Taxonomic significance of morphological and elemental characteristics of achenes of Artemisia genus from Turkey.

In the study, the achene macromorphological and micromorphological characters of the genus Artemisia distributed in Turkey have been researched with the target of knowing systematically important carpological structures for the examined species. Macro-morphological structures of the achenes including color, shape, dimension, and carpopodium diameter were studied with 100 achenes of 10 specimens per taxa with a Light Microscope. Micro-morphological features of the achenes containing surface ornamentation, anticlinal and periclinal cell walls, epidermal cells, and the presence of secondary structures were examined with a Scanning Electron Microscope. EDS analyses were performed with a SEM. EDS analyses were carried out by selecting the same spot on the sample surface at 80 sec under 30 µm aperture size, with 20 kV acceleration voltage, 8 mm operating distance, high current, and processing time conditions. The color, shape, and dimension of achene have macro-morphologically shown variations. The examined achenes are separated into four shapes; fusiform-oblong, oblong, oblong-ovate, and ovate. Oblong-ovate is the most common type. Achene dimensions range from 0.62 to 2.48 mm in length, and from 0.30 to 1.21 mm in width. Also, carpopodium diameter varies between 0.10 and 0.19 mm. Achene surfaces of the examined taxa are micro-morphologically assessed, and substantial differences are noticeably detected on behalf of the surface structures for instance, surface ornamentation, anticlinal and periclinal cell walls, epidermal cells, and the presence of secondary structures of the achenes. Surface ornamentation is separated into 10 types: irregularly sulcate, regularly sulcate, ruminate, sulcate-scalariform, rugose, favulariate, slightly sulcate, alveolate, tuberculate, and reticulate. A percentage comparison of the elements in the achene pericarp of the studied taxa has been performed with SEM-EDS. Accordingly, pericarps in taxa include C, Ca, K, Mg, Cl, Si, Na, and S elements. In the taxonomy of the genus Artemisia, the achene morphological characters are very significant characteristics that disclose inter-specific relations among the examined taxa. Moreover, a dichotomous key is offered for the identification of the studied taxa based on achene characters. RESEARCH HIGHLIGHTS: The achenes of Turkish Artemisia taxa have been examined in depth. The morphological characteristics of achenes of Turkish Artemisia taxa have been studied using SEM and LM for the first time and debated the systematic practice of these characters. The elemental content of the achene pericarp has been systematically evaluated for the first time.

Fluorescent microscopy evaluation of diode laser effect on the penetration depth of turmeric (Curcuma longa) extract cream on skin tissues of Wistar rats.

The study investigates the effect of diode laser exposure on curcumin's skin penetration, using turmeric extraction as a light-sensitive chemical and various laser light sources. It uses an in vivo skin analysis method on Wistar strain mice. The lasers are utilized at wavelengths of 403 nm, 523 nm, 661 nm, and 979 nm. The energy densities of the lasers are 20.566 J/cm2, 20.572 J/cm2, 21.162 J/cm2, and 21.298 J/cm2, which are comparable to one another. The experimental animals were divided into three groups: base cream (BC), turmeric extract cream (TEC), and the combination laser (L), BC, and TEC treatment group. Combination light source (LS) with cream (C) was performed with 8 combinations namely 523 nm ((L1 + BC) and (L1 + TEC)), 661 nm ((L2 + BC) and (L2 + TEC)), 403 nm ((L3 + BC) and (L3 + TEC)), and 979 nm ((L4 + BC) and (L4 + TEC)). The study involved applying four laser types to cream-covered and turmeric extract-coated rat skin, with samples scored for analysis. The study found that both base cream and curcumin cream had consistent pH values of 7-8, within the skin's range, and curcumin extract cream had lower viscosity. The results of the statistical analysis of Kruskal-Wallis showed a significant value (p < 0.05), which means that there are at least two different laser treatments. The results of the post hoc analysis with Mann-Whitney showed that there was no significant difference in the LS treatment with the addition of BC or TEC when compared to the BC or TEC treatment alone (p > 0.05), while the treatment using BC and TEC showed a significant difference (p < 0.05). Laser treatment affects the penetration of the turmeric extract cream into the rat skin tissue.

Application of microscopy and spectroscopy in investigating anti-cancer potential of Achyranthes aspera L. leaves.

