Front Cover

This work studied two-photon collagen crosslinking (CXL) in human corneal lenticules induced by near infrared femtosecond laser, possessing the advantages of multiphoton absorption and near-infrared light. The corneal stiffness can be enhanced by up to 296% without significantly reducing corneal transparency (<3%), paving the way to treat ophthalmic disorders such as keratoconus by 3D CXL of in vivo human cornea with higher safety, precision and efficacy. Further details can be found in the article by Zhenzhou Cheng, Nan Zhang, Le Chang, Pengfei Qi, Lin Zhang, Lie Lin, Yan Wang, and Weiwei Liu ( e202200160 ).

     

The AC conductivity of human sweat ducts as the dominant factor in the sub-THz reflection coefficient of skin

The helical nature of human sweat ducts, combined with the morphological and dielectric properties of skin, suggests electromagnetic activity in the sub-THz frequency band. A detailed electromagnetic simulation model of the skin, with embedded sweat ducts, was created. The model includes realistic dielectric properties based on the measured water content of each layer of skin, derived from Raman Spectroscopy. The model was verified by comparing it to measurements of the reflection coefficient of the palms of 13 volunteers in the frequency band 350–410 GHz. They were subjected to a measurement protocol intended to induce mental stress, thereby also activating the sweat glands. The Galvanic Skin Response was concurrently measured. Using the simulation model the optimal ac-conductivity for each measurement was found. The range of variation for all subjects was found to be from 100 S/m to a maximum value of 6000 S/m with averages of 1000 S/m. These are one order of magnitude increase from the accepted values for water at these frequencies (~100 s/m at 100 GHz). Considering the known biochemical mechanism for inducing perspiration, we conclude that these ac-conductivity levels are probably valid, even though the real time measurements of sweat ac-conductivity levels inside the duct are inaccessible.     

Lightsheet fluorescence lifetime imaging microscopy with wide‐field time‐correlated single photon counting

We report on wide‐field time‐correlated single photon counting (TCSPC)‐based fluorescence lifetime imaging microscopy (FLIM) with lightsheet illumination. A pulsed diode laser is used for excitation, and a crossed delay line anode image intensifier, effectively a single‐photon sensitive camera, is used to record the position and arrival time of the photons with picosecond time resolution, combining low illumination intensity of microwatts with wide‐field data collection. We pair this detector with the lightsheet illumination technique, and apply it to 3D FLIM imaging of dye gradients in human cancer cell spheroids, and C. elegans.     

Dual‐ and single‐shot susceptibility ratio measurements with circular polarizations in second‐harmonic generation microscopy

Polarization‐resolved second‐harmonic generation (P‐SHG) microscopy is a technique capable of characterizing nonlinear optical properties of noncentrosymmetric biomaterials by extracting the nonlinear susceptibility tensor components ratio , with z‐axis parallel and x‐axis perpendicular to the C₆ symmetry axis of molecular fiber, such as a myofibril or a collagen fiber. In this paper, we present two P‐SHG techniques based on incoming and outgoing circular polarization states for a fast extraction of : A dual‐shot configuration where the SHG circular anisotropy generated using incident right‐ and left‐handed circularly‐polarized light is measured; and a single‐shot configuration for which the SHG circular anisotropy is measured using only one incident circular polarization state. These techniques are used to extract the of myosin fibrils in the body wall muscles of Drosophila melanogaster larva. The results are in good agreement with values obtained from the double Stokes‐Mueller polarimetry. The dual‐ and single‐shot circular anisotropy measurements can be used for fast imaging that is independent of the in‐plane orientation of the sample. They can be used for imaging of contracting muscles, or for high throughput imaging of large sample areas.     

In vivo detection of tumor boundary using ultrahigh‐resolution optical coherence angiography and fluorescence imaging

Accurate detection of early tumor margin is of great preclinical and clinical implications for predicting the survival rate of subjects and assessing the response of tumor microenvironment to chemotherapy or radiation therapy. Here, we report a multimodality optical imaging study on in vivo detection of tumor boundary by analyzing neoangiogenesis of tumor microenvironment (microangiography), microcirculatory blood flow (optical Doppler tomography) and tumor proliferation (green fluorescent protein [GFP] fluorescence). Microangiography demonstrates superior sensitivity (77.7 ± 6.4%) and specificity (98.2 ± 1.7%) over other imaging technologies (eg, optical coherence tomography) for tumor margin detection. Additionally, we report longitudinal in vivo imaging of tumor progression and show that the abrupt tumor cell proliferation did not occur until local capillary density and cerebral blood flow reached their peak approximately 2 weeks after tumor implantation. The unique capability of longitudinal multimodality imaging of tumor angiogenesis may provide new insights in tumor biology and in vivo assessment of the treatment effects on anti‐angiogenesis therapy for brain cancer.     

