Evaluation of optical properties of fluorescent nanofiber using image-processing technique

Purpose The purpose of this study is evaluate the optical properties of polyacrylonitrile (PAN) nanofibers containing fluorescent agents such as fluorescent dye and carbon quantum dots (CQDs) by using image-processing technique of Fluorescence microscope image. Design/methodology/approach The fluorescence microscope image of the pure PAN, PAN/CQDs and PAN/fluorescent dye nanofibers composite was analyzed using several image-processing techniques such as color histogram, lookup table (LUT), Fourier transform, RGB profile and surface plot analysis. Findings The fluorescence microscope image indicates that the fluorescence emission of nanocomposites depends on the type of fluorescent agent. The fluorescence intensity of nanofiber containing CQDs is more than nanofiber containing fluorescent dye. Various image-processing methods provide similar results for optical property of nanocomposites. Analyzing the LUT, the blue value of CQDs/PAN nanocomposite image was significantly higher than other nanocomposites. This was confirmed by other methods such as Fourier transform, color histogram and 3D topography of the electrospun nanofibers. According to analysis of colorimetric parameters, higher negative value of b* indicates bluer color for CQDs/PAN nanofibers than other nanocomposites. The obtained results indicate that the image-processing technique can be used to evaluate the optical property of fluorescent nanocomposite. Originality/value This study evaluates the optical properties of fluorescent nanocomposites by using image-processing techniques such as Fourier transform, color histogram, RGB profiles, LUT, surface plot and histogram analysis.

Morphology and Rheology of a Cool-Gel (Protein) Blended with a Thermo-Gel (Hydroxypropyl Methylcellulose)

This study investigates the morphological and rheological properties of blended gelatin (GA; a cooling-induced gel (cool-gel)) and hydroxypropyl methylcellulose (HPMC; a heating-induced gel (thermo-gel)) systems using a fluorescence microscope, small angle X-ray scattering (SAXS), and a rheometer. The results clearly indicate that the two biopolymers are immiscible and have low compatibility. Moreover, the rheological behavior and morphology of the GA/HPMC blends significantly depend on the blending ratio and concentration. Higher polysaccharide contents decrease the gelling temperature and improve the gel viscoelasticity character of GA/HPMC blended gels. The SAXS results reveal that the correlation length (ξ) of the blended gels decreases from 5.16 to 1.89 nm as the HPMC concentration increases from 1 to 6%, which suggests that much denser networks are formed in blended gels with higher HPMC concentrations. Overall, the data reported herein indicate that the gel properties of gelatin can be enhanced by blending with a heating-induced gel.

Effect of radiofrequency therapy on HPV16-E7 lentivirus infection in the reproductive tract of mice and its effect on immune function of splenic lymphocytes

Objective: To investigate the effect of radiofrequency therapy (RFT) on HPV16-E7 lentivirus infection in the reproductive tract of mice and reveal its effect on immune function of splenic lymphocytes. Materials and Methods: The mouse reproductive tract model was established by infection with HPV16-E7 lentivirus. Fluorescence microscope was used to evaluate successful injection. The expression of HPV16-E7 protein was detected by Western blotting test. The levels of CD4+ and CD8+ were determined by flow cytometry, and the ratio was calculated. The proliferation of splenic lymphocytes was detected by MTT assay. Expression of Interleukin (IL)-2 and interferon-γ (IFN-γ) messenger RNA (mRNA) in lymphocyte was determined by reverse transcription-quantitative polymerase chain reaction (RT-qPCR). Results: Fluorescence microscope determined the successful injection of HPV16-E7 lentivirus. Compared with model group, RFT treatment decreased HPV16-E7 protein, and increased CD4+/CD8+ ratio and the proliferation activity of splenic lymphocytes. Besides, RFT treatment increased the mRNA expression levels of IL-2 and IFN-γ compared to the model group. In particular, the proliferation activity of spleen lymphocytes and the expression levels of IL-2 mRNA and IFN-γ mRNA in RFT were higher at 12 days than at 6 days after treatment. Conclusion: RFT could eliminate HPV16-E7 lentivirus infection in the reproductive tract of mice, and the mechanism was related to the immune system.

