We present direct-LIVE-PAINT, an easy-to-implement approach for the nanoscopic imaging of protein structures in live cells using labeled binding peptides. We demonstrate the feasibility of direct-LIVE-PAINT with an actin-binding peptide fused to EGFP, the location of which can be accurately determined as it transiently binds to actin filaments. We show that direct-LIVE-PAINT can be used to image actin structures below the diffraction-limit of light and have used it to observe the dynamic nature of actin in live cells. We envisage a similar approach could be applied to imaging other proteins within live mammalian cells.
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http://dx.doi.org/10.1002/pro.4558 | DOI Listing |
Front Immunol
December 2024
Laboratory of Cellular and Molecular Biology, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD, United States.
Introduction: T cell activation requires T cell receptor (TCR) engagement by its specific ligand. This interaction initiates a series of proximal events including tyrosine phosphorylation of the CD3 and TCRζ chains, recruitment, and activation of the protein tyrosine kinases Lck and ZAP70, followed by recruitment of adapter and signaling proteins. CD28 co-stimulation is also required to generate a functional immune response.
View Article and Find Full Text PDFProbiotics Antimicrob Proteins
January 2025
Department of Cell and Molecular Biology and Microbiology, Faculty of Biological Science and Technology, University of Isfahan, Isfahan, Iran.
Heat-killed lactobacilli seem to have protective effects against oxidative stress and neurotoxicity. This study aimed to evaluate the antioxidant properties of specific heat-killed lactobacilli extracts and determine their neuroprotective effects against the neurotoxicity induced by blood plasma from people with multiple sclerosis (MS). The antioxidant activity of the three heat-killed lactobacilli was measured using the DPPH assay.
View Article and Find Full Text PDFJ Phys Chem Lett
January 2025
College of Physics and Optoelectronic Engineering, Key Laboratory of Optoelectronic Devices and Systems of Ministry of Education and Guangdong Province, State Key Laboratory of Radio Frequency Heterogeneous Integration (Shenzhen University), Shenzhen University, Shenzhen 518060, P. R. China.
One of the most significant advances in stimulated emission depletion (STED) super-resolution microscopy is its capacity for dynamic super-resolution imaging of living cells, including the long-term tracking of interactions between various cells or organelles. Consequently, the multicolor STED plays a pivotal role in biological research. Despite the emergence of numerous fluorescent probes characterized by low toxicity, high stability, high brightness, and exceptional specificity, enabling dynamic imaging of living cells with multicolor STED, practical implementation of multicolor STED for live-cell imaging is influenced by several factors.
View Article and Find Full Text PDFJ Phys Chem B
January 2025
OncoImmunin, Inc., 207A Perry Parkway, Suite 6, Gaithersburg, Maryland 20877, United States.
We have previously found that the presence of an H-type excitonic dimer formed by two fluorophores covalently bound to an oligonucleotide allows the delivery of such a polymer into live cells without inducing toxicity. We are now using time-resolved fluorescence measurements in solution to understand the molecular dynamics of an antisense probe and how pairing with complementary sense strands of various lengths and degrees of complementarity affects the antisense strand's properties. We report that a DNA strand composed of 30 residues and labeled with an H-type excitonic Cyanine-5/Cyanine-5 dimer shows a predominant 1.
View Article and Find Full Text PDFACS Appl Mater Interfaces
January 2025
State Key Laboratory of Solidification Processing, School of Materials Science and Engineering, Northwestern Polytechnical University (NPU), Xi'an 710072, China.
Gold nanoclusters (AuNCs) have garnered significant attention in biomedical applications, particularly in biosensing, cancer therapy, and imaging, due to their unique optical property, good biocompatibility, and distinct bioactivity. Understanding the cellular uptake behavior of AuNCs is critical to improve the efficacy of their applications, whose mechanism has not been adequately validated. In this work, we synthesized AuNCs with varying surface modifications to quantify the exact law of surface charge on the cellular uptake of AuNCs in a multidimensional manner by using 3D multicellular tumor spheroids of both HeLa cells and MCF-7 cells as the model system.
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