Determination and correction of aberrations in full field optical coherence tomography using phase gradient autofocus by maximizing the likelihood function

A method for numerical estimation and correction of aberrations of the eye in fundus imaging with optical coherence tomography (OCT) is presented. Aberrations are determined statistically by using the estimate based on likelihood function maximization. The method can be considered as an extension of the phase gradient autofocusing algorithm in synthetic aperture radar imaging to 2D optical aberration correction. The efficacy of the proposed method has been demonstrated in OCT fundus imaging with 6λ aberrations. After correction, single photoreceptors were resolved. It is also shown that wave front distortions with high spatial frequencies can be determined and corrected.     

Issue Information

Cover Image An infl orescence of Clerodendrum izuinsulare (left) and C. trichotomum (right). Photos by Natsuki Aihara in Niijima Island (left) and by Takashi Miyake in Izu Peninsula (right), Japan. Miyake et al., doi.org/10.1111/1442-1984.12287

     

Issue Information

Cover Photograph: Upper left: Newt (P. waltl) juveniles and Xenopus froglets were subjected to agarose-embedded heat-shock (AeHS) method. GFP expression was induced specifically in the regenerating limbs (see Matsubara et al. 86–93). Upper right: Schematic diagram of the intermittent hypoxia exposure model. Pimonidazole-positive cells were detected in the pulmonary tissues of Xenopus laevis during repeated dives, indicating tissue hypoxia (see Fujiyama, 94–99). Lower left: The intercalary element of Zhangixalus schlegelii (arrowheads). Such skeletal elements are seen between the terminal and subterminal phalanges of all the digits of arboreal frogs (see Nakanishi, 100–108). Lower right: Tetraspanin9 expressions in Xenopus laevis (above) and X. tropicalis embryo (below) hybridized in the same tube. (see Kuriyama and Tanaka, 109–119). Bottom right corner: A 48 hpf early four-armed pluteus taken through a confocal microscope. Catecholamine-positive cells are colored in yellow (see Kalachev and Tankovich, 120–131).

     

Comparative analysis of dermal collagen and lipids in cereblon ablated mice using a multimodal nonlinear optical system

By utilizing a multimodal nonlinear optical system that combines coherent anti-Stokes Raman scattering and second harmonic generation to investigate biological characteristics of dermal tissues ex vivo, we demonstrate the potential feasibility of using this optical approach as a powerful new investigative tool for future biomedical research. For this study, our optical system was utilized for the first time to analyze lipid and collagen profiles in cereblon knockout (KO) mouse skin, and we were able to discover significant alterations in the number of carbon–carbon double bonds (wild-type vs. cereblon KO; N_CC: 0.75 vs. 0.85) of skin fatty acids in triacylglycerides as well as changes in dermal collagen fibers (25% reduction in cereblon KO). By adopting our optical system to biological studies, we provide researchers with another diagnostic approach to validate their experimental results, which will significantly advance the state of biomedical research.     

Unique spectral signature of human cutaneous squamous cell carcinoma by photoacoustic imaging

Cutaneous squamous cell carcinoma (cSCC) is a common skin cancer with metastatic potential. To reduce reoperations due to nonradical excision, there is a need to develop a technique for identification of tumor margins preoperatively. Photoacoustic (PA) imaging is a novel imaging technology that combines the strengths of laser optics and ultrasound. Our aim was to determine the spectral signature of cSCC using PA imaging and to use this signature to visualize tumor architecture and borders. Two‐dimensional PA images of 33 cSCCs and surrounding healthy skin were acquired ex vivo, using 59 excitation wavelengths from 680 to 970 nm. The spectral response of the cSCCs was compared to healthy tissue, and the difference was found to be greatest at wavelengths in the range 765 to 960 nm (P < .05). Three‐dimensional PA images were constructed from spectra obtained in the y‐z plane using a linear stepper motor moving along the x‐plane. Spectral unmixing was then performed which provided a clear three‐dimensional view of the distribution of tumor masses and their borders.     

