- 标题
- 摘要
- 关键词
- 实验方案
- 产品
-
Subcellular chemical imaging of structurally similar acridine drugs by near-field laser desorption/laser postionization mass spectrometry
摘要： Insights into the pharmacologic effect on cellular processes and the potential toxicological effects are vital to new drug development and evaluation, yet research on these subjects remains a great challenge due to the lack of information regarding the spatiotemporal distribution of drugs and metabolites within a single cell. Mass spectrometry imaging (MSI) has proven to be a label-free and high-throughput approach for visualizing drug distribution in spatial and temporal domains. However, single-cell drug imaging has been limited so far by detection sensitivity and microscale lateral resolution. Herein, we report near-field laser desorption/laser postionization mass spectrometry (NDPI-MS) for single-cell imaging of two structurally similar drugs, proflavine and ethacridine, and subcellular distributions of proflavine at different drug concentrations were investigated. The NDPI-MS imaging results indicate that proflavine was accumulated in lysosomes, which was verified by laser scanning confocal microscopy (LSCM). Additionally, a distinguished subcellular distribution pattern of ethacridine from proflavine could be visualized, highlighting the complexity of the interaction between the drugs and biological environment even though these two drugs possess similar structures. Taken together, the present results demonstrate the great potential of the integrated single-cell MSI platform for characterizing the drug distribution and its phenotype changes within individual cells, expediting the identification and evaluation of newly developed drugs.
关键词： single cell imaging,mass spectrometry,acridine drugs,laser postionization,near-field
更新于2025-09-23 15:19:57
-
Detection of Metabolic Changes Induced via Drug Treatments in Live Cancer Cells and Tissue Using Raman Imaging Microscopy
摘要： Isocitrate dehydrogenase 1 (IDH1) mutations in gliomas, fibrosarcoma, and other cancers leads to a novel metabolite, D-2-hydroxyglutarate, which is proposed to cause tumorigenesis. The production of this metabolite also causes vulnerabilities in cellular metabolism, such as lowering NADPH levels. To exploit this vulnerability, we treated glioma and fibrosarcoma cells that harbor an IDH1 mutation with an inhibitor of nicotinamide adenine dinucleotide (NAD+) salvage pathway, FK866, and observed decreased viability in these cells. To understand the mechanism of action by which the inhibitor FK866 works, we used Raman imaging microscopy and identified that proteins and lipids are decreased upon treatment with the drug. Raman imaging showed a different distribution of lipids throughout the cell in the presence of the drug compared with the untreated cells. We employed nuclear magnetic resonance NMR spectroscopy and mass spectrometry to identify the classes of lipids altered. Our combined analyses point to a decrease in cell division due to loss of lipid content that contributes to membrane formation in the in vitro setting. However, the FK866 drug did not have the same potency in vivo. The use of Raman imaging microscopy indicated an opposite trend of lipid distribution in the tissue collected from treated versus untreated mice when compared with the cells. These results demonstrate the role of Raman imaging microscopy to identify and quantify metabolic changes in cancer cells and tissue.
关键词： NAD+ synthesis,tissue imaging,single cell imaging,microscopy,Raman spectrometry,fibrosarcoma IDH1
更新于2025-09-19 17:15:36
-
Correlated Secondary Ion Mass Spectrometry-Laser Scanning Confocal Microscopy Imaging for Single Cell-Principles and Applications
摘要： Secondary ion mass spectrometry (SIMS) as a powerful surface analysis technique has been widely applied in semiconductor industry and geology research. Recently, with the development of instrumental technology, SIMS is attracting more and more attention in life sciences. SIMS can provide surface MS spectra, 2D/3D chemical images and depth profiling of substances simultaneously. The minimal lateral resolution of 2D SIMS imaging is 80?100 nm, and the longitudinal resolution of 3D SIMS imaging is about 1–5 nm. However, owing to lack of specific ions to render the structures of organelles, SIMS imaging for single cells still have great challenges. Optical microscopy, in particular laser scanning confocal microscopy (LSCM), has been emerged to be an indispensable technique for single cell imaging and can obtain high spatial 2D/3D imaging to visualize the structures of organelles. Thus, the combinational use of SIMS and LSCM, which takes advantages of SIMS for molecular imaging and LSCM for morphological imaging, has greatly extended the application of SIMS imaging and ensured its accuracy at single cells level, providing novel insights into better understanding of the biological events inside cells. In this review, we focus on the development and application of SIMS imaging and the correlated SIMS and LSCM imaging in the research of cell biology and drug discovery. We anticipate that the combinational use of SIMS and LSCM imaging has promising future in biomedicine and life sciences.
关键词： Cell biology,Single cell imaging,Laser scanning confocal microscopy,Correlated secondary ion mass spectrometry and laser scanning confocal microscopy imaging,Secondary ion mass spectrometry,Review
更新于2025-09-16 10:30:52
-
Confined electrochemiluminescence in vertically ordered silica mesochannels for the imaging of hydrogen peroxide released from single cells
摘要： Here, the confined electrochemiluminescence (ECL) of luminol and hydrogen peroxide is achieved in vertically ordered silica mesochannels (SMCs) and applied to map the efflux of hydrogen peroxide from single living cells. The vertical alignment of SMCs on indium tin oxide (ITO) slides restricts the lateral diffusion of species generated in the ECL process. Accordingly, the cross-talk of luminescence intensity from nearby microregions is minimized for higher-spatial-resolution ECL imaging. The characterization of luminol ECL on these SMC-coated slides yields a detection range of hydrogen peroxide between 5 μM and 1 mM, providing high sensitivity in imaging the efflux of hydrogen peroxide from cells. Compared with the luminescence from individual cells on bare ITO slides, ~ 4-fold intensity is observed for SMC-coated ITO, suggesting a better electrode surface for single-cell ECL imaging.
关键词： single-cell imaging,hydrogen peroxide,silica mesochannels,confined electrochemiluminescence,luminol
更新于2025-09-10 09:29:36
-
Optical sectioning in multifoci Raman hyperspectral imaging
摘要： In this study, we compared the depth discrimination and speed performance of multifoci Raman hyperspectral imaging with the reference standard of a single laser point confocal Raman mapping. A liquid crystal spatial light modulator was employed for the generation of multifoci laser beams, and a digital micromirror device was used as a software‐configurable reflective pinhole array. The patterns of the laser foci and pinhole array can be rapidly changed without requiring any hardware alterations. Confocal patterns with different distance‐to‐size ratios were tested and compared. After optimization of the laser‐foci pattern, we demonstrated the feasibility of multifoci Raman hyperspectral microscopy for recording depth‐resolved molecular maps of biological cells (Acanthamoeba castellanii trophozoites). Micrometric depth discrimination and short acquisition times (20 min for single plane confocal image) were achieved.
关键词： single cell imaging,cross‐talk,confocal Raman imaging,multi‐beam,optical sectioning
更新于2025-09-04 15:30:14