, intravascular red blood cell-free layer, and myelinated axons in the mouse neocortex in vivo through the thinned skull.

Inline inspection is becoming an essential tool for industrial high-quality production. Unfortunately, the desired acquisition speeds and needs for high-precision imaging are often at the limit of what is physically possible, such as a large field of view at a high spatial resolution. In this paper, a novel light-field and photometry system is presented that addresses this trade off by combining microscopic imaging with special projection optics to generate a parallax effect. This inline microscopic system, together with an image processing pipeline, delivers high-resolution 3D images at high speeds, by using a lateral transport stage changing the optical perspective. Scanning speeds of up to 12 mm/s can be achieved at a depth resolution of 2.8 μm and a lateral sampling of 700 nm/pixel, suitable for inspection in high-quality manufacturing industry. [ABSTRACT FROM AUTHOR]