Issue Information

Left: Isodon shikokianus and Bombus diversus Right: Isodon shikokianus and Bombus honshuensis Both Photos by Ikumi Dohzono at Mt. Hyonosen, Hyogo Prefecture, Japan. Ogishima, M., Hoshino, Y., Horie, S., Yamashiro, T., Maki, M., Suzuki, K., & Dohzono, I. (2022). Secondary contact and adaptation to local pollinator assemblages mediate geographical variation in corolla length in Isodon shikokianus. Plant Species Biology, 222–230. https://doi.org/10.1111/1442-1984.12370

     

Respirometer in a box: development and use of a portable field respirometer for estimating oxygen consumption of large-bodied fishes

This study developed a portable, low-cost field respirometer for measuring oxygen consumption rates of large-bodied fishes. The respirometer performed well in laboratory tests and was used to measure the oxygen consumption rates ( O₂) of bull sharks Carcharhinus leucas (mean: 249.21 ± 58.10 mg O₂ kg⁻¹ h⁻¹ at 27.05°C). Interspecific comparisons and assessments of oxygen degradation curves indicated that the respirometer provided reliable measurements of O₂. This system presents a field-based alternative to laboratory respirometers, opening opportunities for studies on species in remote localities, increasing the ability to validate physiological field studies.     

Issue Information

Cover Image The pistillate-phase flowers of Artabotrys (Annonaceae): A. blumei (left) and A. brachypetalus (right). Photos by Junhao Chen, taken in Hong Kong and South Africa, respectively (DOI: 10.1111/1442-1984.12273 ).

     

Anatomie und biologie der augenlosen landlungenschnecke caecilioides acicula müll