The genus Achyranthes belong to the family Amaranthaceae which constitutes an important group of herbs and shrubs with immense medicinal value. The present research work was conducted to investigate the anticancer potential of Achyranthes aspera L. leaves by focusing on the antioxidant, aniproliferative and antimitotic activities of leaf extracts. Plant extraction was carried out by soxhelt method with different solvents. Phytochemical characterization of the plants extracts using chemical methods identified the presence of cardiac glycosides, saponins, coumarins, proteins, tannins, flavonoids and triterpenes. Alkaloid was present in methanolic and ethanolic extract. High performance liquid chromatography showed presence of different concentration of myricetin, quercetin and kaempferol in different extracts with the highest concentration of myricetin (84.53 µg/mL) in n-butanolic extract. The extracts were then tested for antioxidant activity using 2,2-diphenylpicrylhydrazyl (DPPH) radical scavenging assay by spectrophotometric method. In DPPH radical scavenging assay, antioxidant activity of A. aspera ranged between 79.78 ± 0.034% and 58.63 ± 0.069%. Highest antioxidant activity was observed for methanolic extract and lowest for acetone. Antimitotic activity was determined by using Allium cepa assay in which microscopic investigation was carried out to observe normal and abnormal phases of mitosis. In this assay, n-butanolic extract had highest antimitotic activity with minimum mitotic index at 2 mg/mL (57 ± 0.0351%). The plant extracts also caused chromosomal and mitotic aberrations which were clearly observed under 40× and 100× magnification of compound microscope. Antiproliferative activity was determined by using yeast cell model in which light microscope with hemocytometer was used for cell counting. In case of Antiproliferative activity, the ethyl acetate extract of A. aspera had highest antiproliferative activity with lowest cell viability (22.14 ± 0.076%) at highest extract concentration (2 mg/mL) while methanol extract of A. aspera had highest antiproliferative activity with lower cell viability (24.24 ± 0.057%) at lowest extract concentration (0.25 mg/mL). The results of the study indicated that the leaves extract of A. aspera have strong potential to be used as a source of anti-cancer agent. RESEARCH HIGHLIGHTS: Achyranthes aspera L. leaves have various phytochemicals which contribute to its medicinal properties Various extracts of the leaves of A. aspera L. possess antioxidant, antimitotic and antiproliferative potential The results of the study indicated that the leaves extract of A. aspera have strong potential to be used as a source of anti-cancer agent.

Boosting microscopic object detection via feature activation map guided poisson blending.

Microscopic examination of visible components based on micrographs is the gold standard for testing in biomedical research and clinical diagnosis. The application of object detection technology in bioimages not only improves the efficiency of the analyst but also provides decision support to ensure the objectivity and consistency of diagnosis. However, the lack of large annotated datasets is a significant impediment in rapidly deploying object detection models for microscopic formed elements detection. Standard augmentation methods used in object detection are not appropriate because they are prone to destroy the original micro-morphological information to produce counterintuitive micrographs, which is not conducive to build the trust of analysts in the intelligent system. Here, we propose a feature activation map-guided boosting mechanism dedicated to microscopic object detection to improve data efficiency. Our results show that the boosting mechanism provides solid gains in the object detection model deployed for microscopic formed elements detection. After image augmentation, the mean Average Precision (mAP) of baseline and strong baseline of the Chinese herbal medicine micrograph dataset are increased by 16.3% and 5.8% respectively. Similarly, on the urine sediment dataset, the boosting mechanism resulted in an improvement of 8.0% and 2.6% in mAP of the baseline and strong baseline maps respectively. Moreover, the method shows strong generalizability and can be easily integrated into any main-stream object detection model. The performance enhancement is interpretable, making it more suitable for microscopic biomedical applications.

Pollen variability in Quercus L. species and relative systematic implications.

