Print Email Facebook Twitter A Biophysics Toolbox for Reliable Data Acquisition and Processing in Integrated Force-Confocal Fluorescence Microscopy Title A Biophysics Toolbox for Reliable Data Acquisition and Processing in Integrated Force-Confocal Fluorescence Microscopy Author Liu, Z. (TU Delft BN/Nynke Dekker Lab; Kavli institute of nanoscience Delft) van Veen, E.N.W. (TU Delft BN/Nynke Dekker Lab; Kavli institute of nanoscience Delft) Sanchez González, H. (TU Delft BN/Nynke Dekker Lab; Kavli institute of nanoscience Delft) Solano Hermosilla, B.P. (TU Delft BN/Nynke Dekker Lab; Kavli institute of nanoscience Delft) Palmero Moya, F.J.P.M. (TU Delft BN/Nynke Dekker Lab; Kavli institute of nanoscience Delft) Mc Cluskey, K.A. (TU Delft BN/Nynke Dekker Lab; Kavli institute of nanoscience Delft) Ramirez Montero, D.F. (TU Delft BN/Nynke Dekker Lab; Kavli institute of nanoscience Delft) van Laar, T. (TU Delft BN/Nynke Dekker Lab; Kavli institute of nanoscience Delft) Dekker, N.H. (TU Delft BN/Nynke Dekker Lab; University of Oxford; Kavli Institute of Nanoscience Discovery) Date 2024 Abstract Integrated single-molecule force-fluorescence spectroscopy setups allow for simultaneous fluorescence imaging and mechanical force manipulation and measurements on individual molecules, providing comprehensive dynamic and spatiotemporal information. Dual-beam optical tweezers (OT) combined with a confocal scanning microscope form a force-fluorescence spectroscopy apparatus broadly used to investigate various biological processes, in particular, protein:DNA interactions. Such experiments typically involve imaging of fluorescently labeled proteins bound to DNA and force spectroscopy measurements of trapped individual DNA molecules. Here, we present a versatile state-of-the-art toolbox including the preparation of protein:DNA complex samples, design of a microfluidic flow cell incorporated with OT, automation of OT-confocal scanning measurements, and the development and implementation of a streamlined data analysis package for force and fluorescence spectroscopy data processing. Its components can be adapted to any commercialized or home-built dual-beam OT setup equipped with a confocal scanning microscope, which will facilitate single-molecule force-fluorescence spectroscopy studies on a large variety of biological systems. Subject automated data acquisitiondata analysisfluorescence spectroscopyforce spectroscopyoptical tweezersprotein−DNA interactions To reference this document use: http://resolver.tudelft.nl/uuid:338b1be6-8fff-4c7b-b524-2489c88199bf DOI https://doi.org/10.1021/acsphotonics.3c01739 ISSN 2330-4022 Source ACS Photonics, 11 (4), 1592-1603 Part of collection Institutional Repository Document type journal article Rights © 2024 Z. Liu, E.N.W. van Veen, H. Sanchez González, B.P. Solano Hermosilla, F.J.P.M. Palmero Moya, K.A. Mc Cluskey, D.F. Ramirez Montero, T. van Laar, N.H. Dekker Files PDF liu-et-al-2024-a-biophysi ... nfocal.pdf 3.87 MB Close viewer /islandora/object/uuid:338b1be6-8fff-4c7b-b524-2489c88199bf/datastream/OBJ/view