Zusammenfassung Die völlig pigmentlose Caecilioides acicula/ besitzt eine außerordentlich schmale, dreigeteilte Sohle mit sehr breitem Mittelfeld. Der Hautmuskelschlauch tritt gegenüber den großen Bluträumen des Fußes stark zurück. Für den Columellariskomplex ist die Länge der einzelnen Muskelbänder bemerkenswert. Die Lokomotionsgeschwindigkeit der Caecilioides acicula übertrifft diejenige aller anderen daraufhin untersuchten Landschnecken ganz bedeutend. Der Darmtraktus ist dem der Clausilien ähnlich. Am Dünndarm findet sich ein hier erstmalig nachgewiesenes, winziges Divertikel, dessen Funktion unklar ist. An der schlauchartigen Leber fällt die geringe Entwicklung der inneren Oberfläche auf. Der obere, apikal gelegene Leberlappen ist bedeutend größer wie der untere. Am Grunde der langgestreckten Lungenhöhle liegt die eigenartig gestaltete Niere. Der geschlossene, sekundäre Ureter mündet in zwei kurzen Asten in die Kloakenhöhle bzw. in den Atemgang. Die große Kloakenhöhle führt von Zeit zu Zeit rhythmische Verschlußbewegungen aus. Ihr liegt außen ein großer Drüsenkomplex auf, die Kloakendrüse. Die Ganglien des Schlundringes sind wenig konzentriert. Ein in die Unterseite des Pharynx eintretendes Nervenpaar zeigt gangliöse Anschwellungen, wie she his jetzt nur bei wenigen Stylommatophoren beobachtet worden sind. Tast-, Geruch- und Wärmesinn sind gut entwickelt und wichtig zur Orientierung im Raum. Die Reaktion auf optische Reize ist nicht ganz so deutlich ausgeprägt. Die Ausführwege des Genitalapparates sind stark drüsig. Die Kopulationswerkzeuge entbehren jeglicher Anhangsorgane. In bezug auf Größe und Zahl der Eier verhält sich Caecilioides acicula wie ein echtes Höhlentier. Auch die durch subterrane Lebensweise bedingte Pigment-und Augenlosigkeit hat these Schnecke mit vielen Troglobien gemeinsam. Caecilioides acicula ist eine Ferussaciide. Thieles Einreihung der Ferussaciidae unter die Achatinacea ist nach dem gegenwärtigen Stand unserer Kenntnis berechtigt.Zeiehenerklärung. a Ausführagänge der Fußdrüsenzellen - af After - ant ant Äste des Nervus tentacularis - ao Aorta - ao Außenseite der Lungendecke - at Atrium - atg ausgestülpte Genitalatrien - bc Buccalkommissur - bf bogenförmige Furche - bg Buccalganglien - bh Bindegewebshülle - bk Bindegewebskerne - blu Bluträume - bz Blutraum über der Zwitterdrüse - c Cuticula - ca Anbeftung der Spindelmuskulatur - cc Cerebralkommissur - cg Cerebralganglien - cl cr linkes, bzw. rechtes Band des Spindelmuskels - co Spindehnuskulatur - cpc Cerebropedalkonnektiv - cpl Cerebropleuralkonnektiv - d Dünndarm - de Eiweißdrüsen der Haut - di Dünndarmdivertikel - dk Kalkdrüsen - dph Diaphragma - e Wandepithel der Statozyste - ed Enddarm - ei Ei im Uterus - eiz Eizelle - eiw Eiweißmasse - ep Wandepithel des Ovidukts - es Eischale - ew Eiweißdrüse - fd Fußdrüse - fdd dorsale Drüsenzellen der Fußrüse - fdm Mündung der Fußdrüse - fl Flimmerbesatz - fr Fußsaumrinne - fv ventrale Drüsenzellen der Fußdrüse - g Geschlechtsöffnung - in innerer Nierenporus - k Kiefer - kd Kloakendrüse - kl Kloake - kl ₁ äußere Öffnung der Kloake - kld Drüsenzellen der Kloakendrüse - klh Kloakenhöhle - kv Atrioventricular-Klappe - ldo Divertikel des oberen Leberlappens - ldu Divertikel des unteren Leberlappens - lh Lungenhöhle - llo oberer Leberlappen - llu unterer Leberlappen - lo obere Lebermündung - lu untere Lebermündung - lpar linkes Parietalganglion - lpl linkes Pleuralganglion - lppac linkes Pleuroparietal-Konnektiv - lr untere Längsrinne - m Mundöffnung - ma Magen - ml Mundlappen - mm Retractor tentaculi minoris - mp Retractor pedis - mph Pharynxretraktor - mr Mantelrinne - ms Magenblindsack - mrl Lippenretraktoren - mt Retractor tentaculi majoris - mw Mantelwulst - nr Nackenrinnen - ns Nierensack - ns ₁ innerer Teil des Nierensackes - nst Nervus staticus - nt Nervus tentacularis - oe Oesophagus - ov Ovidukt - pah hintere Papille des Penisschlauches - pav vordere Papille des Penisscalauches - pc hintere Pedalkommissur - pd ₁ distales Drüsenpolster der Mantelrinne - pd ₂ proximales Drüseupolster der Mantelrinne - pe Penis - per Pericard - per unter dem Nierensack gelegener Teil des Pericards - pg Pedalganglien - ph Pharynx - plp Pleuropedalkonnektiv - pn Atemloch - po Periostracum - pro Prostata - prom Einmündung eines längsgetroffenen Prostataschlauches in die Samenrinne - pvc Parietovisceral-Konnektiv - rc Receptaculum seminis - rg Renopericardialgang - rk Riesenkerne - rkg Kern einer Riesenzelle des Pedalganglions - rp Penisretraktor - rppac rechtes Pleuroparietal-Konnektiv - rpar rechtes Parietalganglion - rpl rechtes Pleuralganglion - rs Sohlenrinne - s Seitenfeld der Fußsohle - sb Stützbalken der Radula - sd Soblendrüsen - sf Fermentzellen der Speicheldrüsen - sg Speichelgang - sh hintere Muskelsepten des Penis - sk Sekretmassen - skg gelb-grimes Sekret - sl Schleim der Fußdrüse - sld Schleimdrüsen der Haut und des Mantelrandes - slz Schleimzelle der Speicheldrüse - sor Sohlenrinne - spd Speicheldrüsen - spo Spermovidukt - sr Samenrinne - sta Statozyste - sv vordere Muskelsepten des, Penis - syk Syncytialkerne - t unterer Tentakel - tg Ganglion des Nervus tentacularis - ud Uterusdrüsen - uro oberer Ausführgang des sekundären Ureters - urp primärer Ureter - urs sekundärer Ureter - uru unterer Ausführgang des sekundären Ureters - v Ventrikel - v2, v3, v4 Nerven des Visceralgangglions - vag Vagina - vd Vas deferens - vi Visceralganglion - vml vorderer Mantellappen - vp Vena pulmonalis - zg Zwittergang - zgu aufgeknäuelter, unterer Teil des Zwitterganges - zw Zwitterdrüse     

Stereotaxic endoscopy for the ocular imaging of awake,freely moving animal models

Stereotaxic instruments are increasingly used in research animals for the study of disease, but typically require restraints and anesthetic procedures. A stereotaxic head mount that enables imaging of the anterior chamber of the eye in alert and freely mobile mice is presented in this study. The head mount is fitted based on computed tomography scans and manufactured using 3D printing. The system is placed noninvasively using temporal mount bars and a snout mount, without breaking the skin or risking suffocation, while an instrument channel stabilizes the ocular probes. With a flexible micro‐endoscopic probe and a confocal scanning laser microscopy system, <20 μm resolution is achieved in vivo with a field of view of nearly 1 mm. Discomfort is minimal, and further adaptations for minimally invasive neuroscience, optogenetics and auditory studies are possible.     

