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Simple Summary Neoplasms of the cervix are the most common types of oncological pathology. Photodynamic therapy with intravenous administration of the photosensitizer chlorin e6 shows high efficiency in the treatment of precancerous lesions of the cervix with complete eradication of the human papillomavirus. The treatment method can reduce deaths from cervical cancer and preserve fertility in patients. Spectral and video fluorescence diagnostics allows intraoperatively assessing the degree of photosensitizer accumulation and photobleaching and visualizing the boundaries of pathologically altered tissues. Abstract (1) Purpose: Improving the treatment effectiveness of intraepithelial neoplasia of the cervix associated with human papillomavirus infection, based on the application of the method of photodynamic therapy with simultaneous laser excitation of fluorescence to clarify the boundaries of cervical neoplasms. (2) Methods: Examination and treatment of 52 patients aged 22 to 53 years with morphologically and cytologically confirmed mild to severe intraepithelial cervix neoplasia, preinvasive, micro-invasive, and squamous cell cervix carcinoma. All patients were carriers of human papillomavirus infection. The patients underwent photodynamic therapy with simultaneous laser excitation of fluorescence. The combined use of video and spectral fluorescence diagnostics for cervical neoplasms made it possible to control the photodynamic therapy process at all stages of the procedure. Evaluation of the photodynamic therapy of intraepithelial cervical neoplasms was carried out with colposcopic examination, cytological conclusion, and morphological verification of the biopsy material after the photodynamic therapy course. The success of human papillomavirus therapy was assessed based on the results of the polymerase chain reaction. (3) Results. The possibility of simultaneous spectral fluorescence diagnostics and photodynamic therapy using a laser source with a wavelength of 660 nm has been established, making it possible to assess the fluorescence index in real-time and control the photobleaching of photosensitizers in the irradiated area. The treatment of all 52 patients was successful after the first photodynamic therapy procedure. According to the PCR test of the discharge from the cervical canal, the previously identified HPV types were not observed in 48 patients. Previously identified HPV types were absent after repeated PDT in four patients (CIN III (n = 2), CIS (n = 2)). In 80.8% of patients, regression of the lesion was noted. (4) Conclusions. The high efficiency of photodynamic therapy with intravenous photosensitizer administration of chlorin e6 has been demonstrated both in relation to eradication therapy of human papillomavirus and in relation to the treatment of intraepithelial lesions of the cervix.

High-grade gliomas have a diffuse and infiltrative nature of the growth of tumor cells, due to which the achievement of radical resection is difficult. Surgical resection completeness of brain tumors is an important factor in prolonging the life of patients. An accurate definition of tumor boundaries and residual fluorescent regions is impossible due to imperfections of the equipment used for fluorescent imaging. 5-aminolevulinic acid (5-ALA) is a precursor of protoporphyrin IX (PpIX) in humans and is clinically used to detect and treat tumors. Currently, fluorescence-guided surgery with PpIX used a surgical microscope with an excitation wavelength in the blue spectrum range. Because of its low ability to penetrate into biological tissue, blue light is ineffective for providing high-quality fluorescent navigation. Also, when performing an operation using radiation in the blue spectrum range, the photosensitizer’s surface layer (PS) often bleaches out, which leads to frequent errors. The use of red light emission makes it possible to slow down the PS bleaches out due to the absorption properties of PpIX, but this task is technically more complicated and requires highly sensitive cameras and specialized optical filters. The new two-channel video system for fluorescent navigation has a radiation source in the red range of the spectrum, the penetration depth of which is greater than the blue light, which makes it possible to increase the depth of probing into biological tissues. The study’s clinical part involved 5 patients with high grade glioma and 1 patient with low grade glioma: grade III oligodendrogliomas (2), grade IV glioblastomas (3), and grade II diffusion astrocytoma (1).

