[Direct Positron Emission Imaging Using Ultrafast Timing Performance Detectors].

This is an explanatory paper on Sun Il Kwon et al., Nat. Photon. 15: 914-918, 2021 and some parts of this manuscript are translated from the paper. Medical imaging modalities such as X-ray computed tomography, Magnetic resonance imaging, positron emission tomography (PET), and single photon emission computed tomography, require image reconstruction processes, consequently constraining them to form cylindrical shapes. However, among them, only PET can use additional information, so called time of flight, on an event-by-event basis. If coincidence time resolution (CTR) of PET detectors improved to 30 ps, which corresponds to spatial resolution of 4.5 mm, directly localizing electron-positron annihilation point is possible, allowing us to circumvent image reconstruction processes and free us from the geometric constraint. We call this concept direct positron emission imaging (dPEI). We have developed ultrafast radiation detectors by focusing on Cherenkov photon detection. Furthermore, the CTR of 32 ps being equivalent to 4.8 mm spatial resolution is achieved by combining deep learning-based signal processing with the detectors. In this article, we explain how we developed the detectors and demonstrated the first dPEI using different types of phantoms, how we will tackle limitations to be addressed to make the dPEI more practical, and how dPEI will emerge as an imaging modality in nuclear medicine.

Estimation of the Spatial Extent of the Transient Gain Drop in a Microchannel Plate.

The gain of the microchannel plate temporally drops after an ion initiates an electron avalanche. Electron multiplication was expected to deplete the charge from the microchannel wall and produce the depleted charge (wall charge). Moreover, it was reported that the gain drop occurred not only in the activated channels, where the electrons are multiplied, but also in the surrounding channels. One mechanism of the gain-drop spatial extension has been considered as that the wall charges in the activated channels change the electric field in the surrounding channels. Anacker et al. assumed that the wall charge is a uniform line charge; the gain-drop spatial extent should be proportional to the amount of the wall charges. We considered that the wall charges exponentially increased in the channel toward the exit. In this study, the electric field produced by the wall charges was calculated, considering the distribution of the wall charges. The transverse electric field generated by the wall charges was expected to disturb the electron trajectory near the channel exit and decrease the number of secondary electrons emitted per collision (gain per collision), resulting in a gain drop. The gain per collision was calculated to decrease by 22% for the position where the gain decreased significantly in the presence of the transverse electric field of 3×105 V/m. In our model, the gain-drop spatial extent extended proportionally to the square root of the wall charges when the distance from the activated channel exceeded 50 µm.

Characterization of microchannel plate detector response for the detection of native multiply charged high mass single ions in orthogonal-time-of-flight mass spectrometry using a Timepix detector.

Time-of-flight (TOF) systems are one of the most widely used mass analyzers in native mass spectrometry (nMS) for the analysis of non-covalent multiply charged bio-macromolecular assemblies (MMAs). Typically, microchannel plates (MCPs) are employed for high mass native ion detection in TOF MS. MCPs are well known for their reduced detection efficiency when impinged by large slow moving ions. Here, a position- and time-sensitive Timepix (TPX) detector has been added to the back of a dual MCP stack to study the key factors that affect MCP performance for MMA ions generated by nMS. The footprint size of the secondary electron cloud generated by the MCP on the TPX for each individual ion event is analyzed as a measure of MCP performance at each mass-to-charge (m/z) value and resulted in a Poisson distribution. This allowed us to investigate the dependency of ion mass, ion charge, ion velocity, acceleration voltage, and MCP bias voltage on MCP response in the high mass low velocity regime. The study of measurement ranges; ion mass = 195 to 802,000 Da, ion velocity = 8.4 to 67.4 km/s, and ion charge = 1+ to 72+, extended the previously examined mass range and characterized MCP performance for multiply charged species. We derived a MCP performance equation based on two independent ion properties, ion mass and charge, from these results, which enables rapid MCP tuning for single MMA ion detection.

Synchrotron radiation transmission by two coupled flat microchannel plates: new opportunities to control the focal spot characteristics.

