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Investigations of factors influencing cell division and cell shape in bacteria are commonly performed in conjunction with high-resolution fluorescence microscopy as observations made at a population level may not truly reflect what occurs at a single cell level. Live-cell timelapse microscopy allows investigators to monitor the changes in cell division or cell morphology which provide valuable insights regarding subcellular localization of proteins and timing of gene expression, as it happens, to potentially aid in answering important biological questions. Here, we describe our protocol to monitor phenotypic changes in Bacillus subtilis and Staphylococcus aureus using a high-resolution deconvolution microscope. The objective of this report is to provide a simple and clear protocol that can be adopted by other investigators interested in conducting fluorescence microscopy experiments to study different biological processes in bacteria as well as other organisms.
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http://dx.doi.org/10.3791/59905 | DOI Listing |
Methods
September 2025
Gynaecology and Obstetrics, The Second Affiliated Hospital of Harbin Medical University, Harbin Medical University, Heilongjiang 150081, PR China. Electronic address:
Single-cell surface-enhanced Raman scattering (SERS) has emerged as a powerful tool for precision medicine owing to its label-free detection, ultrasensitivity, and unique molecular fingerprinting. Unlike conventional bulk analysis, it enables detailed characterization of cellular heterogeneity, with particular promise in circulating tumor cell (CTC) identification, tumor microenvironment (TME) metabolic profiling, subcellular imaging, and drug sensitivity assessment. Coupled with microfluidic droplet systems, SERS supports high-throughput single-cell analysis and multiparametric screening, while integration with complementary modalities such as fluorescence microscopy and mass spectrometry enhances temporal and spatial resolution for monitoring live cells.
View Article and Find Full Text PDFNeurobiol Aging
September 2025
O-Force Co., Ltd., 3454 Irino Kuroshio-cho, Hata-gun, Kochi 789-1931, Japan; Department of Pharmacology, Kochi Medical School, Kochi University, Kohasu, Oko-cho, Nankoku, Kochi 783-8505, Japan. Electronic address:
Due to the growing number of Alzheimer's disease (AD) patients, new drugs are urgently required. A synthetic nonapeptide, JAL-TA9 (YKGSGFRMI), derived from Transducer of ErbB-2.1 (Tob1) protein, cleaves amyloid β (Aβ) 42 with serine protease-like activity.
View Article and Find Full Text PDFJ Am Chem Soc
September 2025
Life-like Materials and Systems, University of Mainz, Duesbergweg 10-14, 55128 Mainz, Germany.
Transmembrane signaling is essential for cellular communication, yet reconstituting such mechanisms in synthetic systems remains challenging. Here, we report a simple and robust DNA-based mechanism for transmembrane signaling in synthetic cells using cholesterol-modified single-stranded DNA (Chol-ssDNA). We discovered that anchored Chol-ssDNA spontaneously flips across the membrane of giant unilamellar lipid vesicles (GUVs) in a nucleation-driven, defect-mediated process.
View Article and Find Full Text PDFAngew Chem Int Ed Engl
September 2025
Department of Physics and Chemistry, DGIST, Daegu, 42988, Republic of Korea.
Investigation of the fundamental microscopic processes occurring in organic reactions is essential for optimising both organocatalysts and synthetic strategies. In this study, single-molecule fluorescence microscopy was employed to study the Diels-Alder reaction catalysed by a first-generation MacMillan catalyst, providing direct insights into its kinetic dynamics. This reaction proceeds via a series of reversible processes under equilibrium conditions (S ⇄ IM ⇄ IM → P, IM and IM: N,O-acetal and iminium ion intermediates, respectively).
View Article and Find Full Text PDFLuminescence
September 2025
Department of Physics, Saveetha Engineering College (Autonomous), Chennai, Tamilnadu, India.
The iron nickel magnesium tetra-oxide (FeNiMgO) nanocomposites (NCs) first reported in this article were synthesized using the sol-gel method. For investigation using powder X-ray diffraction (PXRD), the presence of a cubic structure is confirmed. In Raman spectroscopy, the vibrational modes are investigated.
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