Design study of a micro illumination platform based on GaN microLED arrays.

The design study of a micro illumination tool based on GaN microLED arrays is presented. The high spatio-temporal resolution and the capability of generating fully customized optical patterns that characterize the proposed platform would enable the manipulation of biological systems, e.g., for optogenetics applications. Based on ray tracing simulations, the design aspects that mainly affect the device performance have been identified, and the related structural parameters have been optimized to improve the extraction efficiency and the spatial resolution of the resulting light patterns. Assuming that the device is a bottom emitter, and the light is extracted from the n-side, the presence of mesa-structures on the p-side of the GaN layer can affect both the efficiency and the resolution, being optimized for different values of the mesa-side inclination angle. The full width at half maximum (FWHM) of the extracted spots is mainly determined by the substrate thickness, and the relation between the FWHM and the array pitch represents a criterion to define the resolution. Namely, when F W H M

A Compact Raster Lensless Microscope Based on a Microdisplay.

Lensless microscopy requires the simplest possible configuration, as it uses only a light source, the sample and an image sensor. The smallest practical microscope is demonstrated here. In contrast to standard lensless microscopy, the object is located near the lighting source. Raster optical microscopy is applied by using a single-pixel detector and a microdisplay. Maximum resolution relies on reduced LED size and the position of the sample respect the microdisplay. Contrarily to other sort of digital lensless holographic microscopes, light backpropagation is not required to reconstruct the images of the sample. In a mm-high microscope, resolutions down to 800 nm have been demonstrated even when measuring with detectors as large as 138 µm × 138 µm, with field of view given by the display size. Dedicated technology would shorten measuring time.

Pursuing the Diffraction Limit with Nano-LED Scanning Transmission Optical Microscopy.

Recent research into miniaturized illumination sources has prompted the development of alternative microscopy techniques. Although they are still being explored, emerging nano-light-emitting-diode (nano-LED) technologies show promise in approaching the optical resolution limit in a more feasible manner. This work presents the exploration of their capabilities with two different prototypes. In the first version, a resolution of less than 1 µm was shown thanks to a prototype based on an optically downscaled LED using an LED scanning transmission optical microscopy (STOM) technique. This research demonstrates how this technique can be used to improve STOM images by oversampling the acquisition. The second STOM-based microscope was fabricated with a 200 nm GaN LED. This demonstrates the possibilities for the miniaturization of on-chip-based microscopes.

Nano illumination microscopy: a technique based on scanning with an array of individually addressable nanoLEDs.

In lensless microscopy, spatial resolution is usually provided by the pixel density of current digital cameras, which are reaching a hard-to-surpass pixel size / resolution limit over 1 µm. As an alternative, the dependence of the resolving power can be moved from the detector to the light sources, offering a new kind of lensless microscopy setups. The use of continuously scaled-down Light-Emitting Diode (LED) arrays to scan the sample allows resolutions on order of the LED size, giving rise to compact and low-cost microscopes without mechanical scanners or optical accessories. In this paper, we present the operation principle of this new approach to lensless microscopy, with simulations that demonstrate the possibility to use it for super-resolution, as well as a first prototype. This proof-of-concept setup integrates an 8 × 8 array of LEDs, each 5 × 5 µm2 pixel size and 10 µm pitch, and an optical detector. We characterize the system using Electron-Beam Lithography (EBL) pattern. Our prototype validates the imaging principle and opens the way to improve resolution by further miniaturizing the light sources.

A Point-of-Care Device for Molecular Diagnosis Based on CMOS SPAD Detectors with Integrated Microfluidics.

We describe the integration of techniques and technologies to develop a Point-of-Care for molecular diagnosis PoC-MD, based on a fluorescence lifetime measurement. Our PoC-MD is a low-cost, simple, fast, and easy-to-use general-purpose platform, aimed at carrying out fast diagnostics test through label detection of a variety of biomarkers. It is based on a 1-D array of 10 ultra-sensitive Single-Photon Avalanche Diode (SPAD) detectors made in a 0.18 µm High-Voltage Complementary Metal Oxide Semiconductor (HV-CMOS) technology. A custom microfluidic polydimethylsiloxane cartridge to insert the sample is straightforwardly positioned on top of the SPAD array without any alignment procedure with the SPAD array. Moreover, the proximity between the sample and the gate-operated SPAD sensor makes unnecessary any lens or optical filters to detect the fluorescence for long lifetime fluorescent dyes, such as quantum dots. Additionally, the use of a low-cost laser diode as pulsed excitation source and a Field-Programmable Gate Array (FPGA) to implement the control and processing electronics, makes the device flexible and easy to adapt to the target label molecule by only changing the laser diode. Using this device, reliable and sensitive real-time proof-of-concept fluorescence lifetime measurement of quantum dot QdotTM 605 streptavidin conjugate is demonstrated.