An improved theoretical model to calculate the focal spot properties of coherent synchrotron radiation (SR) soft X-ray beams by combining and aligning two microchannel plates (MCPs) is presented. The diffraction patterns of the radiation behind the MCP system are simulated in the framework of the electrodynamical model of the radiation emission from two-dimensional finite antenna arrays. Simulations show that this particular optical device focuses the soft X-ray radiation in a circular central spot with a radius of ∼4 µm. The study points out that such MCP-based devices may achieve micrometre and sub-micrometre spot sizes as required by many applications in the soft X-ray range. Finally, based on experimental and theoretical results of the radiation transmission by this MCP-based device, a new method to characterize the spatial properties of brilliant SR sources is discussed.

Wave propagation and focusing of soft X-rays by spherical bent microchannel plates.

Synchrotron radiation sources have been used to study the focusing properties and angular distribution of X-ray radiation at the exit of spherically bent microchannel plates (MCPs). In this contribution it is shown how soft X-ray radiation at energies up to 1.5 keV can be focused by spherically bent MCPs with curvature radii R of 30 mm and 50 mm. For these devices, a focus spot is detectable at a distance between the detector and the MCP of less than R/2, with a maximum focusing efficiency up to 23% of the flux illuminating the MCP. The soft X-ray radiation collected at the exit of microchannels of spherically bent MCPs are analyzed in the framework of a wave approximation. A theoretical model for the wave propagation of radiation through MCPs has been successfully introduced to explain the experimental results. Experimental data and simulations of propagating radiation represent a clear confirmation of the wave channeling phenomenon for the radiation in spherically bent MCPs.

Evaluation of microchannel plate gain drops caused by high ion fluxes in time-of-flight mass spectrometry: A novel evaluation method using a multi-turn time-of-flight mass spectrometer.

Use of a two-stage microchannel plate (MCP) detector is common in time-of-flight (TOF) mass spectrometry because it shows excellent time resolution with sufficient gains. However, the gain drops significantly when the detector detects intense ion fluxes, such as matrix ions, by matrix-assisted laser desorption/ionization mass spectrometry. As a result, significant ion signals corresponding to analytes, such as proteins, are hidden, thereby hampering the mass spectral interpretation. However, details of this phenomenon have not previously been investigated using ions because of the lack of suitable measurement methods and apparatus. Thus, we herein report a novel method for controlling the TOF of two selected ions, as a function of time differences between each other using a multi-turn TOF mass spectrometer. This method involves the use of an isotope cluster of ions that fly in a figure-of-eight orbit and the extraction of an ion at a given lap number. A series of time differences (∆t) between two ions in a TOF spectrum can be generated using this method. We evaluated the time constants of gain recovery after high ion-flux detection for two sets of two-stage MCP detectors to obtain values of 1,600 and 180 µs for channel plate resistances of 500 and 71 MΩ, respectively. The obtained time constants from the gains determined at various values of ∆t were 0.48 and 0.38 fold (for 500 and 71 MΩ, respectively) of the values suggested from the channel plate resistance and capacitance.

Photon Counting Imaging with Low Noise and a Wide Dynamic Range for Aurora Observations.

The radiation intensity of observed auroras in the far-ultraviolet (FUV) band varies dramatically with location for aerospace applications, requiring a photon counting imaging apparatus with a wide dynamic range. However, combining high spatial resolution imaging with high event rates is technically challenging. We developed an FUV photon counting imaging system for aurora observation. Our system mainly consists of a microchannel plate (MCP) stack readout using a wedge strip anode (WSA) with charge induction and high-speed electronics, such as a charge sensitive amplifier (CSA) and pulse shaper. Moreover, we constructed an anode readout model and a time response model for readout circuits to investigate the counting error in high counting rate applications. This system supports global rates of 500 kilo counts, 0.610 dark counts s-1 cm-2 at an ambient temperature of 300 K and 111 µm spatial resolution at 400 kilo counts s-1 (kcps). We demonstrate an obvious photon count loss at incident intensities close to the counting capacity of the system. To preserve image quality, the response time should be improved and some noise performance may be sacrificed. Finally, we also describe the correlation between counting rate and imaging resolution, which further guides the design of space observation instruments.

Lightsheet fluorescence lifetime imaging microscopy with wide-field time-correlated single photon counting.

