This PDF file contains the front matter associated with SPIE Proceedings Volume 11922, including the Title Page, Copyright information, and Table of Contents.

We propose the combination of squared cubic phase profiles and spherical aberration patterns to further improve the quality of multiphoton microscopy images. The benefit will be analyzed through the use of objective image quality metrics.

We present a microscopy method capable of measuring aberrations in all the poits of the field of view and to correct for the field-dependent aberrations in a closed loop multi conjugated AO system using two deformable lenses and no wavefront sensor.

We demonstrate focus optimisation through multimode optical fibre using sensorless adaptive optics. The optimisation can correct for the three-dimensional shift occurring when re-positioning the fibre into the system for chronic brain imaging.

We present a robust, low-cost neural network-based optical autofocus system that can operate over a range of ±100μm with submicron precision, enabling automated high-content super-resolved imaging with a 1.3 NA objective lens.

We applied an unmatched back projector based deconvolution algorithm to reduce image reconstruction time in light field microscopy. 3D laser written microstructures were fabricated and used to characterize the light field imaging.

Holographic endoscopes with sub-millimeter diameter require the compensation of inter-core phase distortions, which is accomplished often with an SLM. We present the use of 3D printed DOE as a robust integrative technique for biomedicine.

Far-field optical microscopy typically suffers from limited resolution, speed and imaging depth. Endoscopic imaging via a multimode fiber combined with wavefront shaping and computational reconstruction offers imaging beyond the Abbe and Nyquist limits.

Fiber-based endoscopic compressive imaging is a promising field for minimal invasive, in vivo imaging below the diffraction limit and enhanced acquisition time. Here, we analyze and present the theoretical limits of speckle-based compressive imaging.

A fiber coupled, sub ps Ti:sapphire laser suitable for in vivo, stain free, 3D imaging of skin alterations is introduced. It is pumped by a low cost, 2.1 W pump laser and delivers 0.6 1 ps high peak power pulses optimized for fiber delivery.

We report wide-field, epi-illumination microscopic imaging of biological tissues using single-shot random laser illumination. The random laser provides high-resolution, artefact-free images with better contrast and sharpness of structural details.

A fluorescence correlation contrast is a straightforward approach to super-resolution imaging. Combining a SPAD array with a novel detection scheme (ISM), we obtain images with up to x4 times resolution enhancement.

Multispectral fluorescence lifetime imaging microscopy (λFLIM) is a high sensitivity technique for multifold applications. We present a λFLIM system and a compressive sensing data acquisition strategy, which allows one to reduce the measurement time.

Quantitative FRET imaging in living cells is made possible on a simple fluorescence microscope with this new method. The single-step calibration is robust and gives absolute FRET values, independent of the instrument.

We quantify multi-labelled streptavidin on a surface with un-known distribution of fluorophores in situ using correlation spectroscopy and photobleaching. This will allow the characterization of the functionalization of biomaterials for future regenerative medicine applications.

Ultra-broadband lasers allow simultaneous excitation of various dyes and endogenous markers, used for metabolic imaging during treatment of live-cells with anticancer drugs. The versatility of few cycle lasers for biomedical imaging is demonstrated.

We present a new implementation of adaptive optics for light-sheet microscopy, with a direct extended-scene wavefront sensing measurement for fast aberration correction. We report AO-enhanced images of GCaMP in freshly dissected drosophila brains.

We propose a framework to quantify photodamage in multiphoton light-sheet microscopy. Using cardiac imaging in live zebrafish embryos, we demonstrate an order of magnitude signal enhancement is safely obtained by adjusting the laser repetition rate.

We present a single-shot volumetric imaging method, utilizing optical projection tomography. We record projections simultaneously, implementing compressive sensing and machine learning to record up to 70 (camera limited) 1x1x1.9mm volumes/second.

We explore wavefront shaping techniques to produce and control illumination patterns for use on Light-Sheet Fluorescence Microcopy (LSFM). We demonstrate the use of these patterns to perform in vivo imaging using fluorescently labeled specimens.

We implement polarization-resolved second harmonic generation microscopy to characterize the orientation distribution of collagen lamellae in human cornea. We evidence a less ordered distribution in keratoconic corneas, in agreement with their deteriorated mechanical behavior.

Wide-field Polarization-resolved Second-Harmonic Generation microscopy is a label-free imaging technique which highlights molecular organization of collagenous tissues, enabling high-throughput quantitative biomedical imaging and cancer diagnostics.

Nonlinear response of complex chiral structures depends on the fibers configuration in the focal volume. The impact of various configuration on nonlinear parameters extracted using a polarimetric second harmonic generation (SHG) microscope has been elucidated.

We present a novel nonlinear microscopy modality using a time-domain ptychographic phase measurement, i²PIE, to compress 80 MHz supercontinuum pulses from an ANDi PCF used as excitation source, improving contrast at reduced average power.