This study aims to address one of the challenges related to the complexity of the Quercus L. genus, that is the identification of structural elements favouring the systematic identification of the oak pollen. Thus, in this contribution, we explored the variation of morphometric and chemical parameters in pollen samples collected from 47 different Quercus species and hybrids. Several qualitative (e.g., outline in polar view, class, aperture structures) and quantitative (e.g., diameter, exine and sporoderm thickness, autofluorescence, content in proteins and plant metabolites) features were evaluated by optic microscopy and spectrophotometric assays. Statistical analyses were also carried out to assess significant correlations and clustering effects among the studied taxa, based on phenotypical and biochemical data, to identify the parameters which could be useful for taxonomic discrimination at inter- and intra-specific level. Only few morphological traits showed the potentiality to be diagnostic, such as pollen diameter and outline in polar view. The intensity of pollen autofluorescence varied among the samples but it did not seem to correlate with protein, carotenoid, phenolic and flavonoid content. However, differences in protein and carotenoid levels were detected, suggesting them as possible taxonomic discriminants for oak pollen. Thus, our work represents a step forward in understanding morphology and biochemistry of oak pollen, constitutes an experimental set-up applicable in future systematic studies on other genera, and opens new perspectives for further molecular investigations on Quercus species.

Morphoanatomy, Histochemistry, and Phytochemistry of Stem and Leaves of Crateva tapia L.

Crateva tapia L. occurs only in Brazil and belongs to the family Capparaceae. This study aimed to characterize the morphoanatomy and histochemistry of stem and leaves, and phytochemical tests of the leaf of C. tapia. Macroscopic characterization was performed with the aid of a pachymeter and a stereomicroscope. Cross-sections of the stem, petiole, petiolule, and leaf blade, as well as paradermal sections of the leaf blade were mounted on microscope slides. Analyzes were carried out with optical microscopy. For histochemical analysis, different reagents were useda ccording to the targeted metabolite. Phytochemical tests of the methanolic extracts of the leaves were performed using thin layer chromatography. Crateva tapia has a stem with a circular cross-section and smooth bark. The leaves are alternate spirally, compound 3-foliolate. The leaf blade is elliptical and has a reticulate dictiodromous nerve. Through anatomical characterization, it was possible to identify a hypostomatic leaf blade with anisocytic, tetracytic, and anomocytic stomata and the presence of secretory canals. Through histochemistry and phytochemical tests, it was possible to observe the presence of different metabolites. Thus, the results obtained in this study contribute to the quality control of C. tapia, a species only found in Brazil, increasing the taxonomic knowledge of the family Capparaceae.

Authentication and Quality Control of the Brazilian Traditional Herb "Espinheira-Santa" (Monteverdia ilicifolia) by Morpho-Anatomy and Microscopy.

The leaves of Monteverdia ilicifolia (syn. Maytenus ilicifolia), commonly called espinheira-santa, are widely used in South American traditional medicines to treat gastritis and ulcers. Several products labeled as espinheira-santa are sold as dietary supplements in retail stores and via e-commerce. Many different species with similar leaf morphology are often mistaken for Monteverdia ilicifolia and used as espinheira-santa, including Monteverdia aquifolia (Celastraceae), Citronella gongonha (Cardiopteridaceae), Jodina rhombifolia (Santalaceae), Sorocea bonplandii (Moraceae), and Zollernia ilicifolia (Fabaceae). This study aimed to characterize M. ilicifolia and distinguish it from adulterants using morphological and microscopic techniques. In addition, foreign matter and powder characteristics of botanical materials sold as "espinheira-santa" were analyzed. The morphoanatomical studies of the leaves and stems of M. ilicifolia and its five adulterant species have revealed noteworthy features that can help species identification and quality control of commercial espinheira-santa. This study showed that many commercial espinheira-santa materials were adulterated and of inferior quality.

An Image-Based High-Throughput and High-Content Drug Screening Method Based on Microarray and Expansion Microscopy.

A high-efficiency drug screening method is urgently needed due to the expanding number of potential targets and the extremely long time required to assess them. To date, high throughput and high content have not been successfully combined in image-based drug screening, which is the main obstacle to improve the efficiency. Here, we establish a high-throughput and high-content drug screening method by preparing a superhydrophobic microwell array plate (SMAP) and combining it with protein-retention expansion microscopy (proExM). Primarily, we described a flexible method to prepare the SMAP based on photolithography. Cells were cultured in the SMAP and treated with different drugs using a microcolumn-microwell sandwiching technology. After drug treatment, proExM was applied to realize super-resolution imaging. As a demonstration, a 7 × 7 image array of microtubules was successfully collected within 3 h with 68 nm resolution using this method. Qualitative and quantitative analyses of microtubule and mitochondria morphological changes after drug treatment suggested that more details were revealed after applying proExM, demonstrating the successful combination of high throughput and high content.

