Multiplexed single-photon counting. I. A time-correlated fluorescence lifetime camera

Abstract

Photoluminescence lifetime microscope spectrometer based on timecorrelated singlephoton counting with an avalanche diode detector Rev. Sci. Instrum. 61, 11 (1990); 10.1063/1.1141885Time response of small sideon photomultiplier tubes in timecorrelated singlephoton counting measurements Rev. Sci. Instrum. 59, 499 (1988);We report the operation of a new type of fluorescence lifetime imaging camera based on the time-correlated single-photon counting ͑TCSPC͒ technique. To the best of our knowledge the application-specific integrated circuit ͑ASIC͒ used in the camera is the first ASIC designed for the field of fluorescence. The ASIC also forms the basis for the first read-out system for single-photon timing array detectors and is capable of multiplexing and routing counts from up to sixteen detection channels, while preserving their timing characteristics with picosecond resolution. In conjunction with an array detector such as a multianode MCP-PM this ASIC allows multiple fluorescence decays to be routinely and simultaneously acquired using a single set of TCSPC timing electronics. To demonstrate one practical application of this technology, we have observed for the first time the spatial distribution of fluorescence lifetime contours through a strongly self-absorbing sample, and the effects observed demonstrate how differences in optical geometry can contribute to the lack of consistency between results obtained in different laboratories.

Fluorescence lifetime imaging in scanning microscopes: acquisition speed, photon economy and lifetime resolution

“…This limits the shortest lifetimes that can be measured with this type of tube to similar values. In order to measure shorter lifetimes, micro channel plate PMTs (MCP‐PMTs) (McLoskey et al ., 1996) can be used. MCP‐PMTs have quantum efficiencies in the order of 10% and exhibit transit time spreads as low as 25 ps, which makes them particularly suited for the measurement of fast decays.…”

Fluorescence-Lifetime Imaging Techniques for Microscopy

“…It uses a high-speed CCD camera modified to collect lifetime data. The heterodyning technique coupled with a fast camera yields several unique advantages over previously reported camera systems (Lakowicz and Berndt, 1991;Wang et al, 1991;Gadella et al, 1993;Oida et al, 1993;Morgan et al, 1995;McLoskey et al, 1996). First, this instrument is capable of generating lifetime images in a few seconds.…”

“…The basic approaches are to use a gated image intensifier coupled to a camera or a position-sensitive photomultiplier. Both frequency-domain (Wang et al, 1989;Marriott et al, 1991;Lakowicz and Berndt, 1991;So et al, 1994;Morgan et al, 1995) and time-domain (Wang et al, 1991;Oida et al, 1993;McLoskey et al, 1996) techniques have been developed.…”

Wide-field TCSPC: methods and applications

“…Advanced read-out architectures that are able to resolve multi-photon hits have been designed [123], however they do not fundamentally solve the problem that the count rate is limited by the position read-out electronics rather than the need to avoid overlapping events. [40,124,125] and pixelated read-out anodes allow the positions of the events from the detector to be read out directly at each pixel. An 8 8 × pixel anode read-out with 43 ps time resolution that uses application specific integrated circuits (ASICs) originally developed in CERN has been demonstrated, and a 32 32 × pixel detector has been planned, but the large pixel size of 0.5 mm and limited number of pixels mean that the development of these detectors is aimed more at highcontent screening applications [126].…”