Statistically quantified measurement of an Alzheimer's marker by surface‐enhanced Raman scattering

Fibrillar forms of the Amyloid‐β (Aβ) protein have been implicated in the early stages of Alzheimer's disease (AD), however there are no standardised assays for soluble Aβ oligomer biomarkers that provide the best indication of the disease progression [1,2]. As a step towards a fast and label‐free method for testing different AD biomarkers, we have combined laser nano‐textured substrates with a SERS mapping technique and validated it using soluble Aβ‐40 oligomers [3‐5]. The nano‐textured SERS substrates provide fast (&5 min), label‐free spectra associated with soluble Aβ‐40 oligomers down to a concentration of 10 nM. Statistical analysis of the spectral intensities mapped over the substrate surface shows a quantitative correlation with the oligomer concentration.

Schematics of experiments: SERS mapping of Aβ‐40 (left figure: measured SERS intensity overlayed with an SEM image of ripples) was carried out on the laser nano‐textured (ripple) surface of sapphire and statistical analysis of the SERS intensity was carried out for qualitative (a high SERS intensity at low probability) and quantitative (a moderate SERS intenisty at the highest probability) measures. Quantitative statistical analysis of SERS mapping data can be performed off line for cross correlations with other known SERS signatures.     

Identification of blood species based on surface-enhanced Raman scattering spectroscopy and convolutional neural network

The identification of blood species is of great significance in many aspects such as forensic science, wildlife protection, and customs security and quarantine. Conventional Raman spectroscopy combined with chemometrics is an established method for identification of blood species. However, the Raman spectrum of trace amount of blood could hardly be obtained due to the very small cross-section of Raman scattering. In order to overcome this limitation, surface-enhanced Raman scattering (SERS) was adopted to analyze trace amount of blood. The 785 nm laser was selected as the optimal laser to acquire the SERS spectra, and the blood SERS spectra of 19 species were measured. The convolutional neural network (CNN) was used to distinguish the blood of 19 species including human. The recognition accuracy of the blood species was obtained with 98.79%. Our study provides an effective and reliable method for identification and classification of trace amount of blood.     

Bacteriochlorin‐bis(spermine) conjugate affords an effective photodynamic action to eradicate microorganisms

A novel bacteriochlorin bearing two spermine units ( BCS ) was synthesized from 3,13‐dibromo‐8,8,18,18‐tetramethylbacteriochlorin ( BC‐Br ^3,13 ). The synthesis involved the Suzuki coupling of BC‐Br ^3,13 to obtain a bacteriochlorin‐dibenzaldehyde ( BCA ), which was subjected to reductive amination with spermine. The resulting bacteriochlorin BCS presents a strong near‐infrared absorption band at 747 nm, emits at 750 nm with fluorescence quantum yield of 0.14, and generates singlet molecular oxygen, O₂(¹Δ_g), with a quantum yield of 0.27. Photokilling capacities mediated by BCS were evaluated in microbial cells. The viability of Staphylococcus aureus decreased 7 logs when cells were incubated with 1 μM BCS and irradiated for 15 minutes. Comparable photocytotoxic effect was obtained with Escherichia coli, when cells were treated for 30 minutes with visible light. BCS was also an effective photosensitizer to inactivate Candida albicans. In addition, this bacteriochlorin was able to eradicate bacteria at short incubation times. The structure of BCS contains eight basic amino groups that, when protonated in water, increase the binding to the cell envelope. In summary, the readily accessible bacteriochlorin BCS was highly effective at low concentrations as a broad‐spectrum antimicrobial photosensitizer.     

Dependence of laser-induced optical breakdown on skin type during 1064 nm picosecond laser treatment

The current study aims to evaluate the dependence of laser-induced optical breakdown (LIOB) on skin types by using 1064 nm picosecond laser with micro-lens arrays (MLA) and diffractive optical elements (DOE). Both black and white skin tissues were examined to comparatively assess the LIOB effects in the skin in terms of laser-induced vacuolization. The black skin irradiated at 3.0 J/cm² demonstrated that MLA yielded a deeper distribution (180-400 μm) of laser-induced vacuoles with a size of 67 μm, compared to DOE (180-280 μm; 40 μm in size). However, the white skin presented that MLA created larger vacuoles (134 μm in size) in a smaller number at deeper distributions (125-700 μm) than MLA with the black skin. DOE generated no laser-induced vacuolization in the white skin. The white skin tissue with inherent higher scattering could be responsible for deeper vacuolization after the picosecond laser treatment. Further investigations are expected to determine the optimal treatment conditions for various skin types.     

