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Maximal safe resection is the standard of care in the neurosurgical treatment of high-grade gliomas. To aid surgeons in the operating room, adjuvant techniques and technologies centered around improving intraoperative visualization of tumor tissue have been developed. In this review, we will discuss the most advanced technologies, specifically fluorescence-guided surgery, intraoperative imaging, neuromonitoring modalities, and microscopic imaging techniques. The goal of these technologies is to improve detection of tumor tissue beyond what conventional microsurgery has permitted. We describe the various advances, the current state of the literature that have tested the utility of the different adjuvants in clinical practice, and future directions for improving intraoperative technologies.

We developed an imaging method utilizing the available RIs. We developed two types of real-time RI imaging systems (RRIS), one for macroscopic imaging and the other for microscopic imaging. The principle of visualization was the same, converting the radiation to light by a Cs(Tl)I scintillator deposited on a fiber optic plate (FOS). Many nuclides were employed, including 14C, 18F, 22Na, 28Mg, 32P 33P, 35S, 42K, 45Ca, 48V, 54Mn, 55Fe, 59Fe, 65Zn, 86Rb, 109Cd, and 137Cs.Since radiation can penetrate the soil as well as water, the difference between soil culture and water culture was visualized. 137Cs was hardly absorbed by rice roots growing in soil, whereas water culture showed high absorption, which could provide some reassurance after the Fukushima Nuclear Accident and could indicate an important role of soil in firmly adsorbing the radioactive cesium.28Mg and 42K, whose production methods were presented, were applied for RRIS to visualize the absorption image from the roots. In addition to 28Mg and 42K, many nuclides were applied to image absorption in the roots. Each element showed a specific absorption speed and accumulation pattern. The image analysis of the absorption of Mg is presented as an example. Through successive images of the element absorption, phloem flow in the aboveground part of the plant was analyzed. The element absorption was visualized not only in the roots but also in the leaves, a basic study of foliar fertilization.In the case of the microscopic imaging system, a fluorescence microscope was modified to acquire three images at the same time: a light image, fluorescent image, and radiation image. Although the resolution of the image was estimated to be approximately 50 μm, superposition showed the expression site of the transporter gene and the actual 32P-phosphate absorption site to be the same in Arabidopsis roots.

Living cells dynamically change their morphology and function according to the cell cycle. Long-term observations of living cells are privileged when we spy the unique, cell cycle-driven molecular events, which cannot be obtained from short-term ones. Mg2+, a metal ion abundant in cells, has been shown to be involved in a variety of physiological phenomena by noninvasive cellular observation using fluorescence microscopy. However, long-term observation of Mg2+ in cells has been a great challenge. Herein, we present a protocol for the long-term microscopic imaging of intracellular Mg2+ levels using a small molecule-protein hybrid fluorescent probe we developed.

We present an innovative metalens array design with a controllable angle of view. Our design leverages the angular dependent transmission of the metastructure, and enables a compact system for large field-of-view microscopic imaging.

Safely maximizing extent of resection has become the central goal in glioma surgery. Especially in eloquent cortex, the goal of maximal resection is balanced with neurological risk. As new technologies emerge in the field of neurosurgery, the standards for maximal safe resection have been elevated. Fluorescence-guided surgery, intraoperative magnetic resonance imaging, and microscopic imaging methods are among the most well-validated tools available to enhance the level of accuracy and safety in glioma surgery. Each technology uses a different characteristic of glioma tissue to identify and differentiate tumor tissue from normal brain and is most effective in the context of anatomic, connectomic, and neurophysiologic context. While each tool is able to enhance resection, multiple modalities are often used in conjunction to achieve maximal safe resection. This paper reviews the mechanism and utility of the major adjuncts available for use in glioma surgery, especially in tumors within eloquent areas, and puts forth the foundation for a unified approach to how leverage currently available technology to ensure maximal safe resection.

International audience; Precise spatial characterization of vectorial beams is crucial for many advanced optical experiments, but challenging when wavefront and polarization features are involved together. Here we propose a reference-free method aimed at extracting the map of the complexamplitude components of any coherent beam at an optical-microscopy resolution. Our method exploits recent advances in ptychographic imaging approaches. We emphasize its versatility by reconstructing successfully various experimental vectorial beams including polarization and phase vortices, the exit field of a multicore fiber and a speckle pattern.

