Deep learning-based classification of dermatological lesions given a limited amount of labelled data.

BACKGROUND: Artificial intelligence (AI) techniques are promising in early diagnosis of skin diseases. However, a precondition for their success is the access to large-scaled annotated data. Until now, obtaining this data has only been feasible with very high personnel and financial resources. OBJECTIVES: The aim of this study was to overcome the obstacle caused by the scarcity of labelled data. METHODS: To simulate the scenario of label shortage, we discarded a proportion of labels of the training set. The training set consisted of both labelled and unlabelled images. We then leveraged a self-supervised learning technique to pretrain the AI model on the unlabelled images. Next, we fine-tuned the pretrained model on the labelled images. RESULTS: When the images in the training dataset were fully labelled, the self-supervised pretrained model achieved 95.7% of accuracy, 91.7% of precision and 90.7% of sensitivity. When only 10% of the data were labelled, the model could still yield 87.7% of accuracy, 81.7% of precision and 68.6% of sensitivity. In addition, we also empirically verified that the AI model and dermatologists are consistent in visually inspecting the skin images. CONCLUSIONS: The experimental results demonstrate the great potential of the self-supervised learning in alleviating the scarcity of annotated data.

Risks and benefits of dermatological machine learning health care applications-an overview and ethical analysis.

BACKGROUND: Visual data are particularly amenable for machine learning techniques. With clinical photography established for skin surveillance and documentation purposes as well as progress checks, dermatology is an ideal field for the development and application of emerging machine learning health care applications (ML-HCAs). To date, several ML-HCAs have detected malignant skin lesions on par with experts or found overlooked visual patterns that correlate with certain dermatological diseases. However, it is well established that ML-HCAs come with ethical and social implications. OBJECTIVES: Currently, there is a lack of research that establishes model design, training, usage and regulation of such technologies sufficient to ensure ethically and socially responsible development and clinical translation, specifically within the field of dermatology. With this paper, we aim to give an overview of currently discussed ethical issues relating to dermatological ML-HCAs. METHODS: On the basis of a thematic, keyword-based literature search, we performed an ethical analysis against established frameworks of biomedical ethics. We combined our results with current, relevant normative machine learning ethics literature to identify the status quo of the ethics of ML-HCAs in dermatology. We describe the benefits and risks of dermatological ML-HCAs that are currently being developed for clinical purposes. RESULTS: The potential benefits range from better patient outcomes to better knowledge accessibility to decreasing health care disparities, that is, standards of care between different population groups. The risks associated with ML-HCAs range from confidentiality issues to individual patient outcomes as well as the exacerbation of prevalent health care disparities. We discuss the practical implications for all stages of dermatological ML-HCA development. CONCLUSION: We found that ML-HCAs present stakeholder-specific risks for patients, health care professionals and society, which need to be considered separately. The discipline lacks sufficient biomedical ethics research that could standardize the approach to ML-HCA model design, training, use and regulation of such technologies.

Spectral cross-cumulants for multicolor super-resolved SOFI imaging.

Super-resolution optical fluctuation imaging provides a resolution beyond the diffraction limit by analysing stochastic fluorescence fluctuations with higher-order statistics. Using nth order spatio-temporal cross-cumulants the spatial resolution and the sampling can be increased up to n-fold in all spatial dimensions. In this study, we extend the cumulant analysis into the spectral domain and propose a multicolor super-resolution scheme. The simultaneous acquisition of two spectral channels followed by spectral cross-cumulant analysis and unmixing increases the spectral sampling. The number of discriminable fluorophore species is thus not limited to the number of physical detection channels. Using two color channels, we demonstrate spectral unmixing of three fluorophore species in simulations and experiments in fixed and live cells. Based on an eigenvalue/vector analysis, we propose a scheme for an optimized spectral filter choice. Overall, our methodology provides a route for easy-to-implement multicolor sub-diffraction imaging using standard microscopes while conserving the spatial super-resolution property.

Erratum: Reduction of Abeta amyloid pathology in APPPS1 transgenic mice in the absence of gut microbiota.

This corrects the article DOI: 10.1038/srep41802.

Design of Acquisition Schemes and Setup Geometry for Anisotropic X-ray Dark-Field Tomography (AXDT).

Anisotropic X-ray Dark-field Tomography (AXDT) is a new imaging technique for reconstructing the three-dimensional scattering profile within a sample using the dark-field signal measured in an X-ray grating interferometry setup. As in any tomographic measurement, the acquisition geometry plays a key role in the accurate reconstruction of the scattering information. More- over, the anisotropic nature of the dark-field signal poses additional challenges for designing the acquisition protocols. In this work, we present an efficient approach to measure scattering orientations spread over the unit sphere and prove its efficacy using the knowledge from conventional tomography. In addition, we conclude (using analytical and experimental results) that placing the gratings such that the grating bars make an angle of 45 degrees with respect to the vertical direction is the optimal setup configuration for AXDT.

