title: Development of new photon-counting detectors for single-molecule fluorescence microscopy.
abstract: Two optical configurations are commonly used in single-molecule fluorescence microscopy: point-like excitation and detection to study freely diffusing molecules, and wide field illumination and detection to study surface immobilized or slowly diffusing molecules. Both approaches have common features, but also differ in significant aspects. In particular, they use different detectors, which share some requirements but also have major technical differences. Currently, two types of detectors best fulfil the needs of each approach: single-photon-counting avalanche diodes (SPADs) for point-like detection, and electron-multiplying charge-coupled devices (EMCCDs) for wide field detection. However, there is room for improvements in both cases. The first configuration suffers from low throughput owing to the analysis of data from a single location. The second, on the other hand, is limited to relatively low frame rates and loses the benefit of single-photon-counting approaches. During the past few years, new developments in point-like and wide field detectors have started addressing some of these issues. Here, we describe our recent progresses towards increasing the throughput of single-molecule fluorescence spectroscopy in solution using parallel arrays of SPADs. We also discuss our development of large area photon-counting cameras achieving subnanosecond resolution for fluorescence lifetime imaging applications at the single-molecule level.
authors.affiliation: Department of Chemistry and Biochemistry, UCLA, Los Angeles, CA 90095-1547, USA. michalet@chem.ucla.edu
authors.forename: X
authors.initials: X
authors.lastname: Michalet
authors.forename: R A
authors.initials: RA
authors.lastname: Colyer
authors.forename: G
authors.initials: G
authors.lastname: Scalia
authors.forename: A
authors.initials: A
authors.lastname: Ingargiola
authors.forename: R
authors.initials: R
authors.lastname: Lin
authors.forename: J E
authors.initials: JE
authors.lastname: Millaud
authors.forename: S
authors.initials: S
authors.lastname: Weiss
authors.forename: Oswald H W
authors.initials: OH
authors.lastname: Siegmund
authors.forename: Anton S
authors.initials: AS
authors.lastname: Tremsin
authors.forename: John V
authors.initials: JV
authors.lastname: Vallerga
authors.forename: A
authors.initials: A
authors.lastname: Cheng
authors.forename: M
authors.initials: M
authors.lastname: Levi
authors.forename: D
authors.initials: D
authors.lastname: Aharoni
authors.forename: K
authors.initials: K
authors.lastname: Arisaka
authors.forename: F
authors.initials: F
authors.lastname: Villa
authors.forename: F
authors.initials: F
authors.lastname: Guerrieri
authors.forename: F
authors.initials: F
authors.lastname: Panzeri
authors.forename: I
authors.initials: I
authors.lastname: Rech
authors.forename: A
authors.initials: A
authors.lastname: Gulinatti
authors.forename: F
authors.initials: F
authors.lastname: Zappa
authors.forename: M
authors.initials: M
authors.lastname: Ghioni
authors.forename: S
authors.initials: S
authors.lastname: Cova
mesh_terms: D019295:Computational Biology; D004058:Diffusion; D004583:Electrons; D004867:Equipment Design; D005453:Fluorescence; D008856:Microscopy, Fluorescence / Q000379:methods*; D008968:Molecular Conformation; D057054:Molecular Imaging / Q000295:instrumentation* / Q000379:methods; D017785:Photons; D012680:Sensitivity and Specificity; D013997:Time Factors