Ivan Michel Antolovic

Nonuniformity Analysis of a 65-kpixel CMOS SPAD Imager

While CMOS single-photon avalanche diode (SPAD) technology has steadily advanced, improving noise, timing resolution, and sensitivity, spatial resolution has been increasing as well. The increase in the number of pixels has made a comprehensive analysis of nonuniformity and its effects meaningful, allowing a more accurate comparison of SPAD imagers with other high-end scientific imagers, such as electron multiplying charge-coupled device and scientific CMOS. A comprehensive nonuniformity analysis was conducted on a 512 × 128 pixel gated SPAD imager, where dark noise, afterpulsing, crosstalk, signal response, and shot noise were measured. This analysis has led to a variety of postprocessing algorithms to improve the linearity of the response as for example required by ground state depletion microscopy-based superresolution microscopy and other techniques. We derived a new correction formula for the count rate applicable to 1-b SPAD imagers, and we measured a significant improvement of photon detection efficiency using microlenses. These techniques were used to validate the suitability of the imager in fluorescence microscopy examples.

Imaging free and bound NADH towards cancer tissue detection using FLIM system based on SPAD array

Single photon detectors allow us to work with the weakest signals such as auto-fluorescent biological sources. In combination with time gated operation mode, an array of detectors can be used as Fluorescence Lifetime Imaging system with extremely high sensitivity.

Fluorescence lifetime imaging using a single photon avalanche diode array sensor (Conference Presentation)

Single photon detectors allows us work with the weakest fluorescence signals. Single photon arrays, combined with ps-controlled gating allow us to create image maps of fluorescence lifetimes, which can be used for in-vivo discrimination of tissue activity. Here we present fluorescence lifetime imaging using the ‘SwissSPAD’ sensor, a 512-by-128-pixel array of gated single photon detectors, fabricated in a standard high-voltage 0.35 μm CMOS process. We present a protocol for spatially resolved lifetime measurements where the lifetime can be retrieved for each pixel. We demonstrate the system by imaging patterns of Fluorescein and Rhodamine B on test slides, as well as measuring mixed samples to retrieve both components of the decay lifetime. The single photon sensitivity of the sensor creates a valuable instrument to perform live cell or live animal (in vivo) measurements of the weak autofluorescent signals, for example distinguishing unlabelled free and bound NADH. Our ultimate goal is to create a real time fluorescence lifetime imaging system, possibly integrated into augmented reality goggles, which could allow immediate discrimination of in vivo tissues.