SERS quantitative detection of trace human chorionic gonadotropin using a label‐free Victoria blue B as probe in the aggregated immunonanogold sol substrate

Nanogold particles (NG) were modified by anti‐rabbit antibody (RAb) against human chorionic gonadotropin to obtain an immunonanogold probe (ING). In pH 7.0 Na₂HPO₄‐citrate buffer solution containing KCl, ING probes formed large aggregates in which Victoria blue B (VBB) molecules were adsorbed on the surface and which exhibited strong surface‐enhanced Raman scattering (SERS) at a peak of 1612 cm^–1. After addition of human chorionic gonadotropin (hCG) an immune reaction with the ING probe occurred to form dispersive ING–hCG complexes with non‐SERS activity that led to a decreased SERS peak at 1612 cm^–1. The decreased SERS intensity was linear to the concentration of hCG over 2.4–73.2 ng/mL. The ING reaction was studied in detail by SERS, scanning electron microscope (SEM), resonance Rayleigh scattering (RRS), surface plasmon resonance (SPR) absorption and laser scattering techniques. SERS quenching was observed and discussed. Copyright © 2014 John Wiley & Sons, Ltd.     

Superficial photothermal laser ablation of ex vivo sheep esophagus using a cone‐shaped optical fiber tip

Superficial photothermal laser ablation (SPLA) may be useful as a therapeutic approach producing a depth of injury that is sufficient to eliminate mucosal lesion but not deep enough to induce thermal effects in deeper tissue layers. The purpose of this preliminary study is twofold: (a) to describe design steps of a fiber probe capable of delivering a tightly focused laser beam, including Monte‐Carlo‐based simulations, and (b) to complete the initial testing of the probe in a sheep esophagus model, ex vivo. The cone‐shaped (tapered) fiber tip was obtained by chemical etching of the optical fiber. A 1505 nm diode laser providing power up to 500 mW was operated in continuous wave. The successful SPLA of the sheep mucosa layer was demonstrated for various speed‐power combinations, including 300 mW laser power at a surface scanning rate of 0.5 mm/s and 450 mW laser power at a surface scanning rate of 2.0 mm/s. Upon further development, this probe may be useful for endoscopic photothermal laser ablation of the mucosa layer using relatively low laser power.     

Cover Image: Plasmodium berghei sporozoites in nonreplicative vacuole are eliminated by a PI3P‐mediated autophagy‐independent pathway (Cellular Microbiology 1/2021)

Immunofluorescence image of Plasmodium berghei sporozoites (red) in HeLa cells 1.5 hours post infection. The parasitophorous vacuole membrane (PVM) that surrounds the parasites was stained with antibodies against the PVM‐resident protein UIS4 (grey). DNA was stained with DAPI (blue). Imaging was performed on a Leica SP8 confocal microscope. For further details, readers are referred to the article by Bindschedler et al. on p. e13271 of this issue.

     

Localization of PD‐L1 on single cancer cells by iSERS microscopy with Au/Au core/satellite nanoparticles

Programmed cell death‐ligand 1 (PD‐L1) is an important predictive biomarker. The detection of PD‐L1 can be crucial for patients with advanced cancer where the use of immunotherapy is considered. Here, we demonstrate the use of immuno‐SERS microscopy (iSERS) for localizing PD‐L1 on single cancer SkBr‐3 cells. A central advantage of iSERS is that the disturbing autofluorescence from cells and tissues can be efficiently minimized by red to near‐infrared laser excitation. In this study we employed Au/Au core/satellite nanoparticles as SERS nanotags because of their remarkable signal brightness and colloidal stability upon red laser excitation. False‐color iSERS images of the positive and negative controls clearly reveal the specific localization of PD‐L1 with SERS nanotag‐labeled antibodies.      

Laser-assisted nucleic acid delivery: A systematic review

Gene therapy has become an effective treatment modality for some conditions. Laser light may augment or enhance gene therapy through photomechanical, photothermal, and photochemical. This review examined the evidence base for laser therapy to enhance nucleic acid transfection in mammalian cells. An electronic search of MEDLINE, Scopus, EMBASE, Web of Science, and Google Scholar was performed, covering all available years. The preferred reporting items for systematic reviews and meta-analyses guideline for systematic reviews was used for designing the study and analyzing the results. In total, 49 studies of laser irradiation for nucleic acid delivery were included. Key approaches were optoporation, photomechanical gene transfection, and photochemical internalization. Optoporation is better suited to cells in culture, photomechanical and photochemical approaches appear well suited to in vivo use. Additional studies explored the impact of photothermal for enhancing gene transfection. Each approach has merits and limitations. Augmenting nucleic acid delivery using laser irradiation is a promising method for improving gene therapy. Laser protocols can be non-invasive because of the penetration of desirable wavelengths of light, but it depends on various parameters such as power density, treatment duration, irradiation mode, etc. The current protocols show low efficiency, and there is a need for further work to optimize irradiation parameters.     