Reduction of Abeta amyloid pathology in APPPS1 transgenic mice in the absence of gut microbiota.

Alzheimer's disease is the most common form of dementia in the western world, however there is no cure available for this devastating neurodegenerative disorder. Despite clinical and experimental evidence implicating the intestinal microbiota in a number of brain disorders, its impact on Alzheimer's disease is not known. To this end we sequenced bacterial 16S rRNA from fecal samples of Aß precursor protein (APP) transgenic mouse model and found a remarkable shift in the gut microbiota as compared to non-transgenic wild-type mice. Subsequently we generated germ-free APP transgenic mice and found a drastic reduction of cerebral Aß amyloid pathology when compared to control mice with intestinal microbiota. Importantly, colonization of germ-free APP transgenic mice with microbiota from conventionally-raised APP transgenic mice increased cerebral Aß pathology, while colonization with microbiota from wild-type mice was less effective in increasing cerebral Aß levels. Our results indicate a microbial involvement in the development of Abeta amyloid pathology, and suggest that microbiota may contribute to the development of neurodegenerative diseases.

Anisotropic X-Ray Dark-Field Tomography: A Continuous Model and its Discretization.

The x-ray dark-field signal measured in grating interferometers is anisotropic, depending on both the beam direction and the grating orientation with respect to the sample. We present a novel general closed-form, continuous forward model of the anisotropic dark-field signal. Furthermore, we derive a discretization using spherical harmonics, leading to a large-scale linear inverse problem. We present first experimental results of a wooden sample, demonstrating marked advantages over previous results, in particular, the resolution of multiple scattering directions in one volume element.

Towards high resolution optical imaging of beta cells in vivo.

Endocrine beta cells produce and release insulin in order to tightly regulate glucose homeostasis and prevent metabolic pathologies such as Diabetes Mellitus. Optical imaging has contributed greatly to our current understanding of beta cell structure and function. In vitro microscopy of beta cell lines has revealed the localization of molecular components in the cell and more recently their dynamic behavior. In cultured islets, interactions of beta cells with other islet cells and the matrix as well as paracrine and autocrine signaling or reaction to nutrients have been studied. Lastly, microscopy has been performed on tissue sections, visualizing the islets in an environment closer to their natural surroundings. In most efforts to date, the samples have been isolated for investigation and hence have by definition been divorced from their natural environments and deprived of vascularization and innervations. In such a setting the beta cells lack the metabolic information that is primordial to their basic function of maintaining glucose homeostasis. We review optical microscopy; its general principles, its impact in decoding beta cell function and its recent developments towards the more physiologically relevant assessment of beta cell function within the environment of the whole organism. This requires both large imaging depth and fast acquisition times. Only few methods can achieve an adequate compromise. We present extended focus Optical Coherence Microscopy (xfOCM) as a valuable alternative to both confocal microscopy and two photon microscopy (2PM), and discuss its potential in interpreting the mechanisms underlying glucose homeostasis and monitoring impaired islet function.

In vivo imaging of murine endocrine islets of Langerhans with extended-focus optical coherence microscopy.

AIMS/HYPOTHESIS: Structural and functional imaging of the islets of Langerhans and the insulin-secreting beta cells represents a significant challenge and a long-lasting objective in diabetes research. In vivo microscopy offers a valuable insight into beta cell function but has severe limitations regarding sample labelling, imaging speed and depth, and was primarily performed on isolated islets lacking native innervations and vascularisation. This article introduces extended-focus optical coherence microscopy (xfOCM) to image murine pancreatic islets in their natural environment in situ, i.e. in vivo and in a label-free condition. METHODS: Ex vivo measurements on excised pancreases were performed and validated by standard immunohistochemistry to investigate the structures that can be observed with xfOCM. The influence of streptozotocin on the signature of the islets was investigated in a second step. Finally, xfOCM was applied to make measurements of the murine pancreas in situ and in vivo. RESULTS: xfOCM circumvents the fundamental physical limit that trades lateral resolution for depth of field, and achieves fast volumetric imaging with high resolution in all three dimensions. It allows label-free visualisation of pancreatic lobules, ducts, blood vessels and individual islets of Langerhans ex vivo and in vivo, and detects streptozotocin-induced islet destruction. CONCLUSIONS/INTERPRETATION: Our results demonstrate the potential value of xfOCM in high-resolution in vivo studies to assess islet structure and function in animal models of diabetes, aiming towards its use in longitudinal studies of diabetes progression and islet transplants.

