Information-Rich Photon Imaging of Cells
Lead Research Organisation:
University of Manchester
Department Name: School of Health Sciences
Abstract
Cells and tissue are complex materials whose heterogeneity and dynamic changes are principally driven at the micron and sub-micron size scales, with underlying events ranging upwards from microsecond time scales, and measurement also based upon faster photo-electronic events of molecules (eg fluorescent and Raman emission from picosecond to nanosecond timescales). Advances in high energy physics and space research have made possible a fast photon imaging detector with >1000 channels each measuring individual photon arrivals to picosecond temporal sensitivity. We aim to develop and use this imaging detector to capture the high spatial and temporal complexity of cells and tissue. In particular, our objectives concern maximising the photon information available to unravel complex multi-component signals, including within background noise, below the visibility of present measurement and automated platforms, which underlie challenges in quantitative imaging and bioassays of live cells and tissue.
Technical Summary
The Space Research Centre at the University of Leicester have developed an Image Charge approach to read photon detections from image intensifiers onto electrodes. A multi-layer ceramic substrate with silicon resistive layer will be developed with a small pore size multi-channel plate intensifier to provide high temporal sensitivity and spatial resolution. CERN have developed an ultra fast front-end preamplifier-discriminator chip (NINO) for a precision time Time-Of-Flight detector for the Large Hadron Collider (LHC). The performance of Image Charge can be matched by measurement of the induced charge footprints of photons by the NINO chip. A high channel density NINO readout will be scaled to 1024 (32 x 32) photon measurement channels each with <25 ps photon timing sensitivity. The University of Manchester have investigated non-stop processing of the photon train detected following laser pulses (delta) and continuous wave (CW) illumination. The non-linear distribution of the photon delay times indicates that only small bunches (typically 2-4 photons) are detected with delay times shorter than the recovery time of detectors. We expect that the proposed 1024 channel photon timing device would enable time-resolved spectroscopy from both delta- and CW illumination at multiple spatial locations. This would allow presently separate time-resolved (eg fluorescent lifetime, single molecule fluorescence, Raman) and new measurements to be performed using the same detector device. Given increased information, the ability to de-convolute multiple signals will be investigated using multi-exponential fitting algorithms developed for encryption, which have enabled free and bound fluor signals to be resolved. The imaging detector will be evaluated for multiplexed bioassays and multi-parametric imaging, collaborating with Gray Cancer Institute to benefit from advanced platforms for single cell cytometry and tissue bioimaging.
People |
ORCID iD |
David Clarke (Principal Investigator) |
Publications
Hamrang Z
(2014)
Quantitative assessment of p-glycoprotein expression and function using confocal image analysis.
in Microscopy and microanalysis : the official journal of Microscopy Society of America, Microbeam Analysis Society, Microscopical Society of Canada
Hamrang Z
(2014)
Monitoring the kinetics of CellTraceā¢ calcein red-orange AM intracellular accumulation with spatial intensity distribution analysis.
in Biochimica et biophysica acta
Hamrang Z
(2012)
Raster image correlation spectroscopy as a novel tool for the quantitative assessment of protein diffusional behaviour in solution.
in Journal of pharmaceutical sciences
Hamrang Z
(2014)
Real-time evaluation of aggregation using confocal imaging and image analysis tools.
in The Analyst
Offerman S
(2014)
Ability of co-administered peptide liposome nanoparticles to exploit tumour acidity for drug delivery
in RSC Adv.
Zahra Hamrang
(2012)
Quantification and analysis of P-glycoprotein expression using spatial intensity distribution analysis
in Proceedings European Microscopy Congress
Description | Optical imaging eg microscopy allows different colours and their intensities to be seen in each pixel. The arrival times of the individual photons contributing to the light intensities in each pixel are now also seen to analyse the movement and photon emission cycle time of particles or molecules within the pixel. |
Exploitation Route | The photon correlation image analysis software is available for use in confocal microscopy and multi-photon biomaging. A photon counting detector array developed by collaborators to detect arrival times of single photons is now available from Photek as a beta test product. The ability to detect the arrival times of multiple photons at each pixel would improve on the speed and information available from such detectors. |
Sectors | Aerospace, Defence and Marine,Electronics,Healthcare,Pharmaceuticals and Medical Biotechnology,Security and Diplomacy,Transport |
Description | The photon correlation image analysis software developed has been made available to researchers. An array detector and associated electronics for photon correlation suitable for capturing such image data has been developed into a beta testing product by Photek, collaborating in the wider project with CERN and the University of Leicester. |
First Year Of Impact | 2014 |
Sector | Electronics,Pharmaceuticals and Medical Biotechnology |
Title | Analysis of a sample to determine its characteristic cycle time |
Description | An analysis for determining a characteristic cycle time of a sample. Active elements in the sample are excited with sufficient intensity and duration larger than the characteristic cycle time that at least some of the active elements are re-excited to an excited state substantially immediately following relaxation to a ground state, detecting quanta emitted by the active elements in the sample to obtain a detected signal, and analyzing the detected signal to derive the characteristic cycle time. The number of active elements in the sample and the intensity of the excitation are such that quanta are detected in a stream in which individual quanta are distinguishable from each other. |
IP Reference | US7098039 |
Protection | Patent granted |
Year Protection Granted | 2006 |
Licensed | Yes |
Impact | A detector product has been developed by Photek and made available for best testing in 2014, which enables further applications development. |