Front Cover: A high‐throughput all‐optical laser‐scanning imaging flow cytometer with biomolecular specificity and subcellular resolution (J. Biophotonics 2/2018)

A new type of high‐throughput imaging flow cytometer (>20 000 cells s^‐1) based upon an all‐optical ultrafast laser‐scanning imaging technique, called free‐space angular‐chirp‐enhanced delay (FACED) is reported. FACED imaging flow cytometers enables high‐throughput visualization of functional morphology of individual cells with subcellular resolution. It critically empowers largescale and deep characterization of single cells and their heterogeneity with high statistical power— an ability to become increasingly critical in single‐cell analysis adopted in a wide range of biomedical and life‐science applications. Further details can be found in the article by Wenwei Yan et al. ( e201700178 )

     

Front Cover: A theoretical‐experimental methodology for assessing the sensitivity of biomedical spectral imaging platforms,assays, and analysis methods (J. Biophotonics 1/2018)

A hyperspectral image data cube acquired from HEK‐293 cells labeled with cytoplasmic and nuclear stains: Calcein Green and NucBlu. The top view (XY plane) displays three spectrally unmixed channels for cellular autofluorescence (red), Calcein Green (green), and NucBlue (blue). The Z axis shows spectral information, from low to high wavelength. The article by Leavesley and colleagues describes an approach for calculating the sensitivity of spectral imaging assays for detecting a fluorescence signature within a mix of other signatures or autofluorescence. Further details can be found in the article by Silas J. Leavesley et al. ( e201600227 ).

     

Back Cover: Non‐invasive optical method for real‐time assessment of intracorneal riboflavin concentration and efficacy of corneal cross‐linking (J. Biophotonics 7/2018)

We disclose a theranostic device for performing image‐guided riboflavin/UV‐A corneal cross‐linking. The device determines treatment efficacy by real time monitoring of riboflavin concentration in the corneal stroma. The study shows efficacy of the device in eye bank human donor tissues. Further details can be found in the article by Giuseppe Lombardo et al. ( e201800028 )

     

Inside Front Cover: Enhanced volumetric imaging in 2‐photon microscopy via acoustic lens beam shaping (J. Biophotonics 2/2018)

Two‐photon microscopy is the tool of choice for fluorescence imaging of deep tissues with high resolution, but can be limited in three‐dimensional acquisition speed and penetration depth. In this work, these issues are addressed by using an acoustic optofluidic lens capable of ultrafast beam shaping on a pixel basis. Driving the lens with different phase profiles enables high‐speed volumetric imaging, or enhanced signal‐to‐background for deeper penetration. Further details can be found in the article by Simonluca Piazza et al. ( e201700050 )

     

Inside Cover: Fiber‐based fluorescence lifetime imaging of recellularization processes on vascular tissue constructs (J. Biophotonics 9/2018)

Tissue autofluorescence provides fluorescence lifetime contrast between acellular tissue and that containing newly seeded cells. Fiber‐based fluorescence lifetime imaging (FLIm) can be used for tracking recellularization of engineered vascular grafts and potential matrix remodeling at large scale, without compromising sample integrity. FLIm cellular contrast was verified in a subset of samples seeded with eGFP‐labelled cells. Results suggests fiberbased FLIm is a suitable tool for monitoring recellularization of engineered tissue nondestructively. Further details can be found in the article by Alba Alfonso‐Garcia, Jeny Shklover, Benjamin E. Sherlock, et al. ( e201700391 ).

     

Front Cover: Dual‐wavelength hybrid optoacoustic‐ultrasound biomicroscopy for functional imaging of large‐scale cerebral vascular networks (J. Biophotonics 9/2018)

We present a hybrid dual‐wavelength optoacoustic and ultrasound bio‐microscope capable of rapid transcranial visualization of morphology and oxygenation status of large‐scale cerebral vascular networks. Imaging of entire cortical vasculature in mice is achieved with single capillary resolution and complemented by simultaneously acquired pulse‐echo ultrasound microscopy scans of the mouse skull. The new approach holds potential to facilitate studies into neurological and vascular abnormalities of the brain. Further details can be found in the article by Johannes Rebling, Héctor Estrada, Sven Gottschalk, et al. ( e201800057 ).

     

Inside Back Cover: The conformation of bovine serum albumin adsorbed to the surface of single all‐dielectric nanoparticles following light‐induced heating (J. Biophotonics 7/2018)

Germanium vs Silicon: All‐dielectric nanoparticles provides the heat resistance for proteins under light‐induced heating. Further details can be found in the article by Andrei A. Krasilin et al. ( e201700322 )

     

Inside Back Cover: In vivo full‐field functional optical hemocytometer (J. Biophotonics 2/2018)

Full‐field functional optical hemocytometer (FFOH), based on the absorption intensity fluctuation modulation (AIFM) effect, is in vivo label‐free image method for capillaries of near‐transparent live biological specimens. FFOH can provide a flow video, flow velocity measurement and RBC count, simultaneously. The zebrafish experimental result shows the potential to study the physiological mechanisms of the blood circulation systems. Further details can be found in the article by Fuli Zhang et al. ( e201700039 )

     

Inside Back Cover: Development of a 3‐dimensional tissue lung phantom of a preterm infant for optical measurements of oxygen—Laser‐detector position considerations (J. Biophotonics 3/2018)

The picture depicts the different 3d‐printed organs, thorax, lungs, heart and bone. Assembled it is used as an optical phantom of a preterm infant for performing percutaneous optical measurements of the gas content in the lungs. In order to simulate the optical properties of the tissue, the heart and thorax can be filled with liquid phantoms, a mixture of Intralipid and Indian Ink. Further details can be found in the article by Jim Larsson et al. ( e201700097 ).

