Field-portable pixel super-resolution colour microscope.

Based on partially-coherent digital in-line holography, we report a field-portable microscope that can render lensfree colour images over a wide field-of-view of e.g., >20 mm(2). This computational holographic microscope weighs less than 145 grams with dimensions smaller than 17×6×5 cm, making it especially suitable for field settings and point-of-care use. In this lensfree imaging design, we merged a colorization algorithm with a source shifting based multi-height pixel super-resolution technique to mitigate 'rainbow' like colour artefacts that are typical in holographic imaging. This image processing scheme is based on transforming the colour components of an RGB image into YUV colour space, which separates colour information from brightness component of an image. The resolution of our super-resolution colour microscope was characterized using a USAF test chart to confirm sub-micron spatial resolution, even for reconstructions that employ multi-height phase recovery to handle dense and connected objects. To further demonstrate the performance of this colour microscope Papanicolaou (Pap) smears were also successfully imaged. This field-portable and wide-field computational colour microscope could be useful for tele-medicine applications in resource poor settings

Development of a Portable CMOS Time-Domain Fluorescence Lifetime Imager

Modern laboratory equipments to measure the excited-state lifetime of fluorophores usually include an expensive picosecond pulsed-laser excitation source, a fragile photomultiplier tube, and a large instrument body for optics. A portable and robust device to make fluorescence lifetime measurement in nanosecond scale is of great attraction for chemists and biologists. This dissertation reports the development of a portable LED time-domain fluorimeter from an all-solid-state discrete-component prototype to its advanced CMOS integrated circuit implementation. The motivation of the research is to develop a multiplexed fluorimeter for point-of-care diagnosis. Instruments developed by this novel method have higher fill factor, are more portable, and are fabricated at lower cost

High-Speed Wide-Field Time-Correlated Single-Photon Counting Fluorescence Lifetime Imaging Microscopy

Fluorescence microscopy is a powerful imaging technique used in the biological sciences to identify labeled components of a sample with specificity. This is usually accomplished through labeling with fluorescent dyes, isolating these dyes by their spectral signatures with optical filters, and recording the intensity of the fluorescent response. Although these techniques are widely used, fluorescence intensity images can be negatively affected by a variety of factors that impact the fluorescence intensity. Fluorescence lifetime imaging microscopy (FLIM) is an imaging technique that is relatively immune to intensity fluctuations and also provides the unique ability to directly monitor the microenvironment surrounding a fluorophore. Despite the benefits associated with FLIM, the applications to which it is applied are fairly limited due to long image acquisition times and high cost of traditional hardware. Recent advances in complementary metal-oxide-semiconductor (CMOS) single-photon avalanche diodes (SPADs) have enabled the design of low-cost imaging arrays that are capable of recording lifetime images with acquisition times greater than one order of magnitude faster than existing systems. However, these SPAD arrays have yet to realize the full potential of the technology due to limitations in their ability to handle the vast amount of data generated during the commonly used time-correlated single-photon counting (TCSPC) lifetime imaging technique. This thesis presents the design, implementation, characterization, and demonstration of a high speed FLIM imaging system. The components of this design include a CMOS imager chip in a standard 0.13 μm technology containing a custom CMOS SPAD, a 64-by-64 array of these SPADs, pixel control circuitry, independent time-to-digital converters (TDCs), a FLIM specific datapath, and high bandwidth output buffers. In addition to the CMOS imaging array, a complete system was designed and implemented using a printed circuit board (PCB) for capturing data from the imager, creating histograms for the photon arrival data using field-programmable gate arrays, and transferring the data to a computer using a cabled PCIe interface. Finally, software is used to communicate between the imaging system and a computer.The dark count rate of the SPAD was measured to be only 231 Hz at room temperature while maintaining a photon detection probability of up to 30\%. TDCs included on the array have a 62.5 ps resolution and a 64 ns range, which is suitable for measuring the lifetime of most biological fluorophores. Additionally, the on-chip datapath was designed to handle continuous data transfers at rates capable of supporting TCSPC-based lifetime imaging at 100 frames per second. The system level implementation also provides sufficient data throughput for transferring up to 750 frames per second from the imaging system to a computer. The lifetime imaging system was characterized using standard techniques for evaluating SPAD performance and an electrical delay signal for measuring the TDC performance. This thesis concludes with a demonstration of TCSPC-FLIM imaging at 100 frames per second -- the fastest 64-by-64 TCSPC FLIM that has been demonstrated. This system overcomes some of the limitations of existing FLIM systems and has the potential to enable new application domains in dynamic FLIM imaging

SPAD Figures of Merit for Photon-Counting, Photon-Timing, and Imaging Applications: A Review

Single-photon avalanche diodes (SPADs) emerged as the most suitable photodetectors for both single-photon counting and photon-timing applications. Different complementary metal-oxide-semiconductor (CMOS) devices have been reported in the literature, with quite different performance and some excelling in just few of them, but often at different operating conditions. In order to provide proper criteria for performance assessment, we present some figures of merit (FoMs) able to summarize the typical SPAD performance (i.e., photon detection efficiency, dark counting rate, afterpulsing probability, hold-off time, and timing jitter) and to identify a proper metric for SPAD comparisons, when used either as single-pixel detectors or in imaging arrays. The ultimate goal is not to define a ranking list of best-in-class detectors, but to quantitatively help the end-user to state the overall performance of different SPADs in either photon-counting, timing, or imaging applications. We review many CMOS SPADs from different research groups and companies, we compute the proposed FoMs for all them and, eventually, we provide an insight on present CMOS SPAD technologies and future trends

Advanced photon counting techniques for long-range depth imaging

The Time-Correlated Single-Photon Counting (TCSPC) technique has emerged as a candidate approach for Light Detection and Ranging (LiDAR) and active depth imaging applications. The work of this Thesis concentrates on the development and investigation of functional TCSPC-based long-range scanning time-of-flight (TOF) depth imaging systems. Although these systems have several different configurations and functions, all can facilitate depth profiling of remote targets at low light levels and with good surface-to-surface depth resolution. Firstly, a Superconducting Nanowire Single-Photon Detector (SNSPD) and an InGaAs/InP Single-Photon Avalanche Diode (SPAD) module were employed for developing kilometre-range TOF depth imaging systems at wavelengths of ~1550 nm. Secondly, a TOF depth imaging system at a wavelength of 817 nm that incorporated a Complementary Metal-Oxide-Semiconductor (CMOS) 32×32 Si-SPAD detector array was developed. This system was used with structured illumination to examine the potential for covert, eye-safe and high-speed depth imaging. In order to improve the light coupling efficiency onto the detectors, the arrayed CMOS Si-SPAD detector chips were integrated with microlens arrays using flip-chip bonding technology. This approach led to the improvement in the fill factor by up to a factor of 15. Thirdly, a multispectral TCSPC-based full-waveform LiDAR system was developed using a tunable broadband pulsed supercontinuum laser source which can provide simultaneous multispectral illumination, at wavelengths of 531, 570, 670 and ~780 nm. The investigated multispectral reflectance data on a tree was used to provide the determination of physiological parameters as a function of the tree depth profile relating to biomass and foliage photosynthetic efficiency. Fourthly, depth images were estimated using spatial correlation techniques in order to reduce the aggregate number of photon required for depth reconstruction with low error. A depth imaging system was characterised and re-configured to reduce the effects of scintillation due to atmospheric turbulence. In addition, depth images were analysed in terms of spatial and depth resolution