@article{9ef41e93d1784dabbec9a9b6667b35f6,
title = "A Biophysics Toolbox for Reliable Data Acquisition and Processing in Integrated Force-Confocal Fluorescence Microscopy",
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.",
author = "Zhaowei Liu and \{van Veen\}, Edo and Humberto S{\'a}nchez and Bel{\'e}n Solano and \{Palmero Moya\}, \{Francisco J.\} and McCluskey, \{Kaley A.\} and \{Ram{\'i}rez Montero\}, Daniel and \{van Laar\}, Theo and Dekker, \{Nynke H.\}",
year = "2024",
month = apr,
day = "17",
doi = "10.1021/acsphotonics.3c01739",
language = "English",
volume = "11",
pages = "1592--1603",
journal = "ACS Photonics",
issn = "2330-4022",
publisher = "American Chemical Society",
number = "4",
}