Information-Rich Photon Imaging of Cells

Lead Research Organisation: University of Leicester
Department Name: Physics and Astronomy

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.

Publications

10 25 50
publication icon
Brook N (2018) Testbeam studies of a TORCH prototype detector in Nuclear Instruments and Methods in Physics Research Section A: Accelerators, Spectrometers, Detectors and Associated Equipment

publication icon
Despeisse M (2011) Multi-Channel Amplifier-Discriminator for Highly Time-Resolved Detection in IEEE Transactions on Nuclear Science

publication icon
Lapington J (2009) A multi-channel high time resolution detector for high content imaging in Nuclear Instruments and Methods in Physics Research Section A: Accelerators, Spectrometers, Detectors and Associated Equipment

publication icon
Lapington J (2011) Multi-channel picosecond photon timing with microchannel plate detectors in Nuclear Instruments and Methods in Physics Research Section A: Accelerators, Spectrometers, Detectors and Associated Equipment

publication icon
Lapington J (2009) A high-throughput, multi-channel photon-counting detector with picosecond timing in Nuclear Instruments and Methods in Physics Research Section A: Accelerators, Spectrometers, Detectors and Associated Equipment

publication icon
Lapington JS (2012) Progress towards a 256 channel multi-anode microchannel plate photomultiplier system with picosecond timing. in Nuclear instruments & methods in physics research. Section A, Accelerators, spectrometers, detectors and associated equipment

publication icon
Milnes J (2014) The TORCH PMT, a close packing, multi-anode, long life MCP-PMT for Cherenkov applications in Nuclear Instruments and Methods in Physics Research Section A: Accelerators, Spectrometers, Detectors and Associated Equipment

 
Description The goal of this project was to develop a fast photon imaging detector with >1000 channels measuring individual photon arrivals to picosecond accuracy. The project built on an existing, STFC-funded small format proof-of-concept demonstrator which proved the feasibility of utilizing very high speed multi-channel ASIC electronics, originally developed for the LHC at CERN, in a multi-anode microchannel plate photomultiplier.

This project developed the demonstrator detector into a fully featured prototype, enhancing its performance, channel count, and practicality of use in a life science laboratory. The system currently has no effective competition offering this level of multi-channel capability with comparable time resolution and event throughput in a compact, integrated device. The major developments made within this project were:-

1. Development of a state of the art 1024 channel multi-anode microchannel plate detector with integrated 32 x 32 pixel2 readout format implemented on a multi-layer ceramic component providing vacuum integrity and 1024 channel vacuum electrical feed-through. Prototype measured performance of photon event timing of 43 ps per channel (precision limited by laser test equipment) and photon maximum count rate of 10 Mcount/s/cm2.

2. Development of modular, miniature integrated electronics suite comprising i) high density 1024 channel multi-anode interconnection, ii) 64 channel preamplifier/discriminator and time-to-digital converter modules, iii) backplane to accommodate up to 16 modules and programmable digital processing board with USB interface. Electronics count rate performance of 2.5Mcount/s/ch.

3. Integration of detector and electronics into a small footprint package with dimensions of 150x100x100 mm3 with suitable interfaces for retro-fitting to biological microscope instrumentation and a widely available USB interface for interface to supporting computer software.

We have manufactured two complete 40 mm 256 channel camera systems with 100 ps time resolution and standard C-mount interfaces (channel number and time resolution of demonstrator systems were descoped owing to schedule and cost limitations). These will be available as demonstrators to allow evaluation of the system on potential end-user equipment.

Our industrial collaborators, Photek, have now (2018) developed a square format detector using the technologies developed in the IRPICS project, for the TORCH Cherenkov detector upgrade for LHCb. They have also released a version of the pixellated readout IRPICS detector onto the market as a commercial product.