Breast diseases with many distinct histopathological types are showing a rising trend in incidence for decades worldwide. The proliferation of cells and the remodeling of collagen fibers in breast carcinoma tissues may be used to predict breast disease diagnosis, prognosis of treatment, and patient survival. Pathologists can label related typical pathological features as cell nuclei, aligned collagen, and disorganized collagen in hematoxylin and eosin (HE) sections of breast tissues. In this study, we apply the Mueller matrix microscopic imaging to various breast pathological section samples, and calculate corresponding polarimetry basis parameters (PBPs). A pixel-based extraction approach of polarimetry feature parameters (PFPs) is proposed using a mutual information (MI) method and a linear discriminant analysis (LDA) classifier. The three PFPs derived by the proposed learning algorithm are the simplified linear combinations of PBPs with physical meanings, and provide quantitative characterization of the three pathological features in different breast tissues respectively. We present results of the three PFPs of tissue samples from a cohort of 32 clinical patients diagnosed as normal, breast fibroma, breast ductal carcinoma in situ, invasive ductal carcinoma, and breast mucinous carcinoma with analysis of 210 regions-of-interest (ROI). The results demonstrate that the three PFPs of each breast disease tissue have specific value ranges, which has a potential to quantitatively distinguish typical pathological features between different breast tissues. This technique has good prospects for automation of the microstructure identification and prediction of breast disease diagnosis, resulting in the reduction of pathologists’ workload.

Performing long-term cell observations is a non-trivial task for conventional optical microscopy, since it is usually not compatible with environments of an incubator and its temperature and humidity requirements. Lensless holographic microscopy, being entirely based on semiconductor chips without lenses and without any moving parts, has proven to be a very interesting alternative to conventional microscopy. Here, we report on the integration of a computational parfocal feature, which operates based on wave propagation distribution analysis, to perform a fast autofocusing process. This unique non-mechanical focusing approach was implemented to keep the imaged object staying in-focus during continuous long-term and real-time recordings. A light-emitting diode (LED) combined with pinhole setup was used to realize a point light source, leading to a resolution down to 2.76 μm. Our approach delivers not only in-focus sharp images of dynamic cells, but also three-dimensional (3D) information on their (x, y, z)-positions. System reliability tests were conducted inside a sealed incubator to monitor cultures of three different biological living cells (i.e., MIN6, neuroblastoma (SH-SY5Y), and Prorocentrum minimum). Altogether, this autofocusing framework enables new opportunities for highly integrated microscopic imaging and dynamic tracking of moving objects in harsh environments with large sample areas.

Electrohydrodynamic printing (EHDP) is able to precisely manipulate the position, size, and morphology of micro/nanofibers, and fabricate high resolution scaffolds using viscous biopolymer solutions. In order to ensure the reliable printing, it is very necessary to identify EHDP cone status during fabrication. In this work, we used a digital microscopic imaging technique to monitor EHDP cones during printing, with subsequent image processing. A convolutional neural network (CNN) is then developed to classify these EHDP cones. With the help of this monitoring and identification system, scaffolds with diverse triangular, hexagonal and coil-wall microstructures can be fabricated for tissue engineering applications.

As an important biomarker for early diagnosis of cancers, sensitive detection and high-resolution imaging of microRNA-21 in cancer cells have become important and challengeable. In this work, highly sensitive detection and spatial imaging of intracellular microRNA-21 were realized by the reduced signal background through vertical polarization excitation with a polarizer. The lateral local surface plasmon resonance property of gold nanorods (AuNRs) displayed a pronounced green color with low scattering intensity, which was adjusted to red color with strong scattering intensity when the core-satellite gold nanoparticle (AuNP) assembly was constructed on the side of AuNRs through a catalyzed hairpin assembly (CHA) circuit in the presence of microRNA-21. This unique approach allows for effectively reducing the strong background signal to improve the sensitivity of detection. Additionally, the proposed strategy can not only realize the sensitive detection of microRNA-21 with the limit of detection as low as 2 pM (3σ) but also achieve the high spatial imaging of cancer cells, which provided a specific strategy for the construction and imaging of intracellular imaging probes. It is believed that the simple and sensitive approach on the basis of lateral local surface plasmon resonance property of anisotropic AuNRs with excellent sensitivity combined with high spatial imaging holds promising potentials to visualize intracellular microRNAs with low abundance.