     

Back Cover: In‐vitro analysis of early calcification in aortic valvular interstitial cells using Laser‐Induced Breakdown Spectroscopy (LIBS) (J. Biophotonics 1/2018)

Quantitative laser‐induced breakdown spectroscopy (LIBS) is successfully used for in‐vitro analysis of early stage calcification in aortic valvular interstitial cells (VICs). LIBS results indicate 5‐fold improvement in the detection limit of calcium deposition in VICs over cell histology techniques involving staining and colorimetric calcium assays. These results can establish LIBS at the forefront of early detection of calcification in VICs for pathological studies on Calcific Aortic Valve Disease (CAVD). Further details can be found in the article by Seyyed Ali Davari et al. ( e201600288 ).

     

A compact fiber‐optic probe‐based singlet oxygen luminescence detection system

This paper presents a novel compact fiberoptic based singlet oxygen near‐infrared luminescence probe coupled to an InGaAs/InP single photon avalanche diode (SPAD) detector. Patterned time gating of the single‐photon detector is used to limit unwanted dark counts and eliminate the strong photosensitizer luminescence background. Singlet oxygen luminescence detection at 1270 nm is confirmed through spectral filtering and lifetime fitting for Rose Bengal in water, and Photofrin in methanol as model photosensitizers. The overall performance, measured by the signal‐to‐noise ratio, improves by a factor of 50 over a previous system that used a fiberoptic‐coupled superconducting nanowire single‐photon detector. The effect of adding light scattering to the photosensitizer is also examined as a first step towards applications in tissue in vivo.

     

Monochromatic subdiffusive spatial frequency domain imaging provides in‐situ sensitivity to intratumoral morphological heterogeneity in a murine model

For the first time, spatially resolved quantitative metrics of light scattering recovered with sub‐diffusive spatial frequency domain imaging (sd‐SFDI) are shown to be sensitive to changes in intratumoral morphology and viability by direct comparison to histopathological analysis. Two freshly excised subcutaneous murine tumor cross‐sections were measured with sd‐SFDI, and recovered optical scatter parameter maps were co‐registered to whole mount histology. Unique clustering of the optical scatter parameters vs. γ (i.e. diffuse scattering vs. relative backscattering) evaluated at a single wavelength showed complete separation between regions of viable tumor, aggresive tumor with stromal growth, varying levels of necrotic tumor, and also peritumor muscle. The results suggest that with further technical development, sd‐SFDI may represent a non‐destructive screening tool for analysis of excised tissue or a non‐invasive approach to investigate suspicious lesions without the need for exogenous labels or spectrally resolved imaging.

     

Inside Front Cover: Near‐infrared bone densitometry: A feasibility study on distal radius measurement (J. Biophotonics 7/2018)

This study provides a simple method to detect human distal radius bone density based on near infrared (NIR) imaging. The information of bone mineral density can be measured by transluminational optical bone densitometric system. Compared to dual‐energy x‐ray absorptiometry (DXA) results in clinical trial, NIR images show a strong correlation to DXA. Further details can be found in the article by Chun Chung, Yu‐Pin Chen, Tsai‐Hsueh Leu, and Chia‐Wei Sun ( e201700342 ).

     

Three‐dimensional mapping of the orientation of collagen corneal lamellae in healthy and keratoconic human corneas using SHG microscopy

Keratoconus is an eye disorder that causes the cornea to take an abnormal conical shape, thus impairing its refractive functions and causing blindness. The late diagnosis of keratoconus is among the principal reasons for corneal surgical transplantation. This pathology is characterized by a reduced corneal stiffness in the region immediately below Bowman's membrane, probably due to a different lamellar organization, as suggested by previous studies. Here, the lamellar organization in this corneal region is characterized in three dimensions by means of second‐harmonic generation (SHG) microscopy. In particular, a method based on a three‐dimensional correlation analysis allows to probe the orientation of sutural lamellae close to the Bowman's membrane, finding statistical differences between healthy and keratoconic samples. This method is demonstrated also in combination with an epi‐detection scheme, paving the way for a potential clinical ophthalmic application of SHG microscopy for the early diagnosis of keratoconus.

SHG image acquired with sagittal optical sectioning ( A ) of a healthy cornea and ( B ) of a keratoconic cornea. Scale bars: 30 μm.     