In-vivo tracking based on harmonic nanoparticles is so far not exploited because of a lack on an appropriate tool. Here we present a new approach based on there-design of the laser space parameters.

Individual small gold nanoparticles are imaged in 3D background-free with high contrast by four-wave-mixing interferometry inside living mouse oocytes and multicellular organs, despite the strong linear scattering background in these large samples.

A hyperspectral photoacoustic remote sensing microscope is used to investigate and image optical absorption contrast in live and resected chicken tissues ranging from 250 nm to 1210 nm highlighting DNA, hemeproteins, and lipids.

We demonstrate the use of Photoacoustic Remote Sensing (PARS) and scattering microscopy capable of acquiring virtual depth-resolved images of tissues with virtual contrast of hematoxylin using PARS & eosin using scattering microscopy.

Polarized light microscopy can be used to objectively quantify tumour-stroma ratio (TSR), a prognostic indicator validated in several cancers. Polarimetric results correctly differentiated between high- and low-stroma samples, and future work holds promise for improved separation.

We evaluate the signal-to-noise sensitivity of distance measures commonly used in the hierarchical cluster analysis of bacterial Raman spectra and discuss their applicability for analyzing the datasets generated in Raman spectral imaging.

We demonstrate a combination of polarization-resolved with fluorescence optical scanning microscopy that offers polarizing based information along with molecular view of sample as a versatile tool for imaging the chromatin organization.

Refractive index tomography compatible with conventional microscope is used for analyzing primary rat hepatocytes injury induced by pharmacological treatments. We found that mitochondria malfunctioning is correlated with refractive index variation.

We introduce lattice light-sheet system with an incoherent detection module that scans an extended FOV of 208x208 μm². We demonstrated the IHLLS 3D imaging capabilities, amplitude and phase distributions, of polystyrene beads and nerve cells.

We present a robust, low-cost single-shot implementation of differential phase microscopy utilising a polarisation-sensitive camera to simultaneously acquire 4 images from which the phase gradients and quantitative phase image can be calculated.

A lateral shearing interferometer is used to take holographic images and videos of red blood cells (RBCs). The extracted physical and mechanical parameters can be used to define the state-of-health of the RBCs.

Multifrequency AOTF operation provides accurate controlling of the transmitted optical spatial frequencies. We demonstrate its advantages for increased throughput in acousto-optic phase imaging and flexibility of controlling annular optical traps.

We apply pulsed optical phase contrast microscopy to measure the absolute pressure amplitudes of complex ultrasonic fields generated by planar and focused transducers at frequencies up to 20 MHz. 2021 National Institute of Standards and Technology.

Stimulated Raman scattering (SRS) microscopy is a well-established non-linear optical technique for label-free identification of biochemical components in cells and tissues. In this contribution, we present broadband SRS microscopy with differential multichannel-lock in detection

The extracellular matrix plays a crucial role in tumor development and efficacy of chemotherapy. We present a combined imaging approach to investigate and quantify the collagen architecture on the mm to μm scale.

We present two methods for the image-based quality control of cell spheroids used for drug development. The strengths and limitations of scanning laser optical tomography and two-photon excitation microscopy are evaluated and compared.

A compact non-contact multiphoton microscope for the imaging in-vivo anterior segment of the human eye is presented. Different recording settings are evaluated to obtain maximum stability and optimum image quality within ocular safety limits.

We present a classification scheme of different types of RBC in human blood using a Diffraction Phase Microscopy (DPM) setup combine with Machine learning. The experimental study demonstrated an improved accuracy in classification of RBCs.

In this work we investigated how changing the labeling construct of porcine liver tissue enhances labeling speed. Our results show that bi-directional labeling can indeed reduce labeling time with respect to the standard uni-directional labeling.

A novel imaging system is developed to overcome the challenge of paraffin contamination in fixed tissues. Single-shot wide-field micro-images are captured to construct large-scale collagen-maps for understanding tendon damage and healing processes.

We demonstrate the ability to perform fast ultraviolet photoacoustic remote sensing microscopy of tissue margins using a mosaic scanning approach where a camera-based system is used to determine scan coordinates within a 2-mm margin.

Optical sectioning technologies achieve high precision localization by reducing the background photon count. We use tilted light-sheet microscopy to achieve optical sectioning in localization microscopy, enabling thick sample observation and low background photon count images. A deformable mirror was incorporated to generate a tetrapod point spread function (PSF), enabling high resolution 3D localization. DNA-PAINT was imaged with 15 nm transverse and 60 nm axial resolution.

We propose HiLo based line-scanning temporal focusing microscopy to enhance contrast and axial confinement in deep imaging, and demonstrate its superiority by volumetric imaging of microglia and neurons in mouse brains in vivo.

Skin malignancies diagnostics based on spectral reflection data obtained from skin, are considered. The diagnostics method foresees application of acousto-optic tunable filters. The experimental data connected with the method realization, are listed.