Microplastics are solid polymer particles with a wide variety of surface properties, found in most waterbodies, and known as carriers of distinct microbial communities affecting the fate of the particles in the environment. Little is known about the formation of mineral deposits on microplastics and how these deposits connect to microbial assemblages and affect the physicochemical properties of the particles. In addition, most of the available research on this topic is based on large microplastics with sizes between 100 μm and up to 5 mm, rather than the small microplastics often found in drinking water sources. To narrow this gap in our understanding of environmental effects on small microplastics, two types of small microplastics made of two distinct polymers, poly(methyl methacrylate) (PMMA) and poly(tetrafluoroethylene) (PTFE) with sizes ranging from 15 to 150 μm, were incubated for six months in unprocessed and processed drinking water with increasing ionic concentration to allow for the formation of mineral deposits and microbial assemblages. Spatially resolved analysis with fluorescent in situ hybridization and confocal Raman microscopic imaging revealed deposits of calcium carbonates and scattered microbial assemblages on all microplastics, with structure, extend, and microbial association with the carbonates depending on the respective microplastic. Notably, PTFE floatation was overcome after three months in unprocessed drinking water but remained unchanged in processed drinking water, whereas PMMA appeared unaffected, indicating that the fate of microplastics in the environment may depend on polymer type and the encountered aquatic conditions forming mineral and microbial attachments to the particle surface.

A smartphone-based analysis system is favored for point-of-care testing applications. The present work proposes a novel micro-spectroscopy/imaging system comprising a portable spectrometer as an optical sensor and a compact homemade microscope to acquire the image and spectra of micron-scale regions. Protein concentration quantification based on the bicinchoninic acid method was demonstrated with the proposed micro-spectroscopy/imaging system to analyse the spectrometer signals. Morphologies of onion endothelial and human breast cancer cells, used as biological sample models, were characterized to demonstrate the microscopic imaging capacity of the device. The ability to simultaneously obtain morphological and spectral information using the proposed portable device was demonstrated by examining the 10 μm sub-pixels of a smartphone screen. These results highlight the potential for adopting a smartphone-based micro-spectroscopy/imaging system for point-of-care testing.

Focusing performance is a major concern for systems based on hydrodynamic focusing. In this study, the hydrodynamic focusing subsystem of a microscopic imaging system was analysed and modelled. The theoretical model was used to analyse the velocity and distribution range of sample particles in the focused sample flow in the micro-channel of the hydrodynamic focusing subsystem, when the velocities of the sample and sheath flows were varied. The results were used to optimise the coupling velocities of the sample and sheath flows for the microscopic imaging system, to keep working efficiency and image quality of the system simultaneously. An independent experiment was then conducted for verification, and the results agreed well with the theoretical investigation. The results of this study provide a general framework for adjusting the sample and sheath flow velocities to optimise the hydrodynamic focusing performance.

Background Dormant chemotherapy-resistant leukemia cells can survive for an extended period before relapse. Nevertheless, the mechanisms underlying the development of chemoresistance in vivo remain unclear. Methods Using intravital bone imaging, we characterized the behavior of murine acute myeloid leukemia (AML) cells (C1498) in the bone marrow before and after chemotherapy with cytarabine. Results Proliferative C1498 cells exhibited high motility in the bone marrow. Cytarabine treatment impaired the motility of residual C1498 cells. However, C1498 cells regained their migration potential after relapse. RNA sequencing revealed that cytarabine treatment promoted MRTF-SRF pathway activation. MRTF inhibition using CCG-203971 augmented the anti-tumor effects of chemotherapy in our AML mouse model, as well as suppressed the migration of chemoresistant C1498 cells. Conclusions These results provide novel insight into the role of cell migration arrest on the development of chemoresistance in AML, as well as provide a strong rationale for the modulation of cellular motility as a therapeutic target for refractory AML.