We report on wide-field time-correlated single photon counting (TCSPC)-based fluorescence lifetime imaging microscopy (FLIM) with lightsheet illumination. A pulsed diode laser is used for excitation, and a crossed delay line anode image intensifier, effectively a single-photon sensitive camera, is used to record the position and arrival time of the photons with picosecond time resolution, combining low illumination intensity of microwatts with wide-field data collection. We pair this detector with the lightsheet illumination technique, and apply it to 3D FLIM imaging of dye gradients in human cancer cell spheroids, and C. elegans.

MCP-based detectors: calibration and first photon radiation measurements.

Detectors based on microchannel plates (MCPs) are used to detect radiation from free-electron lasers. Three MCP detectors have been developed by JINR for the European XFEL (SASE1, SASE2 and SASE3 lines). These detectors are designed to operate in a wide dynamic range from the level of spontaneous emission to the SASE saturation level (between a few nJ up to 25 mJ), in a wide wavelength range from 0.05 nm to 0.4 nm for SASE1 and SASE2, and from 0.4 nm to 4.43 nm for SASE3. The detectors measure photon pulse energies with an anode and a photodiode. The photon beam image is observed with an MCP imager with a phosphor screen. At present, the SASE1 and SASE3 MCP detectors are commissioned with XFEL beams. Calibration and first measurements of photon radiation in multibunch mode are performed with the SASE1 and SASE3 MCPs. The MCP detector for SASE2 and its electronics are installed in the XFEL tunnel, technically commissioned, and are now ready for acceptance tests with the X-ray beam.

High-Sensitivity and Long-Life Microchannel Plate Processed by Atomic Layer Deposition.

As a key component of electron multiplier device, a microchannel plate (MCP) can be applied in many scientific fields. Pure aluminum oxide (Al2O3) as secondary electron emission (SEE) layer were deposited in the pores of MCP via atomic layer deposition (ALD) to overcome problems such as high dark current and low lifetime which often occur on traditional MCP. In this paper, we systematically investigate the morphology, element distribution, and structure of samples by scanning electron microscopy (SEM) and energy disperse spectroscopy (EDS), respectively. Output current of different thickness of Al2O3 was studied and an optimal thickness was found. Experimental tests show that the average gain of ALD-MCP was nearly five times better than that of traditional MCP, and the ALD-MCP showed better sensitivity and longer lifetime.

Reduction Temperature-Dependent Nanoscale Morphological Transformation and Electrical Conductivity of Silicate Glass Microchannel Plate.

Lead silicate glasses are fundamental materials to a microchannel plate (MCP), which is a two dimensional array of a microscopic channel charge particle multiplier. Hydrogen reduction is the core stage to determine the electrical conductivity of lead silicate glass MCP multipliers. The nanoscale morphologies and microscopic potential distributions of silicate glass at different reduction temperatures were investigated via atomic force microscope (AFM) and Kelvin force microscopy (KFM). We found that the bulk resistance of MCPs ballooned exponentially with the spacing of conducting islands. Moreover, bulk resistance and the spacing of conducting islands both have the BiDoseResp trend dependence on the hydrogen reduction temperature. Elements composition and valence states of lead silicate glass were characterized by X-ray photoelectron spectroscopy (XPS). The results indicated that the conducting island was an assemblage of the Pb atom originated from the reduction of Pb2+ and Pb4+. Thus, this showed the important influence of the hydrogen temperature and nanoscale morphological transformation on modulating the physical effects of MCPs, and opened up new possibilities to characterize the nanoscale electronic performance of multiphase silicate glass.

Microfluidic co-culture of pancreatic tumor spheroids with stellate cells as a novel 3D model for investigation of stroma-mediated cell motility and drug resistance.