High-field magnetic resonance microscopy of aortic plaques in a mouse model of atherosclerosis.

OBJECTIVES: Pre-clinical models of human atherosclerosis are extensively used; however, traditional histological methods do not allow for a holistic view of vascular lesions. We describe an ex-vivo, high-resolution MRI method that allows the 3 dimensional imaging of the vessel for aortic plaque visualization and quantification. MATERIALS AND METHODS: Aortas from apolipoprotein-E-deficient (apoE-/-) mice fed an atherogenic diet (group 1) or a control diet (group 2) were subjected to 14 T MR imaging using a 3D gradient echo sequence. The obtained data sets were reconstructed (Matlab), segmented, and analyzed (Avizo). The aortas were further sectioned and subjected to traditional histological analysis (Oil-Red O and hematoxylin staining) for comparison. RESULTS: A resolution up to 15 × 10x10 µm3 revealed that plaque burden (mm3) was significantly (p < 0.05) higher in group 1 (0.41 ± 0.25, n = 4) than in group 2 (0.01 ± 0.01, n = 3). The achieved resolution provided similar detail on the plaque and the vessel wall morphology compared with histology. Digital image segmentation of the aorta's lumen, plaque, and wall offered three-dimensional visualizations of the entire, intact aortas. DISCUSSION: 14 T MR microscopy provided histology-like details of pathologically relevant vascular lesions. This work may provide the path research needs to take to enable plaque characterization in clinical applications.

Label-free drug interaction screening via Raman microscopy.

Development of a simple, label-free screening technique capable of precisely and directly sensing interaction-in-solution over a size range from small molecules to large proteins such as antibodies could offer an important tool for researchers and pharmaceutical companies in the field of drug development. In this work, we present a thermostable Raman interaction profiling (TRIP) technique that facilitates low-concentration and low-dose screening of binding between protein and ligand in physiologically relevant conditions. TRIP was applied to eight protein-ligand systems, and produced reproducible high-resolution Raman measurements, which were analyzed by principal component analysis. TRIP was able to resolve time-depending binding between 2,4-dinitrophenol and transthyretin, and analyze biologically relevant SARS-CoV-2 spike-antibody interactions. Mixtures of the spike receptor-binding domain with neutralizing, nonbinding, or binding but nonneutralizing antibodies revealed distinct and reproducible Raman signals. TRIP holds promise for the future developments of high-throughput drug screening and real-time binding measurements between protein and drug.

Study on identification algorithm of traditional Chinese medicinals microscopic image based on convolutional neural network.

When the similarity of medicinal materials is high and easily confused, the traditional subjective judgment has an impact on the identification results. Use high-dimensional features to identify medicinal materials to ensure the quality of Chinese herbal concoction products and proprietary Chinese medicines. OBJECTIVE: To study the identification algorithm of traditional Chinese medicinals (TCM) microscopic images based on convolutional neural network (CNN) to improve the objectivity and accuracy of microscopic image identification of TCM powders. METHODS: Microscopic image datasets of 4 TCM powders sclereids of Rhizoma Coptidis, Cortex Magnoliae Officinalis, Cortex Phellodendri Chinensis, and Cortex Cinnamomi were constructed, and 400 collected images, as the model training and testing objects, were identified and classified by AlexNet model, VGGNet-16, VGGNet-19, and GoogLeNet model. RESULTS: The average recognition accuracy in the tested microscopic image of AlexNet model, VGGNet-16, VGGNet-19, and the GoogLeNet model is 93.50%, 95.75%, 95.75%, and 97.50% correspondingly. CONCLUSION: The GoogLeNet model has a higher classification accuracy and is the best model to achieve real-time. Applying the CNN to the identification of microscopic images of TCM powders makes the operation of TCM identification simpler and the measurement more accurate while improving repeatability.

Calcium Imaging in Mouse Superior Colliculus.