Cellobiohydrolases directly convert crystalline cellulose into cellobiose and are of biotechnological interest to achieve efficient biomass utilization. As a result, much research in the field has focused on identifying cellobiohydrolases that are very fast. Cellobiohydrolase A from the bacterium Cellulomonas fimi (CfCel6B) and cellobiohydrolase II from the fungus Trichoderma reesei (TrCel6A) have similar catalytic domains (CDs) and show similar hydrolytic activity. However, TrCel6A and CfCel6B have different cellulose-binding domains (CBDs) and linkers: TrCel6A has a glycosylated peptide linker, whereas CfCel6B's linker consists of three fibronectin type 3 domains. We previously found that TrCel6A's linker plays an important role in increasing the binding rate constant to crystalline cellulose. However, it was not clear whether CfCel6B's linker has similar function. Here we analyze kinetic parameters of CfCel6B using single-molecule fluorescence imaging to compare CfCel6B and TrCel6A. We find that CBD is important for initial binding of CfCel6B, but the contribution of the linker to the binding rate constant or to the dissociation rate constant is minor. The crystal structure of the CfCel6B CD showed longer loops at the entrance and exit of the substrate-binding tunnel compared with TrCel6A CD, which results in higher processivity. Furthermore, CfCel6B CD showed not only fast surface diffusion but also slow processive movement, which is not observed in TrCel6A CD. Combined with the results of a phylogenetic tree analysis, we propose that bacterial cellobiohydrolases are designed to degrade crystalline cellulose using high-affinity CBD and high-processivity CD.

We previously proposed a three-dimensional microscopic imaging system for objects hidden behind scattering media using in-line phase-shift digital holography, which simultaneously captures amplitude and phase information. However, as the thickness of the scattering medium increases, the influence of scattering is enhanced, and the reconstructed image of the object behind the scattering medium deteriorates. In this paper, we report the evaluation of this image using a near-infrared light source with a wavelength of 780 nm that is capable of deep tissue penetration. A favorable microscopic image of the object behind the rat-skin sample of 912 μm-thickness was successfully reconstructed.

Degradation of commercially available CdTe and CIGS PV modules installed in the desert conditions, are not well understood, because they have been in operation for relatively short period compared to crystalline silicon PV technology. In this paper, we investigate the degradation rate of both these thin-film technologies using indoor characterization methods. The calculated annual degradation rate of CdTe technology (1.3-2.2 percent/year) is lower than CIGS (5.3-8.8 /year). The main cause of this degradation is the reduction in fill factor caused by formation of permanent shunts. These shunts were characterized using Electroluminescence and microscopic imaging.

The mouse xenograft model is one of the most widely used animal model for biomedicine research. It is vital to distinguish the cells from different species, especially for the spatial distribution information. However, the available strategies of species-specific detection are either inapplicable in situ or of low specificity. Here, we reported a method based on DAPI staining, which offers an effective, convenient way that accurately identifies human and mouse nuclei at single-cell level in situ. This method was proven to be effective in cell co-culture and tumor xenograft tissue section. Microscopic imaging results shows obvious DAPI plaques-like structures in mouse nuclei, but absent in human nuclei. Moreover, we found these structures are co-localized with mouse major satellite DNA, which is located pericentromere in mouse, but absent in human. Our study provides a high-performance method that can be widely used for distinguish human and mouse cell in situ.

The purpose of this study is to develop hyperspectral imaging (HSI) for automatic detection of head and neck cancer cells on histologic slides. A compact hyperspectral microscopic system is developed in this study. Histologic slides from 15 patients with squamous cell carcinoma (SCC) of the larynx and hypopharynx are imaged with the system. The proposed nuclei segmentation method based on principle component analysis (PCA) can extract most nuclei in the hyperspectral image without extracting other sub-cellular components. Both spectra-based support vector machine (SVM) and patch-based convolutional neural network (CNN) are used for nuclei classification. CNNs were trained with both hyperspectral images and pseudo RGB images of extracted nuclei, in order to evaluate the usefulness of extra information provided by hyperspectral imaging. The average accuracy of spectra-based SVM classification is 68%. The average AUC and average accuracy of the HSI patch-based CNN classification is 0.94 and 82.4%, respectively. The hyperspectral microscopic imaging and classification methods provide an automatic tool to aid pathologists in detecting SCC on histologic slides.

Transmitted light imaging is an important tool in biophysics for applications that include sample analysis, recording samples whose viability is compromised by high levels of illumination (e.g., live cell tracking), and finding regions of interest in a sample. Koehler transillumination is a powerful illumination method used in commercial microscopes; yet commercial Koehler condensers are expensive, are difficult to integrate into tabletop systems, and make learning the concepts of Koehler illumination difficult because of their closed-box nature. Here, we show a protocol to build a simple 4f Koehler illumination system that offers advantages with respect to commercial condensers in terms of simplicity, cost, and compatibility with tabletop systems, such as open-source light sheet fluorescence microscopes. We include step-by-step instructions that can be followed by advanced undergraduate or graduate students without experience in optics on how to align and assemble the illuminator, along with a list of the necessary parts for assembly. We also include supplemental material that describes 4 supporting educational activities students can conduct with the apparatus and helps in the understanding of key concepts relevant to Koehler illumination and optics. The performance of the system is comparable to that of commercial condensers and significantly better, in terms of illumination homogeneity and depth of field (optical sections are possible), than that of LED flashlights, such as those found in low-cost diagnostic devices and tabletop systems.

