fluorescence-lifetime-macro-imager-for-biomedical-applications

Fluorescence Lifetime Macro Imager for Biomedical Applications

This paper presents a new photoluminescence lifetime imager designed to map the molecular oxygen (O2) concentration in different phosphorescent samples ...

Brookhaven National Laboratory

TimepixCam is a novel fast optical imager based on an optimized silicon pixel sensor with a thin entrance window and read out by a Timepix Application Specific Integrated Circuit. The 256 × 256 pixel sensor has a time resolution of 15 ns at a sustained frame rate of 10 Hz. We used this sensor in combination with an image intensifier for wide-field time-correlated single photon counting imaging. We have characterised the photon detection capabilities of this detector system and employed it on a wide-field epifluorescence microscope to map phosphorescence decays of various iridium complexes with lifetimes of about 1 μs in 200 μm diameter polystyrene beads.

Photon counting phosphorescence lifetime imaging with TimepixCam.

We demonstrate a coincidence velocity map imaging apparatus equipped with a novel time-stamping fast optical camera, Tpx3Cam, whose high sensitivity and nanosecond timing resolution allow for simultaneous position and time-of-flight detection. This single detector design is simple, flexible, and capable of highly differential measurements. We show detailed characterization of the camera and its application in strong field ionization experiments.

Coincidence velocity map imaging using Tpx3Cam, a time stamping optical camera with 1.5 ns timing resolution.

The application of a novel fast optical-imaging camera, TimepixCam, to molecular photoionization experiments using the velocity-map imaging technique at a free-electron laser is described. TimepixCam is a 256 × 256 pixel CMOS camera that is able to detect and time-stamp ion hits with 20 ns timing resolution, thus making it possible to record ion momentum images for all fragment ions simultaneously and avoiding the need to gate the detector on a single fragment. This allows the recording of significantly more data within a given amount of beam time and is particularly useful for pump-probe experiments, where drifts, for example, in the timing and pulse energy of the free-electron laser, severely limit the comparability of pump-probe scans for different fragments taken consecutively. In principle, this also allows ion-ion covariance or coincidence techniques to be applied to determine angular correlations between fragments.

Time-resolved ion imaging at free-electron lasers using TimepixCam.

The properties of a novel ultra-fast optical imager, Tpx3Cam, were investigated for macroscopic wide-field phosphorescent lifetime imaging (PLIM) applications. The camera is based on a novel optical sensor and Timepix3 readout chip with a time resolution of 1.6 ns, recording of photon arrival time and time over threshold for each pixel, and readout rate of 80 megapixels per second. In this study, we coupled the camera to an image intensifier, a 760 nm emission filter and a 50 mm lens, and with a super-bright 627nm LED providing pulsed excitation of a 18 × 18 mm sample area. The resulting macro-imager with compact and rigid optical alignment of its main components was characterised using planar phosphorescent O sensors and a resolution plate mask. Several acquisition and image processing algorithms were evaluated to optimise the system resolution and performance for the wide-field PLIM, followed by imaging a variety of phosphorescent samples. The new PLIM system looks promising, particularly for phosphorescence lifetime-based imaging of O in various chemical and biological samples.

New luminescence lifetime macro-imager based on a Tpx3Cam optical camera.

A camera-based three-dimensional (3D) imaging system with a superb time-of-flight (TOF) resolution and multi-hit capability was recently developed for electron/ion imaging [Lee et al. J. Chem. Phys. 141, 221101 (2014)]. In this work, we report further improvement of the event rate of the system by adopting an event-driven camera, Tpx3Cam, for detecting the 2D positions of electrons, while a high-speed digitizer provides highly accurate (∼30 ps) TOF information for each event at a rate approaching 1 Mhits/sec.

Developing a camera-based 3D momentum imaging system capable of 1 Mhits/s.

Scalable technologies to characterize the performance of quantum devices are crucial to creating large quantum networks and quantum processing units. Chief among the resources of quantum information processing is entanglement. Here we describe the full temporal and spatial characterization of polarization-entangled photons produced by Spontaneous Parametric Down Conversions using an intensified high-speed optical camera, Tpx3Cam. This novel technique allows for precise determination of Bell inequality parameters with minimal technical overhead, and for new characterization methods for the spatial distribution of entangled quantum information. The fast-optical camera could lead to multiple applications in Quantum Information Science, opening new perspectives for the scalability of quantum experiments.