Laser Doppler imaging for intraoperative human brain mapping.

The identification and accurate location of centers of brain activity are vital both in neuro-surgery and brain research. This study aimed to provide a non-invasive, non-contact, accurate, rapid and user-friendly means of producing functional images intraoperatively. To this end a full field Laser Doppler imager was developed and integrated within the surgical microscope and perfusion images of the cortical surface were acquired during awake surgery whilst the patient performed a predetermined task. The regions of brain activity showed a clear signal (10-20% with respect to the baseline) related to the stimulation protocol which lead to intraoperative functional brain maps of strong statistical significance and which correlate well with the preoperative fMRI and intraoperative cortical electro-stimulation. These initial results achieved with a prototype device and wavelet based regressor analysis (the hemodynamic response function being derived from MRI applications) demonstrate the feasibility of LDI as an appropriate technique for intraoperative functional brain imaging.

Image formation in fluorescence coherence-gated imaging through scattering media.

Recently, we have experimentally demonstrated a new form of cross-sectional, coherence-gated fluorescence imaging referred to as SD-FCT ('spectral-domain fluorescence coherence tomography'). Imaging in SD-FCT is accomplished by spectrally detecting self-interference of the spontaneous emission of fluorophores, thereby providing depth-resolved information on the axial positions of fluorescent probes. Here, we present a theoretical investigation of the factors affecting the detected SD-FCT signal through scattering media. An imaging equation for SD-FCT is derived that includes the effects of defocusing, numerical-aperture, and the optical properties of the medium. A comparison between the optical sectioning capabilities of SD-FCT and confocal microscopy is also presented. Our results suggest that coherence gating in fluorescence imaging may provide an improved approach for depth-resolved imaging of fluorescently labeled samples; high axial resolution (a few microns) can be achieved with low numerical apertures (NA<0.09) while maintaining a large depth of field (a few hundreds of microns) in a relatively low scattering medium (6 mean free paths), whereas moderate NA's can be used to enhance depth selectivity in more highly scattering biological samples.

Extended focus depth for Fourier domain optical coherence microscopy.

We report on a new detection scheme for Fourier domain optical coherence microscopy that exhibits high transverse resolution along an axially extended focal range. Nearly constant transverse resolution of approximately 1.5 microm along a focal range of 200 microm is experimentally verified with a maximum sensitivity of 105 dB. A broad-bandwidth Ti:sapphire laser allowed for an axial resolution of 3 microm in air.

High light field confinement for fluorescent correlation spectroscopy using a solid immersion lens.

In this paper we present recent single molecule detection experiment using a solid immersion lens (SIL) for fluorescent correlation spectroscopy measurements. We compared the performance of the SIL in combination with an air objective (40x, numerical aperture (NA)=1.15) with a water immersion objective (40x, NA=0.6) in a confocal microscope system (ConfoCorr 1). Important parameters for single molecule experiments such as collection efficiency and excitation field confinement were investigated. Although the two set-ups have similar numerical aperture the measurements demonstrated higher field confinement and better collection efficiency for the SIL system in comparison to the conventional confocal set-up. Adding spherical aberrations shifts the sample volume up to 4 microm away from the plane surface of the SIL and conserves a diffraction limited focal volume. In this case the FCS autocorrelation demonstrates a free 3D diffusion of dye molecules in a highly confined light field.

Spatially incoherent illumination as a mechanism for cross-talk suppression in wide-field optical coherence tomography.

Comparison of two illumination modes for wide-field optical coherence tomography has revealed that spatially coherent illumination generates coherent cross talk, causing significant image degradation, and that spatially incoherent illumination, with an adequate interferometer design, provides an efficient mechanism for suppression of coherent cross talk. This is shown by comparison of a pulsed laser with a thermal light source for a U.S. Air Force resolution target covered with a scattering solution made from microbeads as well as for an ex vivo tooth.

Self-referenced method for optical path difference calibration in low-coherence interferometry.

A simple method for the calibration of optical path difference modulation in low-coherence interferometry is presented. Spectrally filtering a part of the detected interference signal results in a high-coherence signal that encodes the scan imperfections and permits their correction. The method is self-referenced in the sense that no secondary high-coherence light source is necessary. Using a spectrometer setup for spectral filtering allows for flexibility in both the choice of calibration wavelength and the maximum scan range. To demonstrate the method's usefulness, it is combined with a recently published digital spectral shaping technique to measure the thickness of a pellicle beam splitter with a white-light source.

High-speed femtosecond pump-probe spectroscopy with a smart pixel detector array.