Background and aimsDysregulated lipid metabolism has emerged as one of the major risk factors of atherosclerosis. Presently, there is a consensus that oxidized LDL (oxLDL) promotes development of atherosclerosis and downstream chronic inflammatory responses. Due to the dynamic metabolic disposition of lipoprotein, conventional approach to purify bioactive lipids for subsequent comprehensive analysis has proven to be inadequate for elucidation of the oxidized lipids species accountable for pathophysiology of atherosclerotic lesions. Herein, we aimed to utilize a novel mass microscopic imaging technology, coupled with mass spectrometry (MS) to characterize oxidized lipids in atherosclerotic lesions. MethodsWe attempted to use MALDI-TOF-MS and iMScope to identify selected oxidized lipid targets and visualize their respective localizations in study models of atherosclerosis. ResultsBased on the MS analysis, detection of 7-K under positive ionization through product ion peak at m/z 383 [M+H-H2O] indicated the distinctive presence of targeted lipid within Cu2+-oxLDL and Cu2+-oxLDL loaded macrophage-like J774A.1 cell, along with other cholesterol oxidation products. Moreover, the application of two-dimensional iMScope has successfully visualized the localization of lipids in aortic atherosclerotic plaques of the Watanabe heritable hyperlipidemic (WHHL) rabbit. Distinctive lipid distribution profiles were observed in atherosclerotic lesions of different sizes, especially the localizations of lysoPCs in atherosclerotic plaques. ConclusionsTaken together, we believe that both MALDI-TOF-MS and iMScope metabolomics technology may offer a novel proposition for future pathophysiological studies of lipid metabolism in atherosclerosis.

In recent years, the use of microinjections has increased in life science and biotechnology fields, specific examples include artificial insemination and gene manipulation. Microinjections are mainly performed based on visual information, thus, the operator needs high-level skill because of the narrowness of the visual field. Additionally, microinjections are performed as the operator views a microscopic image on a display, the position of the display requires the operator to maintain an awkward posture throughout the procedure. In this study, we developed a microscopic image display apparatus for microinjections based on a view-expansive microscope. The prototype of the view-expansive microscope has problems related to the variations in brightness and focal blur that accompany changes in the optical path length and amount of reflected light. Therefore, we propose the use of a variable-focus device to expand the visual field and thus circumvent the above-mentioned problems. We evaluated the observable area of the system using this variable-focus device. We confirmed that the observable area is 261.4 and 13.9 times larger than that of a normal microscope and conventional view-expansive microscopic system, respectively. Finally, observations of mouse embryos were carried out by using the developed system. We confirmed that the microscopic images can be displayed on a head-mounted display in real time with the desired point and field sizes.

Microscopic imaging modalities can be classified into two categories: those that form contrast from external agents such as dyes, and label-free methods that generate contrast from the object’s unmodified structure. While label-free methods such as brightfield, phase contrast, or quantitative phase imaging (QPI) are substantially easier to use, as well as non-toxic, their lack of specificity leads many researchers to turn to labels for insights into biological processes, despite limitations due to photobleaching and phototoxicity. The label-free image may contain the structures of interest, but it is often difficult or time-consuming to distinguish these structures from their surroundings. Here we summarize our recent progress in shattering this tradeoff, by using machine learning to perform automated segmentation on label-free, intrinsic contrast, quantitative phase images.

With UC2 (You-See-Too) we present an inexpensive 3D-printed microscopy toolbox. The system is based on concepts of modular development, rapid-prototyping and all-time accessibility using widely available off-the-shelf optic and electronic components. We aim to democratize microscopy, reduce the reproduction crisis and enhance trust into science by making it available to everyone via an open-access public repository. Due to its versatility the aim is to boost creativity and non-conventional approaches. In this paper, we demonstrate a development cycle from basic blocks to different microscopic techniques. First, we build a bright-field system and stress-test it by observing macrophage cell differentiation, apoptosis and proliferation incubator-enclosed for seven days with automatic focussing to minimize axial drift. We prove versatility by assembling a system using the same components to a fully working fluorescence light-sheet system and acquire a 3D volume of a GFP-expressing living drosophila larvae. Finally, we sketch and demonstrate further possible setups to draw a picture on how the system can be used for reproducible prototyping in scientific research. All design files for replicating the experimental setups are provided via an open-access online repository (https://github.com/bionanoimaging/UC2-GIT) to foster widespread use.