The superior colliculus (SC), an evolutionarily conserved midbrain structure in all vertebrates, is the most sophisticated visual center before the emergence of the cerebral cortex. It receives direct inputs from ~30 types of retinal ganglion cells (RGCs), with each encoding a specific visual feature. It remains elusive whether the SC simply inherits retinal features or if additional and potentially de novo processing occurs in the SC. To reveal the neural coding of visual information in the SC, we provide here a detailed protocol to optically record visual responses with two complementary methods in awake mice. One method uses two-photon microscopy to image calcium activity at single-cell resolution without ablating the overlaying cortex, while the other uses wide-field microscopy to image the whole SC of a mutant mouse whose cortex is largely undeveloped. This protocol details these two methods, including animal preparation, viral injection, headplate implantation, plug implantation, data acquisition, and data analysis. The representative results show that the two-photon calcium imaging reveals visually evoked neuronal responses at single-cell resolution, and the wide-field calcium imaging reveals neural activity across the entire SC. By combining these two methods, one can reveal the neural coding in the SC at different scales, and such combination can also be applied to other brain regions.

Clinical Validation of Stimulated Raman Histology for Rapid Intraoperative Diagnosis of Central Nervous System Tumors.

Stimulated Raman histology (SRH) is an ex vivo optical imaging method that enables microscopic examination of fresh tissue intraoperatively. The conventional intraoperative method uses frozen section analysis, which is labor and time intensive, introduces artifacts that limit diagnostic accuracy, and consumes tissue. SRH imaging allows rapid microscopic imaging of fresh tissue, avoids tissue loss, and enables remote telepathology review. This improves access to expert neuropathology consultation in both low- and high-resource practices. We clinically validated SRH by performing a blinded, retrospective two-arm telepathology study to clinically validate SRH for telepathology at our institution. Using surgical specimens from 47 subjects, we generated a data set composed of 47 SRH images and 47 matched whole slide images (WSIs) of formalin-fixed, paraffin-embedded tissue stained with hematoxylin and eosin, with associated intraoperative clinicoradiologic information and structured diagnostic questions. We compared diagnostic concordance between WSI and SRH-rendered diagnoses. Also, we compared the 1-year median turnaround time (TAT) of intraoperative conventional neuropathology frozen sections with prospectively rendered SRH-telepathology TAT. All SRH images were of sufficient quality for diagnostic review. A review of SRH images showed high accuracy in distinguishing glial from nonglial tumors (96.5% SRH vs 98% WSIs) and predicting final diagnosis (85.9% SRH vs 93.1% WSIs). SRH-based diagnosis and WSI-permanent section diagnosis had high concordance (κ = 0.76). The median TAT for prospectively SRH-rendered diagnosis was 3.7 minutes, approximately 10-fold shorter than the median frozen section TAT (31 minutes). The SRH-imaging procedure did not affect ancillary studies. SRH generates diagnostic virtual histologic images with accuracy comparable to conventional hematoxylin and eosin-based methods in a rapid manner. Our study represents the largest and most rigorous clinical validation of SRH to date. It supports the feasibility of implementing SRH as a rapid method for intraoperative diagnosis complementary to conventional pathology laboratory methods.

Authentication of Single Herbal Powders Enabled by Microscopy-Guided In Situ Auto-sampling Combined with Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry.

In the long history of investigation of herbal products, microscopic examination has greatly contributed to the authentication of herbs in a powder form. However, it cannot provide the chemical profiles of herbal powders and thus is limited to morphological identification. In this work, we present a label-free and automatic approach for the characterization and identification of single herbal powders and their adulterants, enabled through the combination of microscopy-guided auto-sampling and matrix-assisted laser desorption/ionization mass spectrometry (MALDI MS). To meet the demand for automatic and highly efficient in situ extraction, the glass slide was coated with gelatin to immobilize dried herbal powders that cannot stick to the glass slide like fresh and hydrated cells. The gelatin coating also facilitated the pump-out of chemical components and prevented diffusion across the interface enabled by the formation of a tight contact at the probe tip and surface. Optical microscopy was applied to acquire the microstructure and position of the herbal powders immobilized on the gelatin-coated slide. The candidate single herbal powders were picked out by a software for subsequent auto-sampling and MALDI MS identification. The combination of microstructure features and chemical profiles significantly improved the authentication capability of microscopic examination.

Oleofoams stabilized by monoacylglycerides: Impact of chain length and concentration.