A new femtosecond pump-probe spectroscopy technique is demonstrated that permits the high-speed, parallel acquisition of pump-probe measurements at multiple wavelengths. This is made possible by use of a novel, two-dimensional smart pixel detector array that performs amplitude demodulation in real time on each pixel. This detector array can not only achieve sensitivities comparable with lock-in amplification but also simultaneously performs demodulation of probe transmission signals at multiple wavelengths, thus permitting rapid time- and wavelength-resolved femtosecond pump-probe spectroscopy. Measurements on a thin sample of bulk GaAs are performed across 58 simultaneous wavelengths. Differential probe transmission changes as small as approximately 2 x 10(-4) can be measured over a 5-ps delay scan in only approximately 3 min. This technology can be applied to a wide range of pump-probe measurements in condensed matter, chemistry, and biology.

Numerical dispersion compensation for Partial Coherence Interferometry and Optical Coherence Tomography.

Dispersive samples introduce a wavelength dependent phase distortion to the probe beam. This leads to a noticeable loss of depth resolution in high resolution OCT using broadband light sources. The standard technique to avoid this consequence is to balance the dispersion of the sample byarrangingadispersive materialinthereference arm. However, the impact of dispersion is depth dependent. A corresponding depth dependent dispersion balancing technique is diffcult to implement. Here we present a numerical dispersion compensation technique for Partial Coherence Interferometry (PCI) and Optical Coherence Tomography (OCT) based on numerical correlation of the depth scan signal with a depth variant kernel. It can be used a posteriori and provides depth dependent dispersion compensation. Examples of dispersion compensated depth scan signals obtained from microscope cover glasses are presented.

Corneal morphology in vitro after superficial keratectomy with q-switched Er:YSGG and free-running Er:YAG lasers.

PURPOSE: Examination of morphology in corneal ablation induced by a q-switched Er:YSGG (2.79 microm) laser and a free-running Er:YAG laser (2.94 microm). METHODS: Defined ablation of 6-mm diameter and 15, 30, 55, 90, and 120-microm depth was performed on freshly enucleated swine eyes. Er:YSGG laser parameters: fluence 1.6 J/cm2, frequency 6 Hz, spot-size 465 microm FWHM, scanning-mode, pulse number 1520 to 6210. Er:YAG laser parameters: fluence 3 J/cm2, frequency 1.5 Hz, spotsize 6 mm, wide area ablation, pulse number 2 to 13. Corneal morphology was analyzed by gross photography, histology, scanning electron microscopy, and scanning nearfield acoustic microscopy. RESULTS: Histology showed thermal damage of 5 to 15 microm in depth caused by the Er:YSGG laser in comparison with 10 to 20 microm by the Er:YAG laser. Average roughness of the ablated surface measured with scanning nearfield acoustic microscopy was 20 to 40 microm for the Er:YSGG laser and 5 to 15 microm for the Er:YAG laser. These data confirm the subjective impression of images created by scanning electron microscopy and gross photography. CONCLUSIONS: Although the Er:YAG laser system appeared to demonstrate a smoother corneal surface than the q-switched Er:YSGG laser, the thermal damage in either case poses a potential limitation for clinical use in lamellar refractive surgery.

Actively mode-locked visible upconversion fiber laser.

Active mode locking of a Pr(3+)/Yb (3+) -doped upconversion fluoride fiber laser with an all-fiber ZnO acousto-optic phase modulator is demonstrated for the first time to the authors' knowledge. Optical pulses of ~550-ps duration with a repetition rate of 239 MHz at a wavelength of 635 nm have been generated.

[Decrease of retinal image contrast after photorefractive keratectomy, improvement within the scope of surface restitution]. / Abnahme des retinalen Bildkontrasts nach photorefraktiver Keratektomie, Besserung im Rahmen der Oberflächenrestitution.

Photorefractive keratectomy (PRK) with the ArF excimer lasers in current use usually approximates the intended corneal curvature by a mean of a delicate step-type pattern that is smooth off afterwards by reepithelialization and tear film. The present study was based on a model eye with axial myopia of -6 D but otherwise the optical and geometric properties of the Gullstrand model eye and was designed to investigate to what extent. (1) corneal step patterns can reduce retinal image contrast and (2) smoothing effects can restore such a loss. METHODS. The corneal surface resulting from PRK in the case of a myopia of -6 D (optical zone diameter 6 mm) is calculated for the parameters of the model eye. The retinal image contrasts of bar patterns are calculated by PSF (point spread function) analysis: varying size of pupil, wavelength, bar width, ablation step height and degree of smoothing. RESULTS. Step height influences retinal image contrast crucially. With step heights above 0.4 micron a massive loss of retinal image contrast must be expected, which can, however, be corrected to a useful extent by surface-smoothing effects. CONCLUSION. This study indicates that PRK with excimer lasers should be performed with low fluence and correspondingly low corneal step heights.