Dynamic range extension of SiPM detectors with the time-gated operation.

The silicon photomultiplier (SiPM) is a novel detector technology that has undergone a fast development in the last few years, owing to its single-photon resolution and ultra-fast response time. However, the typical high dark count rates of the sensor may prevent the detection of low intensity radiation fluxes. In this article, the time-gated operation with short active periods in the nanosecond range is proposed as a solution to reduce the number of cells fired due to noise and thus increase the dynamic range. The technique is aimed at application fields that function under a trigger command, such as gated fluorescence lifetime imaging microscopy.

Fluorescence Multi-Detection Device Using a Lensless Matrix Addressable microLED Array.

A Point-of-Care system for molecular diagnosis (PoC-MD) is described, combining GaN and CMOS chips. The device is a micro-system for fluorescence measurements, capable of analyzing both intensity and lifetime. It consists of a hybrid micro-structure based on a 32 × 32 matrix addressable GaN microLED array, with square LEDs of 50 µm edge length and 100 µm pitch, with an underneath wire bonded custom chip integrating their drivers and placed face-to-face to an array of 16 × 16 single-photon avalanche diodes (SPADs) CMOS. This approach replaces instrumentation based on lasers, bulky optical components, and discrete electronics with a full hybrid micro-system, enabling measurements on 32 × 32 spots. The reported system is suitable for long lifetime (>10 ns) fluorophores with a limit of detection ~1/4 µM. Proof-of-concept measurements of streptavidin conjugate Qdot™ 605 and Amino PEG Qdot™ 705 are demonstrated, along with the device ability to detect both fluorophores in the same measurement.

A Novel Approach for a Chip-Sized Scanning Optical Microscope.

The recent advances in chip-size microscopy based on optical scanning with spatially resolved nano-illumination light sources are presented. This new straightforward technique takes advantage of the currently achieved miniaturization of LEDs in fully addressable arrays. These nano-LEDs are used to scan the sample with a resolution comparable to the LED sizes, giving rise to chip-sized scanning optical microscopes without mechanical parts or optical accessories. The operation principle and the potential of this new kind of microscope are analyzed through three different implementations of decreasing LED dimensions from 20 µm down to 200 nm.

Directly addressable GaN-based nano-LED arrays: fabrication and electro-optical characterization.

The rapid development of display technologies has raised interest in arrays of self-emitting, individually controlled light sources atthe microscale. Gallium nitride (GaN) micro-light-emitting diode (LED) technology meets this demand. However, the current technology is not suitable for the fabrication of arrays of submicron light sources that can be controlled individually. Our approach is based on nanoLED arrays that can directly address each array element and a self-pitch with dimensions below the wavelength of light. The design and fabrication processes are explained in detail and possess two geometries: a 6 × 6 array with 400 nm LEDs and a 2 × 32 line array with 200 nm LEDs. These nanoLEDs are developed as core elements of a novel on-chip super-resolution microscope. GaN technology, based on its physical properties, is an ideal platform for such nanoLEDs.

A Compact Analog Histogramming SPAD-Based CMOS Chip for Time-Resolved Fluorescence.

Time-resolved fluorescence measurement is extraordinarily powerful in the analysis of substances due to its effectiveness in eliminating measurement artifacts. Some fluorescence measurements are still conducted on CMOS chips with the decay times determined after reading the data off the chip and fitting the fluorescence decay histogram. We present a novel approach in which an analog CMOS chip divides the fluorescence decay time into slices and classifies the photons according to their arrival times at a CMOS SPAD sensor. The chip was fabricated in a 1P6M 0.18 µm HV-CMOS process. The slice timings can be tailored from 168 ps to 4.9 ns, covering most fluorescence decay times. 9 timing windows are generated per pixel that count up to 13 b each, with a resolution of 0.16 mV/photon, for a maximum output voltage of 1.3 V, in an area of 150 µm × 50 µm. Here, we report on the first practical application of this circuit, which integrates an array of 5 pixels in a single chip and has an excitation light and a microfluidic chip of up to 3 channels. This system could determine the decay time of quantum dots in 20 nl of solution. Thus, this paper could help in the development of a point-of-care device based on time-resolved fluorescence.