- 标题
- 摘要
- 关键词
- 实验方案
- 产品
-
Ultrasensitive U-shaped fiber optic LSPR cytosensing for label-free and in situ evaluation of cell surface N-glycan expression
摘要： Label-free and in situ detection of N-glycan expression on surface of cells is vital for understanding the progression of cancer. Herein, cytosensing based on U-shaped fiber optic local surface plasmon resonance (LSPR) was developed for cancer cell detection and N-glycan evaluation on cell surface. The U-shaped fiber optic LSPR has high refractive index sensitivity (RIS). Therefore, U-shaped fiber optic LSPR cytosensing afforded ultrasensitivity for cancer cell detection with the limit of detection (LOD) of 30 cells/mL and good linearity in a wide range of 1×102-1×106 cells/mL under the optimal conditions of modified AuNPs size and Con A concentration. The U-shaped fiber optic LSPR cytosensing shows more than 29 times lower limit of detection than the straight ones. More importantly, the prepared U-shaped cytosensing was applied to evaluate the N-glycan expression level on the surface of cancer cells under different concentration external stimuli of inhibitor tunicamycin (TM). Noticeably, U-shaped fiber optic LSPR cytosensing showed an acceptable reproducibility, satisfactory anti-interference and good selectivity, and was also applied in the assessment of N-glycan expression on surface of six kinds of cell lines. Therefore, the U-shaped fiber optic LSPR offers a feasible tool in biophysical research and clinical diagnosis for cancer.
关键词： Cancer cell,N-Glycan expression,Cytosensing,U-shaped fiber optic probe,Local surface plasmon resonance
更新于2025-09-23 15:23:52
-
Ultrasensitive and Label-Free Detection of Cell Surface Glycan Using Nanochannel-Ionchannel Hybrid Coupled with Electrochemical Detector
摘要： In this work, asymmetric nanochannel-ionchannel of porous anodic alumina (PAA) coupled with electrochemical detector was used for sensitive and label-free detection of cell surface glycan. The amplified ionic current caused by array nanochannels as well as the ionic current rectification (ICR) caused by asymmetric geometry endows PAA with sensitive ionic current response. Functionalized with the special molecular probe, the constructed nanofluidic device can be used for selective recognition and detection of glycan in real-time and label-free format. In addition, due to the subnanosize of ionchannels, the probe immobilization and glycan recognition is carried out on the outer surface of PAA, avoiding the blockage of PAA nanochannel by samples, which promises the reproducibility and accuracy of the present method toward bioanalysis. Results show that the glycan concentration ranging from 10 fM to 10 nM can be successfully detected with a detection limit of ~10 aM, which is substantially lower than most previous works. The designed strategy provides a valuable platform for sensitive and label-free detection of cell surface glycan, which acts as a promising candidate in pathological research and cancer diagnosis.
关键词： porous anodic alumina,ionic current rectification,cell surface glycan,nanochannel-ionchannel hybrid,nanofluidics
更新于2025-09-23 15:19:57
-
Quantum dots are conventionally applicable for wide-profiling of wall polymer distribution and destruction in diverse cells of rice
摘要： Plant cell walls represent enormous biomass resources for biofuels, and it thus becomes important to establish a sensitive and wide-applicable approach to visualize wall polymer distribution and destruction during plant growth and biomass process. Despite quantum dots (QDs) have been applied to label biological specimens, little is reported about its application in plant cell walls. Here, semiconductor QDs (CdSe/ZnS) were employed to label the secondary antibody directed to the epitopes of pectin or xylan, and sorted out the optimal conditions for visualizing two polysaccharides distribution in cell walls of rice stem. Meanwhile, the established QDs approach could simultaneously highlight wall polysaccharides and lignin co-localization in different cell types. Notably, this work demonstrated that the QDs labeling was sensitive to profile distinctive wall polymer destruction between alkali and acid pretreatments with stem tissues of rice. Hence, this study has provided a powerful tool to characterize wall polymer functions in plant growth and development in vivo, as well as their distinct roles during biomass process in vitro.