International audience; Background: The study proposes to improve bladder cancer diagnosis by (i) photodynamic diagnosis (PDD) using red-light excitation (632.8 nm) of 5-ALA induced-protoporphyrin IX in 9 patients’ bladder associated to the analysis of two types of signals to improve the diagnostic accuracy, and (ii) numerical modeling of scattering coefficient for providing biological explanations of the results obtained.Methods: Two modalities of the bladder cancer spectral diagnosis are presented: conventional PDD and intensity assessment of the diffusely reflected laser light by fiber-optic spectroscopy. Experiments are done in clinical conditions and as a series of numerical simulations.Results: High-grade cancerous bladder tissues display twice a higher relative fluorescence intensity (mean value 1, n = 9) than healthy (0.39, n = 9), dysplastic (0.44, n = 5) tissues and CIS (0.39, n = 2). The laser back-scattering signal allows to discriminate most effectively high-grade cancerous and dysplastic tissues from normal. Numerical modeling of diffuse reflectance spectra reveals that spectral behavior of the back-scattered light depends on both, nuclear size and nuclear density of tumoral cells.Conclusions: Unlike the fluorescence signal, where its value is higher in the case of pathological tissues, the tendency of the laser signal to, both, decrease or increase in comparison with the signal from normal urothelium, should be perceived as a sign towards the neoplasm. Numerical simulation reveals that such a double-analysis at a multiwavelength mode potentially may be used to provide diagnostic accuracy.

The new approach to intraoperative navigation during glial brain tumors removal is presented. A combined method is proposed for simultaneous spectroscopic and video fluorescence analysis of the state of tissues in the destruction zone using the applied part performed in the form of a neurosurgical aspirator cannula. In the walls of the applied part there are tubular channels into which lighting and receiving optical fibers are integrated. At the end of the cannula, the channels for optical fibers are arranged so as to perform spectroscopic analysis in contact with the surface of the biological tissue, as well as video fluorescence analysis at the working distance to the surface of the tissue. The joint use of fiber-optic systems for recording the video stream and spectral dependences allows real-time assessment of the degree of pathological tissue changes in the field of view of the video system, which are also located in the aspiration zone, with the simultaneous quantification of diagnostically significant spectroscopic criteria. System testing was carried out on samples of human intracranial tumors obtained during neurosurgical operations. During the removal of a tumor from different sites (tumor center, perifocal area), the degree of in vivo fluorescence signal from the tumor site was determined intraoperatively using a Zeiss Opmi Pentero intraoperative microscope in Blue 400 mode. From the selected area of the tumor, biopsy material was taken (presumably homogeneous in its properties) with subsequent measurement of spectra and combined images using the developed device. A high correlation was shown between the level of the fluorescence signal recorded spectroscopically and the brightness of the fluorescence image in the endoscopic channel of the device. The level of the fluorescent signal showed a high correlation with the degree of malignancy of tissues according to the results of pathomorphological examination.

We present a comprehensive analysis of the submissions to the first edition of the Endoscopy Artefact Detection challenge (EAD). Using crowd-sourcing, this initiative is a step towards understanding the limitations of existing state-of-the-art computer vision methods applied to endoscopy and promoting the development of new approaches suitable for clinical translation. Endoscopy is a routine imaging technique for the detection, diagnosis and treatment of diseases in hollow-organs; the esophagus, stomach, colon, uterus and the bladder. However the nature of these organs prevent imaged tissues to be free of imaging artefacts such as bubbles, pixel saturation, organ specularity and debris, all of which pose substantial challenges for any quantitative analysis. Consequently, the potential for improved clinical outcomes through quantitative assessment of abnormal mucosal surface observed in endoscopy videos is presently not realized accurately. The EAD challenge promotes awareness of and addresses this key bottleneck problem by investigating methods that can accurately classify, localize and segment artefacts in endoscopy frames as critical prerequisite tasks. Using a diverse curated multi-institutional, multi-modality, multi-organ dataset of video frames, the accuracy and performance of 23 algorithms were objectively ranked for artefact detection and segmentation. The ability of methods to generalize to unseen datasets was also evaluated. The best performing methods (top 15%) propose deep learning strategies to reconcile variabilities in artefact appearance with respect to size, modality, occurrence and organ type. However, no single method outperformed across all tasks. Detailed analyses reveal the shortcomings of current training strategies and highlight the need for developing new optimal metrics to accurately quantify the clinical applicability of methods.

