High-throughput single-molecule fluorescence spectroscopy using parallel detection.

Solution-based single-molecule fluorescence spectroscopy is a powerful new experimental approach with applications in all fields of natural sciences. The basic concept of this technique is to excite and collect light from a very small volume (typically femtoliter) and work in a concentration regime resulting in rare burst-like events corresponding to the transit of a single-molecule. Those events are accumulated over time to achieve proper statistical accuracy. Therefore the advantage of extreme sensitivity is somewhat counterbalanced by a very long acquisition time. One way to speed up data acquisition is parallelization. Here we will discuss a general approach to address this issue, using a multispot excitation and detection geometry that can accommodate different types of novel highly-parallel detector arrays. We will illustrate the potential of this approach with fluorescence correlation spectroscopy (FCS) and single-molecule fluorescence measurements obtained with different novel multipixel single-photon counting detectors.

Monolithic time to amplitude converter for time correlated single photon counting.

Time correlated single photon counting techniques allow in depth examinations of very important chemical and biological processes involving complex interactions of biomolecules. Understanding of these processes is of the utmost importance to address vital medical issues such as the origin and growth of tumors. Modern developments of fluorescence analysis techniques require compact, low cost, and high performance instrumentation, and a major path to these goals is the successful integration of the electronics. In this paper we present a fully monolithic time to amplitude converter, built in standard 0.35 microm CMOS technology, characterized by good time resolution (60 ps), low differential nonlinearity (better than 0.5% rms), short dead time (80 ns), low power dissipation (60 mW), and low area occupation (1.8x1.4 mm(2)).

High-linearity analog-to-digital acquisition board for photon-timing applications.

Nowadays a wide range of scientific applications require the detection of very weak and fast luminescence signals in the time domain. Single molecule spectroscopy, diffuse optical tomography, time-resolved emission spectra are only a few examples among them. Advanced time-correlated single photon counting, that relies on multidetector and multidimensional acquisition, reveals itself as a powerful technique to gather deeper insight into various photochemical and biological processes. In this paper we present a high-speed, high-linearity A/D acquisition board that is designed as a building block for a compact, low-cost, multidetector acquisition system for photon-timing experiments. The prototype has been experimentally tested and the results fully comply the goal design specifications.

Self-suppression of reset induced triggering in picosecond SPAD timing circuits.

We present a new photon timing circuit that achieves a time resolution of 35 ps full width at half maximum with single photon avalanche diodes having active area diameters up to 200 microm. The timing circuit is based on a double avalanche current sensing network that makes it particularly suited to operation at high photon counting rates. Thanks to its self-adjusting capabilities, no trimming is needed even when changing the photodetector operating conditions over a wide range.

Monolithic active quenching and picosecond timing circuit suitable for large-area single-photon avalanche diodes.

A new integrated active quenching circuit (i-AQC) designed in a standard CMOS process is presented, capable of operating with any available single photon avalanche diode (SPAD) over wide temperature range. The circuit is suitable for attaining high photon timing resolution also with wide-area SPADs. The new i-AQC integrates the basic active-quenching loop, a patented low-side timing circuit comprising a fast pulse pick-up scheme that substantially improves time-jitter performance, and a novel active-load passive quenching mechanism (consisting of a current mirror rather than a traditional high-value resistor) greatly improves the maximum counting rate. The circuit is also suitable for portable instruments, miniaturized detector modules and SPAD-array detectors. The overall features of the circuit may open the way to new developments in diversified applications of time-correlated photon counting in life sciences and material sciences.