Miniature microscopy provides a transformative approach to observe objects and enable continuous monitoring with ultra-compact microscopes attached directly to specimens, facilitating parallel analysis. This innovation is particularly valuable for applications such as drug discovery using organ-on-a-chip devices, which require the assessment of numerous drug/sample pairs prior to clinical trials. Ultra-compact microscopes were previously limited to brightfield techniques, which prevented the use of powerful tools like fluorescent microscopy. In this work, we present a miniature microscope with integrated fluorescence measurement capabilities. This microscope consists of a custom chip with a 10 μm-diameter single-photon avalanche diode (SPAD) faced to a 640 × 480 InGaN/GaN 4 μm-pitch LED microdisplay. It operates in raster mode, activating individual LEDs to map specimens in 2D while measuring fluorescence light with the SPAD chip. Our results demonstrate its suitability for life sciences imaging. For example, we observed a muscle-ona-chip stained with Alexa Fluor 488 to study drug efficacy on sarcopenia. Furthermore, these microscopes exhibit superior speed compared to the previously reported ultra-compact brightfield microscopes, achieving a 240-fold increase in imaging rate by means of hardware controller integration on FPGA.
KEYWORDS: Sensors, Cameras, Microscopy, Light emitting diodes, Photodetectors, Signal to noise ratio, Time metrology, Statistical methods, Optical testing, Single photon
In modern single pixel microscopy techniques, like Nano-Illumination Microscopy, long measurement times can become a major issue, especially when imaging biological tissues with large field of view. Usually, light intensity measurements are performed with CMOS pixels, with typical integration times around tens of milliseconds. In this work, we propose to obtain a light intensity measurement indirectly by applying statistical techniques to the photon arrival times gathered with an SPAD photodetector. We will present how the different statistical measurements can be used to minimize the total acquisition time and minimize also the hardware required. In this work, with captures of 256 SPAD measurements, reducing measurement time from 50ms to 50us. The dynamic range is extended by combining multiple statistical techniques with standard intensity measurements. This paves the way to practical Nano-Illumination Microscopy and other single pixel microscopy techniques.
In this work we present a new microscope based on Nano-illumination microscopy (NIM), i.e., an innovative technique based on a 2D array of nano-Light-Emitting Diodes (LEDs) used to illuminate a sample. The key point of this method is that the pitch of the LED array fixes the spatial resolution. So, potentially, with LED pitches lower than the diffraction limit, super resolution could be achieved. While nanometer sized LEDs are not available yet, we present a prototype based on optical downscaling of a single 5µm lateral size LED. Extended Field-of-View (FOV) is obtained by mechanical movement with nanopositioners. Aspects of NIM microscopy such as its size, its flexibility in the sensing hardware or its potential for fluorescence, make it a perfect candidate to enhance emerging sensing applications in different fields, but especially life science (medical imaging, genomics, ...). We demonstrate the possibilities of the NIM technique with patterns as well as with biological samples.
Nano-Illumination Microscopy (NIM) is a technique that provides compact microscopes but at the present time only setups with limited Field-of-View (FOV) have been presented. Existing NIM setups reconstruct the image by measuring the light intensity that passes through the specimen when switching after one another the light emitting diodes (LEDs) on an array. The resolution of NIM is related to the LEDs pitch, while the FOV to the total area covered by the array. The first prototypes were demonstrated with 10 μm-pitch GaN-based 8x8 LED arrays giving rise to 80x80 μm2 FOV. This work presents the first electronically-activated Scanning Transmission Optical Microscope (eSTOM) built with an Organic LED-on-silicon micro-display with 5 μm LEDs pitch, providing a FOV of 3.6 × 1.28 mm2 . It is combined with a CMOS optical sensor with no other optical or mechanical components. We demonstrate how downscaling of the OLED array by means of optical lenses allows to further reduce the size of the light sources to explore the technique in more detail. Here we show steps towards the utility of NIM as a practical, low-cost and compact microscopy technique for biophotonics and many other applications.
This work presents a compact low-cost and straightforward shadow imaging microscopy technique based on spatially resolved nano-illumination instead of spatially resolved detection. Independently addressable nano-LEDs on a regular 2D array provide the resolution of the microscope by illuminating the sample in contact with the LED array and creating a shadow image in a photodetector located on the opposite side. The microscope prototype presented here is composed by a GaN chip with an 8x8 array of 5μm-LEDs with 10 μm pitch light sources and a commercial CMOS image sensor with integrated lens used as a light collector. We describe the microscope prototype and analyze the effect of the sensing area size on image reconstruction.
This work presents a first prototype for a new approach to microscopy: a system basing its resolving power on the light emitters instead of the sensors, without using lenses. This new approach builds on the possibility of making LEDs smaller than current technology sensors, offering a new approach to microscopy we plan on developing towards superresolution. The microscope consists on a SPAD based camera, a 8x8 LED array with 5x5 μm LEDs distributed with a pitch of 10 μm, and discrete driving electronics to control them. We present simulations of the system, as well as the first microscope prototype implementing the method, and the results obtained through it.
