Structured Illumination Microscopy of Mitochondrial in Mouse Hepatocytes with an Improved Image Reconstruction Algorithm.

In this paper, a structured illumination microscopy (SIM) image reconstruction algorithm combined with notch function (N-SIM) is proposed. This method suppresses the defocus signal in the imaging process by processing the low-frequency signal of the image. The existing super-resolution image reconstruction algorithm produces streak artifacts caused by defocus signal. The experimental results show that the algorithm proposed in our study can well suppress the streak artifacts caused by defocused signals during the imaging process without losing the effective information of the image. The image reconstruction algorithm is used to analyze the mouse hepatocytes, and the image processing tool developed by MATLAB is applied to identify, detect and count the reconstructed images of mitochondria and lipid droplets, respectively. It is found that the mitochondrial activity in oxidative stress induced growth inhibitor 1 (OSGIN1) overexpressed mouse hepatocytes is higher than that in normal cells, and the interaction with lipid droplets is more obvious. This paper provides a reliable subcellular observation platform, which is very meaningful for biomedical work.

Aptamer/antibody sandwich method for digital detection of SARS-CoV2 nucleocapsid protein.

Nucleocapsid protein (N protein) is the most abundant protein in SARS-CoV2 and is highly conserved, and there are no homologous proteins in the human body, making it an ideal biomarker for the early diagnosis of SARS-CoV2. However, early detection of clinical specimens for SARS-CoV2 remains a challenge due to false-negative results with viral RNA and host antibodies based testing. In this manuscript, a microfluidic chip with femtoliter-sized wells was fabricated for the sensitive digital detection of N protein. Briefly, ß-galactosidase (ß-Gal)-linked antibody/N protein/aptamer immunocomplexes were formed on magnetic beads (MBs). Afterwards, the MBs and ß-Gal substrate fluorescein-di-ß-d-galactopyranoside (FDG) were injected into the chip together. Each well of the chip would only hold one MB as confined by the diameter of the wells. The MBs in the wells were sealed by fluorocarbon oil, which confines the fluorescent (FL) product generated from the reaction between ß-Gal and FDG in the individual femtoliter-sized well and creates a locally high concentration of the FL product. The FL images of the wells were acquired using a conventional inverted FL microscope. The number of FL wells with MBs (FL wells number) and the number of wells with MBs (MBs wells number) were counted, respectively. The percentage of FL wells was calculated by dividing (FL wells number) by (MBs wells number). The higher the percentage of FL wells, the higher the N protein concentration. The detection limit of this digital method for N protein was 33.28 pg/mL, which was 300 times lower than traditional double-antibody sandwich based enzyme-linked immunosorbent assay (ELISA).

Pigment analysis based on a line-scanning fluorescence hyperspectral imaging microscope combined with multivariate curve resolution.

A rapid and cost-effective system is vital for the detection of harmful algae that causes environmental problems in terms of water quality. The approach for algae detection was to capture images based on hyperspectral fluorescence imaging microscope by detecting specific fluorescence signatures. With the high degree of overlapping spectra of algae, the distribution of pigment in the region of interest was unknown according to a previous report. We propose an optimization method of multivariate curve resolution (MCR) to improve the performance of pigment analysis. The reconstruction image described location and concentration of the microalgae pigments. This result indicated the cyanobacterial pigment distribution and mapped the relative pigment content. In conclusion, with the advantage of acquiring two-dimensional images across a range of spectra, HSI conjoining spectral features with spatial information efficiently estimated specific features of harmful microalgae in MCR models.

Mechanical properties and molecular structure analysis of subsurface dentin after Er:YAG laser irradiation.

The purpose of this study was to evaluate the chemical and mechanical modifications in subsurface dentin layer after Er: YAG (Erbium-Yttrium Aluminium Garnet) laser irradiation, as the guidance of new dental restorative materials specific for laser irradiated dentin. MATERIALS AND METHODS: Dentin disks obtained from extracted human molars were prepared and exposed to a single pulse Er:YAG laser irradiation at 80mJ/pulse. After laser irradiation the mechanical and chemical characteristics of intertubular dentin in subsurface layer were studied using nanoindentation tester and micro-Raman spectromy (µ-RS). The dentin 5-50µm depth beneath the lased surface was determined as testing area. Two-way analysis of variance (ANOVA) were used to compare the mechanical values between lased and untreated subsurface dentin (P = 0.05). RESULTS: A laser affected subsurface dentin layer after Er:YAG laser treatment is present. The laser irradiation is considered to decrease the mechanical properties in the superficial subsurface layer (<15µm deep). There was no significant difference in nanohardness and Young's modulus between lased subsurface dentin and untreated dentin (p > 0.05) under the depth of 15µm. However, the dentin at 5µm and 10µm depth beneath the lased surface exhibited significantly lower (~ 47.8% and ~ 33.6% respectively) hardness (p < 0.05). Er:YAG laser irradiation affected both mineral and organic components in subsurface dentin layer, a higher degree of crystallinity and reduced organic compounds occurred in the lased subsurface dentin. CONCLUSION: Under the tested laser parameters, Er:YAG laser irradiation causes lower mechanical values and reduction of organic components in subsurface dentin, which has deleterious effects on resin adhesion to this area.