Oleofoams are plant oil based whipped systems which have drawn academic and industry attention in recent years. The aim of this study was to determine the effect of fatty acid chain length and monoacylglyceride (MAG) concentration on the performance and structural properties of MAG-based oleofoams. Four different MAGs (monolaurin, monomyrystin, monopalmitin, and monostearin) were studied at three concentration levels (5, 10, and 15 wt%). The fatty acid chain length had a statistically significant impact on the size and shape of crystals formed, while higher MAG concentrations led to higher numbers of crystals in the continuous oil phase. These differences affected the performance and physical properties of the oleofoams: compared to other MAGs, monostearin based oleofoams were harder and exhibited higher values of G' and G″, had higher overrun and showed better stability. Lastly, through microscopy techniques it was successfully proved that monostearin-based oleofoams are stabilized by both bulk and Pickering stabilization.

The effect of herbicides on morphological features of pollen grains in Prunus serotina Ehrh. in the context of elimination of this invasive species from European forests.

Prunus serotina Ehrh. is an alien invasive neophyte widespread in European forests. So far, no effective methods of its elimination have been developed. For this reason, the aim of our study was to determine how herbicides affect the morphological characteristics of pollen grains. This knowledge may be crucial to control this invasive species. The current study was carried out in a research area of 2.7 ha located in the Zielonka Forest near Poznan, Poland (N 52°31'58.016″, E 17°05'55.588″). We tested morphological differences among ten features of P. serotina pollen, based on the samples collected from 15 control trees compared to the 50 trees treated with five different herbicides. In total 1950 pollen grains were measured. We confirmed the adopted hypotheses of long-term herbicide influence on P. serotina pollen. Pollen grains from the control trees had a longer equatorial axis, were more elongated in shape and had the largest range of exine thickness compared to the pollen from the herbicide-treated samples. Exine thickness in the control sample was on average 0.74 µm, ranging from 0.42 to 1.19 µm. The average values and the ranges of this trait in the samples treated with herbicides were larger (e.g. average exine thickness was from 0.90 to 0.95 µm). There were differences in the P/E ranges of variability between the control and herbicide-treated samples. In the control sample the P/E ratio was 1.32-2.04 and elongated forms of pollen shapes prevailed, while in the herbicide-treated samples it ranged from 1.03 to 1.47. The share of deformed pollen grains in the herbicide-treated samples was lower than expected, ranging from 8.7 to 25.3%, while in the control samples it was 6%. Logo and Mustang turned out to be the most effective among the herbicides used in the described research. The two used application methods were found to have an effect on pollen quality.

The applicability of Fourier transform infrared microspectroscopy for correction against matrix effects in X-ray fluorescence microimaging of tissues.

X-ray fluorescence (XRF) and Fourier transform infrared (FTIR) microscopy techniques are now considered popular for rapid and label-free complementary spectrochemical analysis of chemical elements and molecular systems in biological specimens. The morphological heterogeneity but also the inhomogeneities associated with the thickness/density of biological samples demonstrate challenges for the quantitative XRF microimaging. Therefore, in the present work, we proposed for the first time the application of the total absorbance under the FTIR spectra as a mass surface correction procedure for two-dimensional (2D) XRF microimaging of tissues. We also evaluated the equivalence of the developed correction method based on total absorbance of FTIR spectra with the proposed approaches based on incoherent scattering of primary X-rays as well as on the membrane Si-Kα transmission signal, on the example of selected rat organ tissues. Thin cryo-sections taken from various organs of Wistar rats were deposited on silicon nitride membranes (Si3N4). The FTIR microscopy studies were performed to collect infrared absorption spectra, used then for the determination of total absorbance values in the selected areas of tissue samples. In turn, hard X-ray imaging based on synchrotron radiation allowed the determination of characteristic radiation intensities of the elements detectable from the tissue, as well as the characteristic radiation of the membrane Si and incoherently scattered X-ray photons (Compton scattering). The latter served as correction factors for the surface mass of the sample alongside the FTIR total absorbance. The qualitative and quantitative analyses showed a high agreement between the results of elemental surface mass correction using total absorbance under FTIR spectra of tissues with those obtained using surface mass correction factors determined directly from XRF spectra. Therefore, the proposed procedure is a good alternative in cases where the surface mass effect of the sample cannot be eliminated based on the information provided directly by the XRF spectrum, as in the case of using polymer films as sample support. We have also proposed a procedure for synchronizing SRXRF and FTIR images, not limited to visual inspection of imaging/mapping data, but also enabling quantitative analysis. We found that the total absorbance determined from FTIR spectra can be successfully used as a correction factor for eliminating the surface mass effect in XRF microimaging of thin freeze-fried tissues and therefore to obtain the surface mass-independent elemental quantities. The proposed approach for 2D-FTIR-XRF analysis can also be a powerful and versatile tool for fostering a correlation and co-localization analysis to search for common distribution patterns between molecular arrangements and chemical elements.