Polyamidoamine-Remdesivir Conjugate: Physical Stability and Cellular Uptake Enhancement

Nucleoside analogue antiviral remdesivir works by inhibiting the RNA-dependent RNA polymerase enzyme and terminating the viral replication. Currently, remdesivir is under a clinical trial for its activity against SARS-CoV-2. In the blood, remdesivir will undergo an enzymatic reaction to become monophosphate analogue form which is difficult to penetrate into the cell membrane. PAMAM (polyamidoamine) dendrimer is a good carrier to encapsulate remdesivir as a water-insoluble drug (0,339 mg/mL). Entrapment of remdesivir in the PAMAM cavity avoided remdesivir molecules to not undergo the enzymatic reactions. This study aimed to synthesize, characterize and evaluate cellular uptake of PAMAM-Remdesivir conjugate. PAMAM-Remdesivir was prepared with various stirring times (3, 6, 12, 24, and 48 hours). The conjugates were characterized to observe the size and particle distribution using Particle Size Analyzer, encapsulating efficiency using UV-Vis Spectroscopy, interaction between PAMAM and remdesivir particle using Fourier Transform Infrared Spectroscopy and cellular uptake of PAMAM-RDV using Fluorescence Microscope. The optimized stirring time of PAMAM-Remdesivir conjugate was 24 hours wich resulted the particles charge of + 23,07 mV of zeta potential, 1008 nm of particle size, 0,730 of PDI, and 69% entrapment efficiency. In addition, the FTIR analysis showed that remdesivir molecules successfully conjugated to PAMAM. Thus, through strring optimization time, the remdesivir molecules were successfully entrapped to PAMAM cavity. The cellular uptake in Vero Cell of PAMAM-RDV conjugated fluorescein isothiocyanate was observed with fluorescence microscope and had a stronger intensity than remdesivir only solution.

Fluorescence microscope (Zeiss) v1

How to operate the fluorescence microscope in the lab. Beware of safety hazards: This instrument uses led radiation. Do not stare at the operating lamp for a prolonged period of time in fluorescence mode.

Preliminary Report: Rapid Intraoperative Detection of Residual Glioma Cell in Resection Cavity Walls Using a Compact Fluorescence Microscope

Objective: The surgical eradication of malignant glioma cells is theoretically impossible. Therefore, reducing the number of remaining tumor cells around the brain–tumor interface (BTI) is crucial for achieving satisfactory clinical results. The usefulness of fluorescence–guided resection for the treatment of malignant glioma was recently reported, but the detection of infiltrating tumor cells in the BTI using a surgical microscope is not realistic. Therefore, we have developed an intraoperative rapid fluorescence cytology system, and exploratorily evaluated its clinical feasibility for the management of malignant glioma. Materials and methods: A total of 25 selected patients with malignant glioma (newly diagnosed: 17; recurrent: 8) underwent surgical resection under photodiagnosis using photosensitizer Talaporfin sodium and a semiconductor laser. Intraoperatively, a crush smear preparation was made from a tiny amount of tumor tissue, and the fluorescence emitted upon 620/660 nm excitation was evaluated rapidly using a compact fluorescence microscope in the operating theater. Results: Fluorescence intensities of tumor tissues measured using a surgical microscope correlated with the tumor cell densities of tissues evaluated by measuring the red fluorescence emitted from the cytoplasm of tumor cells using a fluorescence microscope. A “weak fluorescence” indicated a reduction in the tumor cell density, whereas “no fluorescence” did not indicate the complete eradication of the tumor cells, but indicated that few tumor cells were emitting fluorescence. Conclusion: The rapid intraoperative detection of fluorescence from glioma cells using a compact fluorescence microscope was probably useful to evaluate the presence of tumor cells in the resection cavity walls, and could provide surgical implications for the more complete resection of malignant gliomas.