A versatile dual spot laser scanning confocal microscopy system for advanced fluorescence correlation spectroscopy analysis in living cell.
A fluorescence correlation spectroscopy 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.
Ferrand
Patrick
P
Institut Fresnel, Aix-Marseille Université, Campus de St Jérôme, F-13013 Marseille, France. patrick.ferrand@fresnel.fr
Pianta
Martina
M
Kress
Alla
A
Aillaud
Alexandre
A
Rigneault
Hervé
H
Marguet
Didier
D
D000465:Algorithms; D000818:Animals; D019556:COS Cells; D002138:Calibration; D002522:Chlorocebus aethiops; D049452:Green Fluorescent Proteins / Q000235:genetics; D018613:Microscopy, Confocal / Q000295:instrumentation / Q000379:methods; D012235:Rhodamines / Q000737:chemistry; D013050:Spectrometry, Fluorescence / Q000295:instrumentation / Q000379:methods; D018800:Thy-1 Antigens / Q000378:metabolism; D013997:Time Factors
D012235:Rhodamines; D018800:Thy-1 Antigens; C026188:rhodamine 6G; D049452:Green Fluorescent Proteins