We describe proof-of-concept for a novel approach for visualizing regions of close apposition between the surfaces of living cells. A membrane-anchored protein with high affinity for a chemical ligand is expressed on the surface of one set of cells, and the cells are co-cultured with a second set of cells that express a membrane-anchored fluorogen-activating protein (FAP). The co-cultured cells are incubated with a bivalent reagent composed of fluorogen linked to the high-affinity ligand, with the concentration of the bivalent reagent chosen to be less than the binding constant for the FAP-fluorogen pair but greater than the binding constant for the ligand-high-affinity protein pair. In these conditions, strong FAP signal is observed only in regions of close proximity between membranes of the two classes of cell, where high local concentration of fluorogen favors binding to the FAP.
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http://dx.doi.org/10.1021/acs.bioconjchem.7b00047 | DOI Listing |
Genome Med
January 2025
Blizard Institute, Barts and The London Faculty of Medicine and Dentistry, Queen Mary University of London, London, E1 2AT, UK.
Background: Senescence classification is an acknowledged challenge within the field, as markers are cell-type and context dependent. Currently, multiple morphological and immunofluorescence markers are required. However, emerging scRNA-seq datasets have enabled an increased understanding of senescent cell heterogeneity.
View Article and Find Full Text PDFOpen Biol
January 2025
Department of Cell and Developmental Biology, University of Colorado Anschutz Medical Campus School of Medicine, Aurora, CO 80045, USA.
Protein-based nanomachines drive every cellular process. An explosion of high-resolution structures of multiprotein complexes has improved our understanding of what these machines look like and how they work, but we still know relatively little about how they assemble in living cells. For example, it has only recently been appreciated that many complexes assemble co-translationally, with at least one subunit still undergoing active translation while already interacting with other subunits.
View Article and Find Full Text PDFMethods Appl Fluoresc
January 2025
Guangxi Medical University, No. 22, Shuangyong Road, Qingxiu District, Nanning, Guangxi, Nanning, Guangxi, 530021, CHINA.
Cell viability assessment plays a crucial role in biological research, pharmaceutical development, and toxicological identification. Here, we used GelRed, a sensitive and safer nucleic acid dye, to selectively label dead cells with red fluorescence (FL) thus distinguishing dead cells from live ones. Further more, the combined use of GelRed and SYTO 9 (another nucleic acid dye) enabled the clear differentiation in FL spectra between the two physiological statuses.
View Article and Find Full Text PDFBiomed Phys Eng Express
January 2025
Xi'an Jiaotong University, No.28 Xianning West Road, Xi'an, Shaanxi 710049, P.R. China, Xi'an, 710049, CHINA.
The optimal method for three-dimensional thermal imaging within cells involves collecting intracellular temperature responses while simultaneously obtaining corresponding 3D positional information. Current temperature measurement techniques based on the photothermal properties of quantum dots face several limitations, including high cytotoxicity and low fluorescence quantum yields. These issues affect the normal metabolic processes of tumor cells.
View Article and Find Full Text PDFEnviron Sci Pollut Res Int
January 2025
Waste Science and Technology, Luleå University of Technology, Luleå, Sweden.
Improper management of wood impregnation chemicals and treated wood has led to soil contamination at many wood treatment sites, particularly with toxic substances like creosote oil and chromated copper arsenate (CCA). The simultaneous presence of these pollutants complicates the choice of soil remediation technologies, especially if they are to be applied in situ. In this laboratory study, we attempted to immobilise arsenic (As) and simultaneously degrade polycyclic aromatic hydrocarbons (PAHs) (constituents of creosote oil) by applying a modified electrochemical oxidation method.
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