Sialolithiasis: Application parameters for an optimal laser therapy

In‐vitro experimental parametric studies of laser ablation using natural sialoliths and artificial stones have been performed toward an efficient laser treatment of sialolithiasis. Surface microstructure and water adsorption become critical for coupling high power pulsed Ho:YAG laser radiation (λ = 2080 nm, τ ～250 μsec), inducing ablative interactions and stone fragmentation. Results reveal a generic trend, with single pulse laser energy density threshold for sialolith ablative erosion at ～200 J cm^?2 (corresponding to intensity ～800 kW cm^?2) and fragmentation rates reaching ～1 mm/pulse at ～2400 J cm^?2. This process shows no saturation, suggesting that very high energy density irradiation at low pulse repetition rate is an efficient approach. Such operation facilitates rapid cooling and minimal thermal loading of the oral and maxillofacial area, thus causing negligible adverse effects. The method is expected to contribute to the establishment of an easy and optimal therapeutic protocol for sialolithiasis pathology.     

Nanoplasmonic sensor for foodborne pathogens detection. Towards development of ISO‐SERS methodology for taxonomic affiliation of Campylobacter spp.

According to EU summary report on zoonoses, zoonotic agents and food‐borne outbreaks in 2017, Campylobacter was the most commonly reported gastrointestinal bacterial pathogen in humans in the EU. Unfortunately, the standard methods for the detection of thermotolerant Campylobacter spp. in foods are time‐consuming. Additionally, the qualified staff is obligatory. For this reason, new methods of pathogens detection are needed. The present work demonstrates that surface‐enhanced Raman scattering (SERS) is a reliable and fast method for detection of Campylobacter spp. in food samples. The proposed method combines the SERS measurements performed on an Ag/Si substrate with two initial steps of the ISO standard procedure. Finally, the principal component analysis (PCA) allows for statistical classification of the studied bacteria. By applying the proposed ISO‐SERS‐PCA method in the case of Campylobacter bacteria the total detection time may be reduced from 7 to 8 days required by ISO method to 3 to 4 days in the case of SERS‐based approach.     

Therapeutic prognosis of prostate cancer using surface-enhanced Raman scattering of patient urine and multivariate statistical analysis

Surface-enhanced Raman scattering (SERS) is highly sensitive and label-free analytical technique based on Raman spectroscopy aided by field-multiplying plasmonic nanostructures. We report the use of SERS measurements of patient urine in conjunction with biostatistical algorithms to assess the treatment response of prostate cancer (PCa) in 12 recurrent (Re) and 63 nonrecurrent (NRe) patient cohorts. Multiple Raman spectra are collected from each urine sample using monodisperse silver nanoparticles (AgNPs) for Raman signal enhancement. Genetic algorithms-partial least squares-linear discriminant analysis (GA-PLS-LDA) was employed to analyze the Raman spectra. Comprehensive GA-PLS-LDA analyses of these Raman spectral features (p = 3.50 × 10⁻¹⁶ ) yield an accuracy of 86.6%, sensitivity of 86.0%, and specificity 87.1% in differentiating the Re and NRe cohorts. Our study suggests that SERS combined with multivariate GA-PLS-LDA algorithm can potentially be used to detect and monitor the risk of PCa relapse and to aid with decision-making for optimal intermediate secondary therapy to recurred patients.     

Issue Information

Cover Image Telipogon peruvianus is a highly restricted Neotropical orchid species, endemic to the southern Peruvian Andes. It is only known from two localities, which are affected by anthropogenic disturbance. The present study investigated whether the restricted distribution of T. peruvianus has led to low genetic diversity and inbreeding, thus threatening its survival. https://doi.org/10.1111/1442-1984.12291

     

Rational design of thiolated polyenes as trifunctional Raman reporter molecules in surface‐enhanced Raman scattering nanotags for cytokine detection in a lateral flow assay

The characteristic vibrational spectroscopic fingerprint of Raman reporter molecules adsorbed on noble metal nanoparticles is employed for the identification of target proteins by the corresponding surface‐enhanced Raman scattering (SERS) nanotag‐labeled antibodies. Here, we present the modular synthesis of thiolated polyenes with two to five C═C double bonds introduced via stepwise Wittig reactions. The experimental characterization of their electronic and vibrational properties is complemented by density functional theory calculations. Highly SERS‐active nanotags are generated by using the thiolated polyenes as Raman reporter molecules in Au/Au core/satellite supraparticles with multiple hot spots. The cytokines IL‐1β and IFN‐γ are detected in a duplex SERS‐based lateral flow assay on a nitrocellulose test strip by Raman microscopy. The thiolated polyenes are suitable for use in immuno‐SERS applications such as point‐of‐care testing as well as cellular and tissue imaging.     