Despite an ever increasing demand for reliable and cheap methods in the detection and quantification of microbes, surprisingly few investigations have explored or utilized the luminescence of rare earths in the microbial context, neither in conventional, that is, plating and microscopic imaging techniques, nor in advanced methods like fluorescence flow cytometry. We have thus investigated the potential of some rare earth complexes and hybrid materials for microbiological analysis. We found fairly simple procedures for internal staining (dyes inside the bacterial cell) and external staining (dyes on the cell surface). The present paper is predominantly relying on microscopic imaging and luminescence spectroscopies (excitation, emission, decay times), but also evaluates model rare earth microspheres to estimate an eventual rare earth based stain for a fast and sensitive bacteria enumeration with luminescence flow cytometry.

The development of high throughput three-dimensional (3D) microscopic imaging technique is important for studying cell physiology and early-stage disease diagnoses. Here we propose and demonstrate a digital micromirror device (DMD) based angle-multiplexed high-speed optical diffraction tomography (ODT) technique. Using this ODT technique, we have achieved 3D imaging of cells at over 600 tomogram/second speed, which is 10-100 times faster than current ODTbased 3D cell imaging techniques. We envision that this high-speed ODT system will enable many cutting-edge biomedical applications, such as capturing millisecond scale cell dynamics in 3D space and high throughput 3D imaging of large cell populations.

PurposeThe aim of this study was to evaluate the retreatment abilities of the ProTaper Next (PTN) and F6 SkyTaper (F6) systems by using micro-computed tomography (micro-ct), radiographic and microscopic imaging techniques.Materials and MethodsThe root canals of twenty-six extracted mandibular premolar teeth were prepared and obturated. For the retreatment procedure, the teeth were randomly divided into two equal groups according to endodontic instruments: PTN (X4) and F6 (#40/.06). Pre- and post-operative filling material volumes were measured with micro-ct, and areas were measured with radiographic and microscopic imaging techniques. The percentages of residual material were calculated, and then statistically compared. The significance level was set at p0.05). PTN demonstrated better cleaning ability when evaluated by microscopic imaging. (p

The aim of the study was to analytically evaluate quantum dots in immunohistofluorescence (IHF-QD) microscopic imaging as detectors of food allergens&mdash, peanut and wheat. The experiment was designed as two in silico experiments or simulations: (a) models of pastry samples were prepared with the addition of allergenic components (peanut and wheat protein components) and without the addition of allergenic components, and (b) positive and negative commercial samples underwent food allergen detection. The samples from both simulations were tested by the ELISA and IHF-QD microscopic methods. The primary antibodies (secondary antibodies to a rabbit Fc fragment with labeled CdSe/ZnS QD) were labelled at 525, 585, and 655 nm emissions. The use of quantum dots (QDs) has expanded to many science areas and they are also finding use in food allergen detection, as shown in the study. The study indicated that differences between the ELISA and IHF-QD microscopic methods were not observable among experimentally produced pastry samples with and without allergenic components, although differences were observed among commercial samples. The important value of the study is certainly the differences found in the application of different QD conjugates (525, 585, and 655). The highest contrast was found in the application of 585 QD conjugates that can serve for the possible quantification of present food allergens&mdash, peanuts and wheat. The study clearly emphasized that QD can be used for the qualitative detection of food allergens and can represent a reliable analytical method for food allergen detection in different food matrixes.

Cell Separation is important in various biomedical fields. We have prepared gold nanoparticle (AuNP)-embedded collagen gels as a visible-light-responsive cell scaffold in which photoinduced single cell detachment occurs through local thermal denaturation of the collagen gel via the photothermal effect of AuNP. Physicochemical properties of collagen materials depend on the origin of the collagen and the presence of telopeptides. In this study, we prepared various AuNP-embedded collagen gels by using different collagen materials with and without the telopeptides to compare their thermal denaturation properties and photoinduced single cell detachment behaviors. Cellmatrix type I-C without telopeptides exhibited a lower denaturation temperature than Cellmatrix type I-A and Atelocell IAC, as examined by Fourier transform infrared (FTIR) spectroscopy, rheological analysis, and sol&ndash, gel transition observation. Three-dimensional (3D) laser microscopic imaging revealed that collagen fibers shrank in Cellmatrix type I-A upon heating, but collagen fibers disappeared in Cellmatrix type I-C upon heating. Cells cultured on the Cellmatrix type I-C-based AuNP-embedded collagen gel detached with shorter photoirradiation than on the Cellmatrix type I-A-based AuNP-embedded collagen gel, suggesting that collagen gels without telopeptides are suitable for a photoinduced single cell detachment system.

