Measuring and imaging diffusion with multiple scan speed image correlation spectroscopy

• Verfasst von: Gröner, Nadine [VerfasserIn]
• Verfasst von: Capoulade, Jérémie [VerfasserIn]
• Verfasst von: Cremer, Christoph [VerfasserIn]
• Verfasst von: Wachsmuth, Malte [VerfasserIn]
• Titel: Measuring and imaging diffusion with multiple scan speed image correlation spectroscopy
• Verf.angabe: Nadine Gröner, Jérémie Capoulade, Christoph Cremer, and Malte Wachsmuth
• E-Jahr: 2010
• Jahr: 22 Sep 2010
• Umfang: 13 S.
• Fussnoten: Gesehen am 26.05.2023
• Titel Quelle: Enthalten in: Optics express
• Ort Quelle: Washington, DC : Optica, 1997
• Jahr Quelle: 2010
• Band/Heft Quelle: 18(2010), 20, Seite 21225-21237
• ISSN Quelle: 1094-4087
• DOI: doi:10.1364/OE.18.021225
• Datenträger: Online-Ressource
• Sprache: eng
• Sach-SW: Confocal laser scanning microscopy
• Sach-SW: Diffraction limit
• Sach-SW: Fluorescence microscopy
• Sach-SW: Fluorescence spectroscopy
• Sach-SW: Image processing
• Sach-SW: Temporal resolution
• K10plus-PPN: 1846255864

Abstract

The intracellular mobility of biomolecules is determined by transport and diffusion as well as molecular interactions and is crucial for many processes in living cells. Methods of fluorescence microscopy like confocal laser scanning microscopy (CLSM) can be used to characterize the intracellular distribution of fluorescently labeled biomolecules. Fluorescence correlation spectroscopy (FCS) is used to describe diffusion, transport and photo-physical processes quantitatively. As an alternative to FCS, spatially resolved measurements of mobilities can be implemented using a CLSM by utilizing the spatio-temporal information inscribed into the image by the scan process, referred to as raster image correlation spectroscopy (RICS). Here we present and discuss an extended approach, multiple scan speed image correlation spectroscopy (msICS), which benefits from the advantages of RICS, i.e. the use of widely available instrumentation and the extraction of spatially resolved mobility information, without the need of a priori knowledge of diffusion properties. In addition, msICS covers a broad dynamic range, generates correlation data comparable to FCS measurements, and allows to derive two-dimensional maps of diffusion coefficients. We show the applicability of msICS to fluorophores in solution and to free EGFP in living cells.