A quantitative evaluation of the use of medical lasers in German hospitals

The laser has become an integral part of modern medicine, procedures based on this technique have found their way into a multitude of medical disciplines. There is, however, no data available on the detailed quantitative development of laser use in the medical sector. This fact gave rise to the idea of the present study, which analyzed the raw data of the quality report of German hospitals with respect to this subject. Over the 9 years of report, a steady increase in the cumulative number of cases was evident, although not all body regions in which the medical laser is used followed this trend. The CO₂ laser was found to be the most commonly applied laser, even though a large spectrum of different laser types is used. Based on the present study, the importance of the laser for medical purposes can be confirmed.     

Biophotonic technologies for assessment of breast tumor surgical margins—A review

Breast conserving surgery (BCS) offering similar surgical outcomes as mastectomy while retaining breast cosmesis is becoming increasingly popular for the management of early stage breast cancers. However, its association with reoperation rates of 20% to 40% following incomplete tumor removal warrants the need for a fast and accurate intraoperative surgical margin assessment tool that offers cellular, structural and molecular information of the whole specimen surface to a clinically relevant depth. Biophotonic technologies are evolving to qualify as such an intraoperative tool for clinical assessment of breast cancer surgical margins at the microscopic and macroscopic scale. Herein, we review the current research in the application of biophotonic technologies such as photoacoustic imaging, Raman spectroscopy, multimodal multiphoton imaging, diffuse optical imaging and fluorescence imaging using medically approved dyes for breast cancer detection and/or tumor subtype differentiation toward intraoperative assessment of surgical margins in BCS specimens, and possible challenges in their route to clinical translation.     

Inside Cover

Transcranial photobiomodulation (tPBM) with wavelength at 808 nm was applied to a rat model of acute seizures. The tPBM successfully attenuated convulsive status epilepticus, and the tPBM was shown to reduce the apoptosis of parvalbumin‐positive interneurons (PV‐INs) and preserved the integrity of perisomatic inhibitory network of PV‐INs to principle cells in the hippocampus. Further details can be found in the article by Chung‐Min Tsai, Shwu‐Fen Chang, and Hsi Chang ( e202000095 ).

     

Accelerated inactivation of M13 bacteriophage using millijoule femtosecond lasers

Irradiation of femtosecond (fs) pulse lasers in the visible and near‐infrared ranges have been proposed as a promising approach for inactivating viruses. However, in order to achieve significant virus inactivation, past works have required relatively long irradiation times (1 hour or longer), even for small volumes. Given its advantages compared with other techniques, there is an urgent need to shorten the time required to inactivate viruses using fs laser technology. In this study, we investigate the inactivation of purified M13 bacteriophage in phosphate‐buffered saline with large active volume (1 cm³), and short exposure time (several minutes), using lasers with 20 mJ/pulse energy at various wavelengths (800, 400 nm or both 800 and 400 nm combined). For an exposure time of 15 and 2 minute, the use of a 400 nm wavelength laser results in a high load reduction of 5.8 ± 0.3 and 2.9 ± 0.15, respectively, on the log₁₀ scale of viability. We show that virus inactivation using the 400 nm laser is much more efficient compared with that using an 800 nm laser, or the simultaneous irradiation of 400 and 800 nm lasers. Higher pathogen inactivation is observed for lasers with shorter pulse duration, whereas at longer pulse durations, the inactivation is reduced. For millijoule‐energy fs laser irradiation, the M13 bacteriophage inactivation, via the reduction of the functionality of M13 bacteriophages, is accompanied with relatively small amounts of genetic damage.     

Inside Cover: Localization of PD‐L1 on single cancer cells by iSERS microscopy with Au/Au core/satellite nanoparticles (J. Biophotonics 3/2020)

iSERS (SERS=surface‐enhanced Raman scattering) microscopy is an emerging Raman‐based staining technique for the selective localization of target proteins on cells and tissues using antibody‐ SERS nanotag conjugates. In this contribution we demonstrate the feasibility of iSERS for imaging of programmed cell death‐ligand 1 (PD‐L1), an important predictive biomarker, on single SkBr‐3 breast cancer cells. Further details can be found in the article by Elzbieta Stepula, Matthias König, Xin‐Ping Wang, et al. ( e201960034 ).

     

Issue Information

ON THE COVER: Overlooked by Andean volcanos, the Bofedeles de Parinacota is an ecologically important wetland in the Chilean altiplano, where Nostoc sheets bearing diverse epimicrobiota occur in pools surrounded by bird flocks and grazing herds of camelids such as guanaco (Lama guanaco), llama (Lama glama), vicuna (Vicugna vicugna), and alpaca (Vicugna pacos).