BACKGROUND: Pancreatic stellate cells (PSCs), a major component of the tumor microenvironment in pancreatic cancer, play roles in cancer progression as well as drug resistance. Culturing various cells in microfluidic (microchannel) devices has proven to be a useful in studying cellular interactions and drug sensitivity. Here we present a microchannel plate-based co-culture model that integrates tumor spheroids with PSCs in a three-dimensional (3D) collagen matrix to mimic the tumor microenvironment in vivo by recapitulating epithelial-mesenchymal transition and chemoresistance. METHODS: A 7-channel microchannel plate was prepared using poly-dimethylsiloxane (PDMS) via soft lithography. PANC-1, a human pancreatic cancer cell line, and PSCs, each within a designated channel of the microchannel plate, were cultured embedded in type I collagen. Expression of EMT-related markers and factors was analyzed using immunofluorescent staining or Proteome analysis. Changes in viability following exposure to gemcitabine and paclitaxel were measured using Live/Dead assay. RESULTS: PANC-1 cells formed 3D tumor spheroids within 5 days and the number of spheroids increased when co-cultured with PSCs. Culture conditions were optimized for PANC-1 cells and PSCs, and their appropriate interaction was confirmed by reciprocal activation shown as increased cell motility. PSCs under co-culture showed an increased expression of α-SMA. Expression of EMT-related markers, such as vimentin and TGF-ß, was higher in co-cultured PANC-1 spheroids compared to that in mono-cultured spheroids; as was the expression of many other EMT-related factors including TIMP1 and IL-8. Following gemcitabine exposure, no significant changes in survival were observed. When paclitaxel was combined with gemcitabine, a growth inhibitory advantage was prominent in tumor spheroids, which was accompanied by significant cytotoxicity in PSCs. CONCLUSIONS: We demonstrated that cancer cells grown as tumor spheroids in a 3D collagen matrix and PSCs co-cultured in sub-millimeter proximity participate in mutual interactions that induce EMT and drug resistance in a microchannel plate. Microfluidic co-culture of pancreatic tumor spheroids with PSCs may serve as a useful model for studying EMT and drug resistance in a clinically relevant manner.

Band Offset Measurements in Atomic-Layer-Deposited Al2O3/Zn0.8Al0.2O Heterojunction Studied by X-ray Photoelectron Spectroscopy.

Pure aluminum oxide (Al2O3) and zinc aluminum oxide (Zn x Al1-x O) thin films were deposited by atomic layer deposition (ALD). The microstructure and optical band gaps (E g ) of the Zn x Al1-x O (0.2 ≤ x ≤ 1) films were studied by X-ray diffractometer and Tauc method. The band offsets and alignment of atomic-layer-deposited Al2O3/Zn0.8Al0.2O heterojunction were investigated in detail using charge-corrected X-ray photoelectron spectroscopy. In this work, different methodologies were adopted to recover the actual position of the core levels in insulator materials which were easily affected by differential charging phenomena. Valence band offset (ΔE V) and conduction band offset (ΔE C) for the interface of the Al2O3/Zn0.8Al0.2O heterojunction have been constructed. An accurate value of ΔE V = 0.82 ± 0.12 eV was obtained from various combinations of core levels of heterojunction with varied Al2O3 thickness. Given the experimental E g of 6.8 eV for Al2O3 and 5.29 eV for Zn0.8Al0.2O, a type-I heterojunction with a ΔE C of 0.69 ± 0.12 eV was found. The precise determination of the band alignment of Al2O3/Zn0.8Al0.2O heterojunction is of particular importance for gaining insight to the design of various electronic devices based on such heterointerface.

Excitation and propagation of X-ray fluorescence through thin devices with hollowed ordered structures: comparison of experimental and theoretical spectra.

The lack of models describing the propagation of X-rays in waveguides and the interference mechanism between incident and reflected radiation waves hamper the understanding and the control of wave propagation phenomena occurring in many real systems. Here, experimental spectra collected at the exit of microchannel plates (MCPs) under the total X-ray reflection condition are presented. The results are discussed in the framework of a theoretical model in which the wave propagation is enhanced by the presence of a transition layer at the surface. The angular distributions of the propagating radiation at the exit of these MCPs with microchannels of ∼3 µm diameter will also be presented and discussed. These spectra show contributions associated with the reflection of the primary monochromatic beam and with the fluorescence radiation originating from the excitation of atoms composing the surface of the microchannel. The soft X-ray fluorescence spectra collected at the exit of microcapillaries were analyzed in the framework of a wave approximation while diffraction contributions observed at the exit of these hollow X-ray waveguides have been calculated using the Fraunhofer diffraction model for waves in the far-field domain. Data collected at the Si L-edge show that in glassy MCPs the fluorescence radiation can be detected only when the energy of the primary monochromatic radiation is above the absorption edge for grazing angles higher than half of the critical angle of the total reflection phenomenon. Experimental data and simulations of the propagating radiation represent a clear experimental confirmation of the channeling phenomenon of the excited fluorescence radiation inside a medium and point out that a high transmission can be obtained in waveguide optics for parameters relevant to X-ray imaging.