The purpose of this study is to demonstrate the capabilities of laser spectral and video fluorescence diagnosis used for stomach tumors using 5-ALA photosensitizer. The spectroscopic method is presented with an example of a characteristic fluorescence spectrum from stomach with 5-ALA and quantitative statistics. The laser excitation wavelength was 632.8 nm. The analysis of the video system is presented with clinical statistics. The penetration depth of 3–4 mm of the He–Ne laser during the spectroscopic study allowed for scanning the mucous and submucous layers of the stomach and for detecting tumorous growths in these layers. Registration of fluorescence using the spectral system enabled surgeons to conduct express estimation of dubious stomach tissues, to make biopsy from doubtful areas to reveal precancer and early cancer states. The video fluorescence analysis with the application of 5-ALA-induced PPIX may be recommended for the use as an express method of diagnosis including early diagnosis of malignant stomach diseases as well as for intraoperative assessment of tumor extension and detection of canceromatous foci during laparoscopy. The optimal time interval for the diagnosis (regardless of the nature of the study – endoscopic, laparoscopic, or intraoperative) is 2–4 h from the administration of photosensitizer. The optimal dose of the photosensitizer is 20 mg per 1 kg of the body weight.

Novel equipment for surgery navigation thru photodynamic diagnosis is presented with laser fluorescence excitation and dual channel video registration. This equipment could be incorporated in a standard surgery tool like aspirator canule. It is based on fiberoptic endoscope with special channels for laser illumination in red and blue spectral ranges and video head based on two spectrally-resolved cameras.

International audience; A 5-ALA-induced fluorescence-based imaging device for guidance during surgery of malignant and non-malignant preliminary photosensitized tumors is presented. The setup fits existing clinical optical rigid and flexible endoscopes and operation microscopes. It consists of three light sources including white light, red light fluorescence excitation and blue light fluorescence excitation sources. The light from any combination of the latter sources is delivered to tissue using specially designed fiber optic light guide. Two cameras are used to acquire fluorescence and back reflected white light images: a gray-level camera for fluorescence in the far red range and a color camera for white light images. A dichroicmirror is implemented to spectrally split the light coming from tissue. Images from both cameras are processed into a computer with specially developed software where it can be displayed in different modes including overlaying or been used for image mosaicing which allows for increasing the intrinsic reduced field of view of endoscopes by providing highly resolved extended cartography. Experiments were carried out on phantoms and on patients in clinical conditions during surgery of brain and other tissues. Blue light excitation was more sensitive for thin tumors but red light excitation was more beneficial for solid tumors and for navigation in presence of slight bleeding.

Background: In excess of 60% of all cancers are detected in low and middle-income countries, with breast cancer (BC) the dominant malignancy for women. Incidence rates continue to climb, most noticeably in the less than 50-year-old population. Expansion of mammography infrastructure and resources is lacking, resulting in over 60% of women diagnosed with stage III/IV BC in the majority of these countries. Optical Breast Spectroscopy (OBS) was shown to correlate well with mammographic breast density (MBD). OBS could aid breast screening programs in low- and middle-income countries by lowering the number of mammographs required for complete population coverage. However, its performance needs to be tested in large population trails to ensure high sensitivity and acceptable specificity. Methods: For the planned studies in low- and middle-income countries in different continents, online methods need to be implemented to monitor the performance and data collection by these devices, operated by trained nurses. Based on existing datasets, procedures were developed to validate an individual woman's data integrity and to identify operator errors versus system malfunctions. Results: Using a dataset comprising spectra from 360 women collected by 2 instruments in different locations and with 3 different trained operators, automated methods were developed to identify 100% of the source or photodetector malfunctions as well as incorrect calibrations and 96% of instances of insufficient tissue contact. Conclusions: Implementing the dataset validation locally in each instrument and tethered to a cloud database will allow the planned clinical trials to proceed.