Hyperspectral-imaging is label-free imaging technique. We designed hyperspectral source based on chromatic dispersion property of off-the-shelf lenses, that can be incorporated into standard endoscope/microscope to perform hyperspectral-imaging.

Low concentration Phloxine B staining was applied for nonlinear microscope mosaic imaging of skin alterations in Pseudoxanthoma Elasticum sections helping in determination of illness severity by quantitative analysis of calcium deposits in the skin.

Imaging cytometry is a vital tool in many bio-imaging studies. Here we introduce, Differentiable Microscopy (δμ) a general end-to-end optimizable design framework to improve the throughput of imaging cytometry using content aware sampling and reconstruction.

Fat in human milk forms the main energy source for infants and is the most variable component in terms of concentration and composition. Knowledge on changes in human milk lipid composition and conformational state during a single breastfeed contributes to an in-depth understanding of lipid synthesis in the mammary gland. Therefore, the objective of this study was to evaluate the differences in fatty acid length, degree of unsaturation (lipid composition) and lipid phase (lipid conformational state) of milk released at different stages during a breastfeed (fore-, bulk- and hindmilk). A total of 30 samples from 10 lactating subjects were investigated using confocal Raman spectroscopy. No significant differences in lipid composition were observed between fore-, bulk- and hindmilk samples, which is consistent with literature. A new finding from this study is that the lipid conformational state in human fore-, bulk- and hindmilk was significantly different at room temperature. The lipid phase of foremilk was almost crystalline and the lipid phase of hindmilk was almost liquid. Based on this observation, we hypothesize that lipid synthesis in the mammary gland changes during a single breastfeed.

Terahertz solid immersion microscopy is a imaging modality, allowing to overcome Abbe diffraction limit and obtain resolution of 0.15λ. For numerical modelling of solid immersion systems and various objects used finite-difference time-domain method.

Raman spectral imaging with cytochrome-c resonant excitation of a few hundred randomly chosen cells per culture allows assessing culture vitality of slow growing bacterial cultures with low cell counts based on a reliable spectral marker.

Lensless endoscopy based on multicore fibres is a promising concept for 3D imaging deep inside tissue. In-depth design studies regarding novel aperiodic multicore fibre designs and their advantages in terms of endoscopy performance are presented.

Liquid reference samples for 2-photon fluorescence lifetime investigations were developed, based on coumarin 1 and 6 and 4-hydroxy-TEMPO as quencher. Spectral and fluorescence lifetime characteristics similar to intracellular NADH and FAD were achieved.

We developed an innovative multimodal nonlinear microscope, that operates in four different modalities. Using this tool on murine models, we set procedures and protocols to understand the role of dipeptidyl peptidase 3 in skeletal diseases.

Accumulation of Radachlorin photosensitizer in 3T3 cells was evaluated using holographic tomography and fluorescence microscopy. Comparative analysis of images obtained by these techniques evidences drug accumulation in small intracellular structures located primarily in the juxtanuclear area.

In this paper, a transmission mode optical technique is required to achieve a super-resolved image. Basically, this technique is used for high resolution and for large field of view (FOV). The object is illuminated with deterministic mask patterns, and the object encoded information is transmitted to the sensor that is positioned at an optimum distance and records the images. A Spatial light modulator (SLM) is used as a mask and phase distribution of the SLM is changed ten times to captures ten different images. By using the angular spectrum process the captured images are backpropagated sequentially and reconstruct the super-resolved image. The use of multiple deterministic masks and Spatial Light Modulator (SLM) improved the quality of the reconstruction method.

Ptychographic imaging is centered on the phase retrieval of the sample from the diffraction patterns recorded at the sensor. Ptychography is a lens less computational imaging method that has various types. The single aperture ptychographic imaging technique is discussed in this design. Firstly, the simulation is done by MATLAB and after that the physical experiment is performed. The amplitude and phase-reconstructed images are retrieved by pinhole illumination. The sample is illuminated through the pinhole (generated by SLM) and the encoded sample information propagates to the CMOS, which is positioned at an optimum distance and captures the images. The Ten different diffraction patterns of the specimen are recorded by the CMOS sensor by shifting it axially and the data is recorded on the computer. To retrieve the phase and amplitude information about sample from raw images the reconstruction process is applied through MATLAB. The recorded field is back propagated to the object plane in the reconstruction process. Finally, by including all the back propagated images, a final reconstructed image with higher resolution is obtained using the stochastic gradient descent technique. The noise suppression approach is added to the final image to further increase resolution. The final image is highly resolved. Lens-less microscopy generates a cost-effective, portable optical device. Ptychography have many applications in medical field. It is used within the human body to detect biological cells. This provides good results in diagnosis of diseases.

We present the design and characterization of a 3D-printed spiral actuator towards focus adjustment in endoscopes. The actuator, holding an aspherical lens, can be electromagnetically actuated to achieve ~0.5 mm displacement at 10 mW power.