ISOLATION OF HYDROPHOBIC PROTEINS BINDING AMINO ACIDS: γ-AMINOBUTYRIC ACID BINDING IN THE RAT CEREBRAL CORTEX

—The binding of [¹⁴C]GABA to nerve-ending membranes isolated from rat cerebral cortex follows a hyperbolic curve saturating at 0·4pmol/μg protein. This binding is about 60% inhibited by chloropromazine, and about 40%, inhibited by bicuculline. A hydrophobic protein fraction binding [¹⁴C]GABA was separated from the total. lipid extract of nerve-ending membranes. The binding follows a hyperbolic curve that saturates at 10·5 pmol of [¹⁴C]GABA/μg of protein, with an apparent K_d= 30 μm . The binding is competitively inhibited by bicuculline with a K_i= 273 μm . These results are compared with those previously obtained on a GABA binding protein from crustacean muscle.     

Die V?gel B?hmens

Ohne ZusammenfassungHerr Dr. Fritsch, Verfasser des kürzlich vollendeten bekannten Werkes die Vögel Europas, veröffentlicht gegenwärtig, gestützt auf langjährige Erfahrungen, in dem Archiv für die naturwissenschaftliche Landesdurchforschung von Röhmen eine schätzenswerthe Zusammenstellung der Wirbelthiere Böhmens. Das Folgende begreift die Abtheilung der Vögel. Indem wir dieselbe dem ornithologischen Publikum zugänglich zu machen wünschen, verweisen wir für die Synonymie und das Weitere in Bezug der einzelnen Arten zugleich gern auf das oben augeführte ausführliche Werk des Verfassers.     

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.     

Front Cover: Photoacoustic imaging of the human placental vasculature (J. Biophotonics 4/2020)

Successful therapy of twin‐to‐twin transfusion syndrome requires accurate imaging to guide laser photocoagulation of the anastomosing placental vessels. Photoacoustic (PA) imaging is an alternative imaging method that provides contrast for hemoglobin, and in this study, it was used to visualize chorionic superficial and subsurface vasculature in human placentas. The strong potential of PA imaging to guide minimally invasive fetal therapies was demonstrated. Further details can be found in the article by Efthymios Maneas, Rosalind Aughwane, Nam Huynh, et al. ( e201900167 ).

     

Gold coated contact lens-type ocular in vivo dosimeter (CLOD) for monitoring of low dose in computed tomography: A Monte Carlo study

PurposeThis study reports a sensitivity enhancement of gold-coated contact lens-type ocular in vivo dosimeters (CLODs) for low-dose measurements in computed tomography (CT).MethodsMonte Carlo (MC) simulations were conducted to evaluate the dose enhancement from the gold (Au) layers on the CLODs. The human eye and CLODs were modeled, and the X-ray tube voltages were defined as 80, 120, and 140 kVp. The thickness of the Au layer attached to a CLOD ranged from 100 nm to 10 μm. The thickness of the active layer ranged from 20 to 140 μm. The dose ratio between the active layer of the Au-coated CLOD and a CLOD without a layer, i.e., the dose enhancement factor (DEF), was calculated.ResultsThe DEFs of the first 20-μm thick active layer of the 5-μm thick Au-coated CLOD were 18.4, 19.7, 20.2 at 80, 120, and 140 kVp, respectively. The DEFs decreased as the thickness of the active layer increased. The DEFs of 100-nm to 5-μm thick Au layers increased from 1.7 to 5.4 for 120-kVp X-ray tube voltage when the thickness of the active layer was 140 μm.ConclusionsThe MC results presented a higher sensitivity of Au-coated CLODs (∼20-times higher than that of CLODs without a gold layer). Au-coated CLODs can be applied to an evaluation of very low doses (a few cGy) delivered to patients during CT imaging.