关键词： Glycan immunolabeling,Plant cell wall,Biomass,Rice,Quantum dots,Chemical pretreatment
更新于2025-09-19 17:13:59
-
Enhanced imaging of glycan expressing cancer cells using poly(glycidyl methacrylate)-grafted silica nanospheres labeled with quantum dots
摘要： Glycosylation on the cell surface contains abundant biological information, and detecting the glycan on cell surfaces can offer critical insight into biology and diseases. Here, a signal amplification strategy for the sensitive detection of glycan expression on the cell surface was proposed. In this approach, glycans on the cell surface were detected with poly(glycidyl methacrylate)-grafted silica nanosphere labeled with quantum dots (QDs) and biotin through the specific affinity reaction of avidin-biotin on the cancer cells. Glycans on the cell surface were first labeled via selective oxidization of sialyl groups into aldehydes by periodate. Aniline-catalyzed hydrazone ligation with biotin hydrazide was then used for the specific recognition to avidin. The nanoprobe was fabricated with "living" SiO2 nanoparticles with alkyl bromide groups on their surfaces. They were then subsequently grafted with poly(glycidyl methacrylate) (PGMA) brushes via the successive surface-initiated atom transfer radical polymerization. The CdTe QDs and biotin were immobilized through a ring-open reaction with epoxy groups in the PGMA brushes to obtain QDs/biotin-polymer brush-functionalized silica nanosphere (SiO2-PGMA-QDs/biotin). Enhanced sensitivity could be achieved by an increase in CdTe QDs loading per assay event, because of the large number of surface functional epoxy groups offered by the PGMA. As a result, fluorescence signal increased versus the unamplified method. This method successfully demonstrates a simple, specific, and potent method to detect glycans on the cell surface.
关键词： Glycan detection,Quantum dots,Polymer-brush,Imaging
更新于2025-09-12 10:27:22
-
Chemical derivatization coupled with matrix assisted laser desorption ionization time-of-flight mass spectrometry for convenient and efficient <i>N</i> -glycan detection in human plasma
摘要： As one of the most common post-translational modifications, protein N-glycosylation is involved in most important biological processes. Structural changes of the protein linked N-glycans have close correlations with the occurrence and progression of various diseases. Therefore, efficient detection of these disease-associated aberrations is of great significance for the development of new diagnostic biomarkers and therapeutic drug targets. In this work, we developed a method for convenient and effective N-glycan detection by coupling condition controlled chemical derivatization with MALDI-TOF MS. Due to the high reaction efficiency and enhanced hydrophobicity, improved identification scales were obtained for N-glycans from both asialofetuin and human plasma compared with the direct analysis.
关键词： asialofetuin,MALDI-TOF MS,N-glycan,human plasma,chemical derivatization
更新于2025-09-12 10:27:22
-
Egg white as a quality control in matrix assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI).
摘要： The strength of MALDI-MSI is to analyse and visualize spatial intensities of molecular features from an intact tissue. The distribution of the intensities can then be visualized within a single tissue section or compared in between sections, acquired consecutively. This method can be reliably used to reveal physiological structures and has the potential to identify molecular details, which correlate with biological outcomes. MALDI-MSI implementation in clinical laboratories requires the ability to ensure method quality and validation to meet diagnostic expectations. To be able to get consistent qualitative and quantitative results, standardized sample preparation and data acquisition are of highest priority. We have previously shown that the deposition of internal standards onto the tissue section during sample preparation can be used to improve mass accuracy of monitored m/z features across the sample. Here, we present the use of external and internal controls for the quality check of sample preparation and data acquisition, which is particularly relevant when either many spectra are acquired during a single MALDI-MSI experiment or data from independent experiments are processed together. To monitor detector performance and sample preparation, we use egg white as an external control for peptide and N-glycan MALDI-MSI throughout the experiment. We have also identified endogenous peptides from cytoskeletal proteins, which can be reliably monitored in gynecological tissue samples. Lastly, we summarize our standard quality control workflow designed to produce reliable and comparable MALDI-MSI data from single sections and tissue microarrays (TMAs).
关键词： egg white,MALDI-MSI,quality control,N-glycan,peptide,tissue microarrays,FFPE tissue
更新于2025-09-11 14:15:04