We describe a head fixation system allowing chronic microscopic imaging through cranial windows and optically transparent implanted microelectrode arrays in awake mice.

Since 1886, when the first picture of the human retina was taken, ocular imaging has played a crucial role in the diagnosis and management of ophthalmic diseases. One of the biggest contributors to the advancement of ocular imaging is the adoption of optical imaging techniques. Optical imaging is a method of looking into the body in a noninvasive way, like X-rays. However, unlike radiological imaging techniques that use ionizing radiation, optical imaging uses light and the properties of photons to produce detailed images ranging from structures as small as cells and molecules to structures as large as tissues and organs. There are plenty of advantages of using optical imaging compared to radiological imaging. For one, optical imaging is much safer for patients since it uses nonionizing radiation to excite electrons without causing damage. Additionally, since it is fast and safe, optical imaging can be used to monitor acute and chronic diseases, as well as treatment outcomes. Optical imaging is also useful for imaging soft tissue since different types of tissues absorb and scatter light differently. Finally, optical imaging can advantageously use varying colors of light to see and measure multiple properties of tissues at a time. Therefore, it is no surprise that the optical imaging modalities of fundus photography in the 1920s, scanning laser ophthalmoscope (SLO) imaging in 1981, and optical coherence tomography (OCT) in 1991 have touted a “golden age” in ophthalmic imaging applications. A new technology, photoacoustic imaging, has been shown to have promising features that could make it the next major imaging technique in ophthalmology. Additionally, photoacoustic imaging (PAI) can combine with preexisting optical microscopic imaging modalities to achieve multimodal imaging of the eye. In this chapter, we present a brief overview of fundus photography, SLO, and OCT while discussing the potential of PAI as the next major ocular imaging modality.

Background The aim of the study is to report the experience of the pathologists of the Italian Group for the Study of Inflammatory Bowel Disease (IBD) (group formed by pathologists with various experience) on the morphological assessment of digital slides pertaining to IBD and Non-IBD colitis underlining the necessity to implement this tool in daily routine and its utility to share opinions on difficult cases. Materials and Methods Forty-eight histological slides stained with haematoxylin and eosin obtained from ileo-colorectal endoscopic biopsies were digitized using Menarini D-Sight 2.0 system, uploaded onto a website platform and shared among 40 pathologists participating in the study. Information regarding the site of biopsy was disclosed; clinical data were blinded. Each participant was committed to write a comment on microscopic features purposing diagnostic opinion. One month after the last uploaded case, a form was sent to each participant to evaluate the personal experience on digital slide sharing. Results Sixteen pathologists out of 40 (40%) had consistently accessed to the site,9/40 (22%) commented on all slides, a diagnostic opinion was rendered in 8 slides. Most common critical issues were: A) poor internet connection resulting in ineffective evaluation of the digital slides, B) time-consuming cases raising difficult diagnostic interpretation, C) lack of clinical history. Overall, 24 participants (60%) found the forum valuable for practical training and educational purposes. Conclusions Sharing scanned slides circulating within a dedicated forum is an effective educational tool in both IBDs and Non-IBDs colitis. Although our results demonstrated a substantial compliance of the participants, their limited participation was an objective shortcoming. Hence, further efforts are needed to encourage this potentially rewarding practice among the pathologist community.

Disease diagnosis in low-resource settings is often hampered by lack of equipment and trained personnel required for microscopic examination of the biological sample. Recently, many smartphone-based microscopes have been developed and shown promise in microscopic imaging of the biological sample with minimal sample preparation. In this chapter, we will review the latest development of the smartphone microscopes. Various smartphone-based microscopy imaging methods will be reviewed. Approaches for smartphone microscope objective lenses will be discussed for their advantages and challenges. Finally, remaining challenges in translating the smartphone microscopy devices into field diagnostic tools will be discussed.