Study on the application of optical coherence microscopy in Hirschsprung's disease.

To explore the clinical application value of optical coherence microscopy (OCM) in Hirschsprung's disease. 109 HSCR patients were recuited in a Chinese hospital from January 2018 to July 2021. All the recruited patients underwent barium enema angiography preoperatively and the resected diseased intestinal tubes were evaluated intraoperatively. The OCM and the histopathological examination were performed successively on the surgical specimens, and the OCM images were compared with the relevant tissue sections to characterize different lesions. 10 non-HSCR fetal colorectal tissues at the same period were retained for OCM, the characteristics of which with and without HSCR under OCM imaging were analyzed. In the OCM images of in vitro tissue, it can be clearly observed that the scattering degree of HSCR narrow segment mucosal is high, glands and crypt structures are reduced or even atrophy, and the scattering degree of submucosal and intermuscular is low; In the dilated segment, the low scattering and high scattering are complex, and the muscle layer is obviously hypertrophy and structural disorder. Compared with the pathological findings, the OCM sensitivity, Kappa value, and AUC area reached 92.66%, 0.63, and 0.91, respectively. OCM can quickly and clearly display the structure of all layers of colorectal tissue, which is highly consistent with the corresponding histopathological examination results and has high sensitivity. which will provide a more reliable basis for OCM diagnosis of early HSCR, targeted biopsy and location of operative treatment, and has a certain potential for clinical application.

A biophysical study of tear film lipid layer model membranes.

The tear film lipid layer (TFLL), the final layer of the human tear film is responsible for surface tension reduction while blinking, water evaporation retardation and maintaining the stability of the tear film. The study of the composition-structure-function relationship of TFLL is paramount, as a compromised structure of TFLL leads to the emergence of dry eye disease (DED) which is one the most prevalent ophthalmic surface diseases of the modern world, associated with chronic pain and reduced visual capability. In this model membrane study, a systematic approach is used to study the biophysical properties of TFLL model membranes as a function of composition. Three mixed-lipid model membranes are studied along with their individual components comprising cholesteryl oleate (CO), glyceryl trioleate (GT), L-α-phosphatidylcholine (egg PC) and a free fatty acid mixture. The models become progressively more complex from binary to quaternary mixtures, allowing the role of each individual lipid to be derived. Langmuir balance, Brewster Angle Microscopy (BAM) and Profile Analysis Tensiometer (PAT) are used to study the surface activity and compression-expansion cycles, morphology, and rheological behaviour of the model membranes, respectively. Evidence of multilayering is observed with inclusion of CO and a reversible collapse is associated with the GT phase transition. An initially more coherent film is observed due to the addition of polar PC. Notably, these individual behaviours are retained in the mixed films and suggest a possible role for each physiological component of TFLL.

Multi-microscopy techniques combined with FT-IR spectroscopy reveals the histological and biochemical causes leading to fruit texture difference in oriental melon (Cucumis melo var. Makuwa Makino).

Multi-microscopy techniques and Fourier transform infrared (FT-IR) spectroscopy were used in this study to investigate the intrinsic causes leading to fruit texture difference between two cultivars of oriental melon 'HDB' (crisp) and 'HPM' (mealy). On the histological aspect, orderly arranged regular-shaped cells with tissue natural fracture pattern showed cell rupture in 'HDB' versus loosely arranged irregular-shaped cells with tissue natural fracture pattern showed cell-to-cell separation in 'HPM' of sarcocarp are histological causes for crisp and mealy fruit texture, respectively. On the biochemical aspect, FT-IR spectra (4000-850 cm-1) of sarcocarp tissue cell wall materials (CWM) happened a dramatic change at the mature stage in 'HPM', but not in 'HDB'. Insightly, the lower de-methyl-esterified homogalacturonan (HG) abundance with higher water-soluble pectin (WSP) ratio and lower hemicellulose (HC) content contribute a poor intercellular adhesion in 'HPM' middle lamella (ML) at the mature stage compared to 'HDB'.