Nano-oxide thin films deposited via atomic layer deposition on microchannel plates.

Microchannel plate (MCP) as a key part is a kind of electron multiplied device applied in many scientific fields. Oxide thin films such as zinc oxide doped with aluminum oxide (ZnO:Al2O3) as conductive layer and pure aluminum oxide (Al2O3) as secondary electron emission (SEE) layer were prepared in the pores of MCP via atomic layer deposition (ALD) which is a method that can precisely control thin film thickness on a substrate with a high aspect ratio structure. In this paper, nano-oxide thin films ZnO:Al2O3 and Al2O3 were prepared onto varied kinds of substrates by ALD technique, and the morphology, element distribution, structure, and surface chemical states of samples were systematically investigated by scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDS), X-ray diffraction (XRD), and X-ray photoemission spectroscopy (XPS), respectively. Finally, electrical properties of an MCP device as a function of nano-oxide thin film thickness were firstly studied, and the electrical measurement results showed that the average gain of MCP was greater than 2,000 at DC 800 V with nano-oxide thin film thickness approximately 122 nm. During electrical measurement, current jitter was observed, and possible reasons were preliminarily proposed to explain the observed experimental phenomenon.

Enhanced detection of high-mass proteins by using an active pixel detector.

Flying high: Application of an active pixel detector with high charge sensitivity to a linear time-of-flight mass spectrometer results in enhanced detection of high-mass proteins (such as Immunoglobulin G; IgG) using a conventional microchannel plate detection system. This technique thus provides a means to extend the mass range of such detectors as well as allowing direct visualization of mass-dependent ion-focusing phenomena.

Progress towards a 256 channel multi-anode microchannel plate photomultiplier system with picosecond timing.

Despite the rapid advances in solid state technologies such as the silicon photomultiplier (SiPM), microchannel plate (MCP) photomultipliers still offer a proven and practical technological solution for high channel count pixellated photon-counting systems with very high time resolution. We describe progress towards a 256 channel optical photon-counting system using CERN-developed NINO and HTDC ASICs, and designed primarily for time resolved spectroscopy in life science applications. Having previously built and demonstrated a 18 mm diameter prototype tube with an 8×8 channel readout configuration and <43 ps rms single photon timing resolution, we are currently developing a 40 mm device with a 32×32 channel readout. Initially this will be populated with a 256 channel electronics system comprising four sets of modular 64 channel preamplifier/discriminator, and time-to-digital converter units, arranged in a compact three dimensional configuration. We describe the detector and electronics design and operation, and present performance measurements from the 256 channel development system. We discuss enhancements to the system including higher channel count and the use of application specific on-board signal processing capabilities.

Fluorescence lifetime microscopy with a time- and space-resolved single-photon counting detector.

We have recently developed a wide-field photon-counting detector (the H33D detector) having high-temporal and high-spatial resolutions and capable of recording up to 500,000 photons per sec. Its temporal performance has been previously characterized using solutions of fluorescent materials with different lifetimes, and its spatial resolution using sub-diffraction objects (beads and quantum dots). Here we show its application to fluorescence lifetime imaging of live cells and compare its performance to a scanning confocal TCSPC approach. With the expected improvements in photocathode sensitivity and increase in detector throughput, this technology appears as a promising alternative to the current lifetime imaging solutions.

A space- and time-resolved single photon counting detector for fluorescence microscopy and spectroscopy.

We have recently developed a wide-field photon-counting detector having high-temporal and high-spatial resolutions and capable of high-throughput (the H33D detector). Its design is based on a 25 mm diameter multi-alkali photocathode producing one photo electron per detected photon, which are then multiplied up to 107 times by a 3-microchannel plate stack. The resulting electron cloud is proximity focused on a cross delay line anode, which allows determining the incident photon position with high accuracy. The imaging and fluorescence lifetime measurement performances of the H33D detector installed on a standard epifluorescence microscope will be presented. We compare them to those of standard single-molecule detectors such as single-photon avalanche photodiode (SPAD) or electron-multiplying camera using model samples (fluorescent beads, quantum dots and live cells). Finally, we discuss the design and applications of future generation of H33D detectors for single-molecule imaging and high-throughput study of biomolecular interactions.