We report the use of a plasmonic random laser as an illumination source for microscopic imaging. The high correlation coefficient (0.91) quantifies the random laser’s ability to preserve information from disruptions caused by coherent artefacts.

In the pharmaceutical industry, high-throughput crystallization (HTC) is an emerging strategy to accelerate the discovery of active pharmaceutical ingredients (APIs) with appropriate crystallization properties. It is typically processed in 96- and 384-well plates, which offer parallel crystallization vessels with microliter-scaled volume. However, the microwell size effect on crystallization kinetics at the scale of microtiters has not been fully investigated and remains unsolved. To address the issue, we proposed a novel deep-learning-based approach to investigate the effects of microwell sizes on the crystal growth kinetics of APIs. With a combination of microscopic imaging and deep learning, the massive information of indomethacin crystals was successfully obtained and analyzed in a high-throughput manner. And the relationships between crystallization properties (the average size, crystal size distribution, and the growth rate of crystals) and the size of the crystallization vessels were successfully revealed, which were further validated by the crystallization of ibuprofen and the scale-up experiment. Our research demonstrated the powerful capability of deep learning in processing complicated crystalline images. Moreover, our findings in the relationships between the size of the crystallization vessels and crystal growth kinetics are of great value in guiding the manufacture of APIs in the pharmaceutical industry.

Deep learning has shown the high potential for the use of resolution enhancement in microscopic imaging. However, a large amount of paired datasets are usually required for training the networks. Data preparation is costly from the microscopic experiment. In this paper, the cycle generative adversarial network, trained by only a small amount of unpaired experimental microscopic images, is used to generate paired data for supervising learning in resolution enhancement. A typical network for resolution enhancement, named Pix2Pix-GAN, is then trained by the generated images to generate high-resolution images. A various of samples have been tested. From the images obtained with low-power microscopes, the trained network can output the images similar to those acquired with high-power microscopes. Results show that the generated dataset can effectively replace the experimental one for neural network training. Problems related to the experimental paired data preparation are then solved. The proposed method further broadens the application of deep learning for the optical field, especially for resolution enhancement in dynamic microscopic imaging.

The use of automated techniques for image analysis of microscopic wood specimens together with new procedures for the preparation of stained xylem tissue support the use of quantitative wood anatomy. These techniques and procedures are especially useful in the studies of retrospective analysis of xylem phenology, reaction(s) of trees to stressful conditions of growth, or reconstruction of long-term growth trends. The unresolved technical problems during the digitalization of cross sections from entire increment cores were stabilization and precise shifting of long microscopic specimens onto the optical microscope stage. For this reason, we have developed a long slide holder for microscope stages in two versions: the basic one allowing stabilization and manual shifting, and the advanced one for stabilization and mechanical shifting. Both versions of the adapter speed up the work with long slides, improving the quality of panoramic images of microscopic specimens.

Computational ghost imaging (CGI) presents a new way to solve the problem of extended depth of field (DOF), which has been studied for a long time. The application of correlation theory confirms that a plane light source can be used to avoid the limitation of DOF in CGI. Results from experiments where real objects are imaged by CGI between 0.3 m and 1.2 m verify that images are consistent in quality. This confirms that CGI is not affected by DOF and can be used to improve the accuracy of integral and microscopic imaging.

Single-molecule and -particle imaging with optical microscopic methods have been widely used in various fields with the advantages of high resolution, high throughput and excellent sensitivity. This review summarized recent progresses of optical microscopic imaging of single-molecule and -particle in the areas of molecular detection, super-resolution imaging and single-particle catalysis. At last, we outlined the challenges and prospect of optical microscopic imaging techniques for single-molecule and -particle assay in biomedical applications.

Analysis of red organic pigments in artworks (and in forensics applications) is challenging, because conventional nondestructive mapping techniques provide little contrast, and most chemical analyses with high specificity require sample removal. Here we demonstrate a new optical approach, pump-probe microscopy, for the analysis of red organic pigments. We investigate Carmine naccarat, Lac dye, purpurin, alizarin, madder lake, and eosin Y and show that their intrinsic photophysical properties produce distinctive pump-probe spectra. We utilize this contrast for high-resolution, three-dimensional imaging without the need for physical sample removal. Lastly, we highlight the potential of pump-probe microscopy as an analytical tool for forensics of other types of organic colorants by investigating a series of automotive paints.