Fast camera spatial characterization of photonic polarization entanglement.

The uses of a silicon-pixel camera with very good time resolution (∼nanosecond) for detecting multiple, bunched optical photons is explored. We present characteristics of the camera and describe experiments proving its counting capabilities. We use a spontaneous parametric down-conversion source to generate correlated photon pairs, and exploit the Hong-Ou-Mandel (HOM) interference effect in a fiber-coupled beam splitter to bunch the pair onto the same output fiber. It is shown that the time and spatial resolution of the camera enables independent detection of two photons emerging simultaneously from a single spatial mode.

Counting of Hong-Ou-Mandel Bunched Optical Photons Using a Fast Pixel Camera.

O PLIM microscopy was employed in various studies, however current platforms have limitations in sensitivity, image acquisition speed, accuracy and general usability. We describe a new PLIM imager based on the Timepix3 camera (Tpx3cam) and its application for imaging of O concentration in various tissue samples stained with a nanoparticle based probe, NanO2-IR. Upon passive staining of mouse brain, lung or intestinal tissue surface with minute quantities of NanO2-IR or by microinjecting the probe into the lumen of small or large intestine fragments, robust phosphorescence intensity and lifetime signals were produced, which allow mapping of O in the tissue within 20 s. Inhibition of tissue respiration or limitation of O diffusion to tissue produced the anticipated increases or decreases in O levels, respectively. The difference in O concentration between the colonic lumen and air-exposed serosal surface was around 140 µM. Furthermore, subcutaneous injection of 5 µg of the probe in intact organs (a paw or tail of sacrificed mice) enabled efficient O imaging at tissue depths of up to 0.5 mm. Overall, the PLIM imager holds promise for metabolic imaging studies with various ex vivo models of animal tissue, and also for use in live animals.

Mapping O concentration in ex-vivo tissue samples on a fast PLIM macro-imager.

We demonstrate three-dimensional velocity map imaging of low energy electrons using a TPX3CAM, where the three-dimensional momentum information [p, p, p] is encoded in position and timing [x, y, t] of hits on the camera sensor. We make use of the camera sensor for the [x, y] information and a constant fraction discriminator and fast time to digital converter in the camera for the time information. We illustrate the capabilities of our apparatus by presenting above threshold ionization measurements of xenon, which produces well defined structures in the momentum resolved photoelectron yield.

3D velocity map imaging of electrons with TPX3CAM.

We present a simple approach to characterize the spatial variation of the gain in microchannel plate (MCP) coupled to phosphor detectors using single electron or photon hits. The technique is easy to implement and general enough to be extended to other kinds of detectors. We demonstrate the efficacy of the approach on both laboratory and Monte Carlo generated datasets. Furthermore, we use the approach to measure the variation in gain over time as the MCP is exposed to an increasing number of electrons.

A simple approach for characterizing the spatially varying sensitivity of microchannel plate detectors.

Ion microscopy allows for high-throughput mass spectrometry imaging. In order to resolve congested mass spectra, a high degree of timing precision is required from the microscope detector. In this paper we present an ion microscope mass spectrometer that uses a Timepix3 hybrid pixel readout with an optimal 1.56 ns resolution. A novel triggering technique is also employed to remove the need for an external time-to-digital converter (TDC) and allow the experiment to be performed using a low-cost and commercially available readout system. Results obtained from samples of rhodamine B demonstrate the application of multimass imaging sensors for microscope mass spectrometry imaging with high mass resolution.

Andrei Nomerotski

Physics Department

Brookhaven National Laboratory

Fluorescence Lifetime Macro Imager for Biomedical Applications

Andrei Nomerotski

.css-o250i3{font-size:18px;font-family:Open Sans,sans-serif;line-height:21.6px;}Physics Department

Brookhaven National Laboratory

Fluorescence Lifetime Macro Imager for Biomedical Applications

Physics Department