A versatile dual spot laser scanning confocal microscopy system for advanced fluorescence correlation spectroscopy analysis in living cell
P. Ferrand (IF), M. Pianta (IF), A. Kress (IF), A. Aillaud (IF), H., Rigneault (IF), D. Marguet (CIML)
TL;DR
This paper introduces a dual-spot confocal microscopy system enabling advanced fluorescence correlation spectroscopy in living cells, with independent control of measurement volumes and detailed protocols for optimized data acquisition.
Contribution
It presents a novel dual-spot FCS system with independent measurement volume control and a method to monitor excitation and collection efficiencies separately.
Findings
Demonstrated static and scanning measurements using the dual-spot system.
Provided detailed protocols for controlling measurement volume parameters.
Showcased the system's capability for advanced FCS analysis in living cells.
Abstract
A fluorescence correlation spectroscopy (FCS) system based on two independent measurement volumes is presented. The optical setup and data acquisition hardware are detailed, as well as a complete protocol to control the location, size and shape of the measurement volumes. A method that allows to monitor independently the excitation and collection efficiency distribution is proposed. Finally, a few examples of measurements that exploit the two spots in static and/or scanning schemes, are reported.
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