KEYWORDS: Sensors, Single photon, Avalanche photodiodes, CMOS technology, Electronics, Avalanche photodetectors, Signal to noise ratio, Electron beams, Diodes, TCAD
Advances in SPAD arrays propose improving the fill factor by confining several SPADs in the same well, with a main issue related to crosstalk. For measurements triggered only in well-defined time periods that can be known in advance, the pixels can be inhibited before the arrival of the crosstalk charge. This paper reports the crosstalk characterization of in an array of SPADs fabricated in a conventional CMOS technology in the same n-well (fill factor 67%). A long gating time gives a crosstalk not less than 2.75%, while reducing it below 2.5 ns completely eliminates crosstalk, as predicted by the theory and by TCAD simulations.
KEYWORDS: Avalanche photodetectors, Sensors, Electron beams, Monte Carlo methods, Single photon detectors, Scanning electron microscopy, Oxides, Particles, Photodetectors, Quenching (fluorescence)
Single-photon avalanche diodes (SPADs) are nowadays the most consolidate solid-state alternative to photomultiplier
tubes and time-correlated single-photon counting. Optical benches are used for the characterization of the noise figures
of these detectors, including dark count, afterpulsing effects and cross-talk. With accurate optical setups it is possible to
obtain resolutions down to 5 microns, but with today's technologies, this spot size can cover more than one single pixel.
Moreover, on other common and envisaged applications like particle detection in Nuclear and High Energy Physics or as
silicon photomultipliers for Cerenkov telescopes, this does not allow to observe what happens when a charge is
generated between consecutive pixels.
This work presents the innovative characterization of single-photon detectors with the aid of the electron beam generated
in a dual beam FIB/SEM apparatus. A simple setup allows a very good control of the dose and the spot down to 5 nm at
30 keV, The characterization has been proven in photodetectors fabricated in a standard CMOS technology. The results
have been validated by comparison with those obtained by optical setups, with simulation with PENELOPE (Penetration
and Energy Loss of Positrons and Electrons) and by technology simulations with ISE-tCAD.
In the last years the fabrication of SPAD cameras has become one of the main fields of interest in 3-D imaging and bioapplications.
In this paper we present the comparison between two standard CMOS technologies to fabricate SPADs
cameras. The two technologies used in the comparison are a high voltage 0.35μm technology from AMS and a high
integration 130nm technology from STM. The advantage of using a standard CMOS technology among a dedicated is
the possibility of integrating the control/reading electronics into the same die. Neither of the processes is optimized for
optical applications, and no post-processing has been applied to improve the features.
The technologies have been selected due to the different integration density, and different intrinsic process parameters
with similar cost. Comparison has been done by fabricating several structures in both technologies which allow
analyzing sensibility, noise, and time response.
Experimental results show that the high voltage technology has a lower level of dark counts than the 130nm. Instead, the
high integration technology has a shorter quenching time, 1.5ns, which reduces the afterpulsing events to a negligible
level. In optical applications it is important to have a high integration of the camera reducing the pitch of the pixel, while
noise effects can be corrected in post-processing. For low frequency events, such as high energetic particle tracking, the
noise frequency has to be lower, but it is also required a high fill factor. Depending on the specific application this
analysis allows to opt for the most suitable technology.
It is described the architecture of the electronics for the control of a wireless endoscopic capsule with locomotive capabilities and advanced sensing and actuating functions. Special emphasis is done to the description of the driver used for locomotion, which is the most innovative element in the capsule.
KEYWORDS: System on a chip, Virtual colonoscopy, Solar cells, Telecommunications, Computer programming, Prototyping, Sensors, Clocks, Control systems, Optical communications
This paper presents a System On Chip (SoC) designed specifically to control a mm3- sized microrobot called I-SWARM. The robot is intended to be part of a colony of 1000 I-SWARM robots for studying swarm behavior in real time and in a real swarm. The SoC offers a well-suited hardware platform to run multi-agent systems software. It is composed of an 8051 microcontroller with 2 kB of data memory and 8 kB of program memory. The processor is provided with specific hardware modules for controlling the locomotion unit, the communications and the vibrating contact sensor of the robot. These modules perform basic tasks as movements or communications so the 8051 can focus on processing data and taking decisions. With these capabilities, the robot is able to avoiding collisions with other members of the swarm, performing cooperative tasks, sharing information and executing specialized tasks. The SoC has been fabricated with a 0.13 &mgr;m ultra low power CMOS process of STMicroelectronics and consumes less than 1 mW.
Nowadays Atomic Force Microscopy is one of the most extended techniques performed in biological measurements. Due to the higher flexibility in respect to conventional equipments, a novel approach in this field is the use of a microrobot equipped with an AFM tool. In this paper it is presented an integrated controller for an AFM tool assembled in a 1 cm3 wireless microrobot. The AFM tool is mounted on the tip of a rotational piezoelectric actuator arm. It consists on a XYZ positioning scanner, based in 4 piezoelectric stacked actuators, and an AFM piezoresistance probe. Two types of AFM working modes are implemented in the controller, i.e., nanoidentation and AFM scanning. Correction of the mismatch of the piezoactuators composing the arm is possible. A programmable PID control is included in the controller in order to get more flexibility in terms of scanning speed and resolution. An IrDA protocol is used to program the parameters of the AFM tool controller and the positioning of the robot in the working area. Then the values of the nanoindentation or of the scanning can be read through the IrDA interface without any other external action.
Due to the strong power and area restrictions, the controller has been implemented in specific logic in a 0.35um technology. The design has been done using functional specifications with high level tools and RTL synthesis. The AFM scanner can be positioned with a resolution of 10 nm and scan areas up to 1 μm2 with an expected vertical resolution of 1nm.
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