We evaluated diffuse reflectance spectroscopy implemented as a small field-of-view technique for discrimination of dysplasia from metaplasia in Barrett’s esophagus as an adjuvant to autofluorescence endoscopy. Using linear discriminant analysis on 2579 spectra measured in 54 patients identified an optimum a 4-wavelength classifier (at 485, 513, 598 and 629 nm). Sensitivity and specificity for a test data set were 0.67 and 0.85, respectively. Spectroscopic results show that this technique could be implemented in wide-field imaging mode to improve the accuracy of existing endoscopy techniques for finding early pre-malignant lesions in Barrett’s esophagus. Results show that the discrimination occurs likely due to redistribution of blood content in the tissue sensed by the optical probing with the wavelength-dependent sampling depth. (© 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim)

Background:Fluorescence endoscopy systems for photosensitizer visualization have proved to be powerful tools for highlighting malignant tumor boundaries as well as detecting small, visually non-detectable, residual parts during photodynamic therapy. Most of these devices use excitation wavelengths in the blue visual spectrum range (405 nm) which limits the penetration depth in the tissue.Objective:In the study being presented in this article an apparatus and a method were developed for performing endoscopic fluorescence diagnostics of photosensitizer accumulation using excitation light in the red part of visual spectrum, i.e., 635 nm, which allows not only a deeper penetration of light into the tissue but also better scanning abilities and a higher diagnostic quality. Additionally, 635-nm radiation can penetrate thin layers of blood which appear during surgery.Material and methods:In order to use 635-nm excitation, a specially designed video endoscopy system was developed. The key feature of the video system is a dual camera video receiver where one sensitive B/W camera receives the fluorescence signal and a color camera receives the real-time image in natural colors during navigation. The software developed for the apparatus allows overlaying of the video output of fluorescence image on top of the conventional color image in real-time. The experimental setup and method were tested on Intralipid-based phantoms with protoporphyrin IX (PpIX) concentrations of 0.5–5 mg/kg, and then on two patients during surgery. The patients were administered 20 mg/kg 5-ALA photosensitizer 3 h before surgery according to standard practice of 5-ALA in neurosurgery.Results:The experiments demonstrate that the designed setup is sensitive enough for clear visualization of biological concentrations of PpIX in both phantoms with 0.5 mg/kg PpIX and previously photosensitized tissues of patients.Conclusion:Further prospective validation is needed to translate the results to clinical practice.

We evaluated diffuse reflectance spectroscopy implemented as a small field-of-view technique for discrimination of dysplasia from metaplasia in Barrett's esophagus as an adjuvant to autofluorescence endoscopy. Using linear discriminant analysis on 2579 spectra measured in 54 patients identified an optimum a 4-wavelength classifier (at 485, 513, 598 and 629 nm). Sensitivity and specificity for a test data set were 0.67 and 0.85, respectively. Spectroscopic results show that this technique could be implemented in wide-field imaging mode to improve the accuracy of existing endoscopy techniques for finding early pre-malignant lesions in Barrett's esophagus. Results show that the discrimination occurs likely due to redistribution of blood content in the tissue sensed by the optical probing with the wavelength-dependent sampling depth.

Development of minimal invasive diagnostics methods is of great importance in today's healthcare. The present work deals with the measurements of glucose concentration in small drops, its volume being in the range of 1 (mu) L and its glucose concentration being in the range of 1-10 mM. The aim of the work was to develop robust quantitative method of glucose measurement in small volumes of interstitial fluid obtained after laser shot or by other low invasive procedures. The commercially available Amlex Red glucose assay kit was used in the present work. The glucose is detected by enzyme-coupling reactions resulting in formation of fluorescence dye resorufin. The diagnostic paper was used as a reaction medium. The fluorescence of dye in diagnostic paper was measured with the use of fiber optics spectrometer. It was shown that dye fluorescence correlates fairly well with glucose concentration. The method may be modified to involve cholesterol concentration measurement as well.