Inside Cover: Noninvasive,high‐speed,near‐infrared imaging of the biomolecular distribution and molecular mechanism of embryonic development in fertilized fish eggs (J. Biophotonics 4/2018)

The biomaterial distribution and its molecular mechanism of embryonic development in Japanese medaka fish were visualized without staining using high‐speed near‐infrared imaging. It was a remarkable achievement to visualize the structures of eyes, lipid bilayer membranes, micelles, and water structural variations at the interface of different substances. Furthermore, insights on lipid metabolism and membrane functions were obtained from the biased distribution of lipoproteins and the presence of unsaturated fatty acids in the egg membrane. Further details can be found in the article by Mika Ishigaki ( e201700115 )

     

Cancer cell identification by bi‐color ZnO and TiO2 nanowires

Semiconductor nanocomposites provide advantages beyond the capability of typical fluorescent materials for cancer detection. In this work, nanowire‐based probes with dual color channels are employed to demonstrate the capacity of cancer cell detection. Purple emitting ZnO/antibody probes are applied to detect cancer cells and meanwhile TiO₂/antibody probes with green light emission are applied to identify normal fibroblast cells. A series of quantitative analyses are conducted to verify the correlation between the concentrations of ZnO and TiO₂ probes, cell numbers, and peak intensities of the PL spectra. The results provide a quantitative reference for developing nanowire‐based cancel cell probes.

     

Back Cover: Autofluorescence and white light imaging‐guided endoscopic Raman and diffuse reflectance spectroscopy for in vivo nasopharyngeal cancer detection (J. Biophotonics 4/2018)

An integrated 4‐modality endoscopy system combining white light imaging, autofluorescence imaging, diffuse reflectance spectroscopy and Raman spectroscopy technologies was developed for in vivo endoscopic nasopharyngeal cancer detection. Both high diagnostic sensitivity (98.6%) and high specificity (95.1%) for differentiating cancer from normal tissue sites were achieved using this system combined with multivariate diagnostic algorithm, demonstrating great potential for improving real‐time, in vivo diagnosis of cancer at endoscopy. Further details can be found in the article by Duo Lin et al. ( e201700251 )

     

Analysis of major elements in pigmented melanocytic chicken skin using laser‐induced breakdown spectroscopy

The concentration difference of major elements in melanocytic skin with respect to pigmentation level is analysed by laser‐induced breakdown spectroscopy (LIBS) to investigate the applicability of LIBS as an in situ feedback tool for selective and complete laser removal of melanocytic skin tissue like nevus. The skin of black silkie chicken which had a characteristic darkly pigmented perifollicular skin surrounded by lightly pigmented extrafollicular skin was used as the sample. The results showed higher LIBS signal intensities of Ca²⁺ and Mg²⁺ but lower intensities of Na⁺, Cl^– and K⁺ in the perifollicular skin than in the extrafollicular skin, which demonstrated the feasibility to use LIBS as a reliable method to distinguish skin tissues with difference in pigmentation level.

Plasma emission of biochemical elements generated with a laser irradiation on melanocytic skin lesion.     

Bessel‐beam illumination in dual‐axis confocal microscopy mitigates resolution degradation caused by refractive heterogeneities

One of the main challenges for laser‐scanning microscopy of biological tissues with refractive heterogeneities is the degradation in spatial resolution that occurs as a result of beam steering and distortion. This challenge is particularly significant for dual‐axis confocal (DAC) microscopy, which achieves improved spatial‐filtering and optical‐sectioning performance over traditional confocal microscopy through off‐axis illumination and collection of light with low‐numerical aperture (NA) beams that must intersect precisely at their foci within tissues. DAC microscope image quality is sensitive to positional changes and distortions of these illumination‐ and collection‐beam foci. Previous studies have shown that Bessel beams display improved positional stability and beam quality than Gaussian beams when propagating through tissues with refractive heterogeneities, which suggests that Bessel‐beam illumination may enhance DAC microscopy of such tissues. Here, we utilize both Gaussian and Bessel illumination in a point‐scanned DAC microscope and quantify the resultant degradation in resolution when imaging within heterogeneous optical phantoms and fresh tissues. Results indicate that DAC microscopy with Bessel illumination exhibits reduced resolution degradation from microscopic tissue heterogeneities compared to DAC microscopy with conventional Gaussian illumination.

     

Assessment of ALA‐induced PpIX production in porcine skin pretreated with microneedles

Photodynamic therapy (PDT) is used for skin treatments of premalignant and cancer lesions and recognized as a non‐invasive technique that combines tissue photosensitization and subsequent exposure to light to induce cell death. However, it is limited to the treatment of superficial lesions, mainly due to the low cream penetration. Therefore, the improvement of transdermal distribution of aminolevulinic acid (ALA) is needed. In this study, the kinetics and homogeneity of production of ALA‐induced PpIX after the skin pre‐treatment with microneedles rollers of 0.5, 1.0 and 1.5 mm length were investigated. An improvement in homogeneity and production of PpIX was shown in a porcine model.

Widefield fluorescence imaging three hours after the topical application of ALA‐cream in the combined treatment with microeedles rollers.