The IRPICS readout anode combined with an Active Anode technique is now being used (2019) with a timing ASIC, TOFPET2, for a picosecond timing high spatial resolution microchannel plate detector in collaboration with Photek Ltd. THe new detector concept, utilising a pixellated image readout with charge centroiding has applications in space science (UV astronomy, auroral imaging, space weather) and fast timing commercial opportunities. A poster was presented at the Vienna Conference on Instrumentation 2019 and a publication is in preparation.
Exploitation Route Non-academic uses of the High speed detector system
1. Clinical diagnostics and healthcare
a. High speed flow cytometry techniques
b. Imaging techniques
i. Confocal microscopy
ii. TOFPET
iii. Optical tomography
c. High Content techniques using time resolved spectroscopies, e.g. drug screening
2. Environmental and remote imaging
a. LIDAR from space
3. Materials science
a. Surface analysis
b. Mass spectrometry

Non-academic uses of the Modular electronics system

1. Application to alternative detector technologies such as APD arrays, SiPM devices, delay line readouts, for a variety of commercial applications. Potential applications in both research and commercial sectors:
1. High throughput time-correlated single photon counting techniques e.g. Fluorescence lifetime imaging (FLIM).
2. Fluorescence correlation spectroscopy.
3. Flow cytometry:
4. Optical tomography.
5. Confocal microscopy.
6. LIDAR.
7. Time-of-flight techniques including
a. TOFPET
b. Field ion microscopy - e.g. the Atom Probe.
Application opportunities being followed up:
1. FLIM: field trials on a FLIM research microscope developed by Professor Boris Vojnovic at the Gray Institute for radiation Oncology, University of Oxford.
2. Confocal microscopy: We have been awarded innovation fellowship funding from the University of Leicester to develop a proof-of-concept demonstration of the detector system for another BBSRC TDRI-funded technology: a novel confocal microscopy technique application utilizing a digital micro-mirror device, with the expectation of follow-on funding for further development.

We are currently negotiating a new licensing agreement with Photek Ltd. for commercialization of the products already developed within the project, and new ones developed as a result of the project collaboration.
Products developed within the project have immediate commercial potential:
1. A 40 mm 256 channel camera system with 100 ps time resolution and standard C-mount interfaces with research and commercial potential and applications in life science and other fields.
2. The system electronics developed within the project is based on a modular design and can be configured for channels densities count from 64 up to the current accommodation limit of 1024 channels. This is being made commercially available from Photek for alternative detector technologies such as APD arrays, SiPM devices, delay line readouts, etc.
3. A complementary KTP project in collaboration with Photek and funded jointly by STFC and TSB, has utilized the electronics developed within the for a high resolution imaging detector with sub-100 ps event timing with commercial applications in the life sciences and other fields. The KTP Associate is also about to complete a PhD course at the University of Leicester
Other academic and commercial potential being pursued:
1. Time resolved Raman spectroscopy (TRRS): We are in collaboration with IS-Instruments Ltd., a company specializing in Raman spectroscopy to utilise the IRPICS electronics system with a silicon photomultiplier array for TRRS. We have already been awarded TSB funding for a feasibility study and are also collaborating with the University of Sheffield and IS-Instruments in an STFC IPS proposal to develop a custom detector using the IRPICS electronics for a commercial TRRS system
2. Field ion microscopy - e.g. the Atom Probe. We are consulting with Professor George Smith and Dr Michael Moody, Materials Department, Oxford University, on application of the detector system
3. The development of high speed Cherenkov detectors for Nuclear and Particle physics experiments, e.g. the PANDA experiment on FAIR, and RICH detectors on the TORCH LHCb experiments at CERN. Our industrial collaborator Photek is now a leading player in supply of high time resolution MCP-based detectors for these fields.
Sectors Aerospace

Defence and Marine

Electronics

Environment

Healthcare

Pharmaceuticals and Medical Biotechnology

Security and Diplomacy

Other

 
Description Within the Information Rich Photon Imaging of Cells grant, we developed a photodetector for high content analyses including fluorescence lifetime imaging at very high throughput. The detector design broke new ground in several areas: 1. it used a multi-layer ceramic anode to provide a compact, vacuum compatible image readout device with high density readout. 2. It funded development of a modified preamplifier - discriminator chip (NINO32) with high channel density to allow a compact modular electronics system to be built 3. It resulted in the design and manufacture of a modular electronics system of up to 1024 independent channels, with very high throughput (up to 2MHz per channel) and time resolution down to 25 picosecond. These technologies have been taken up in several new areas: 1. The multi-layer ceramic readout technology has given rise to the production by Photek Ltd. of a new, square format imaging photomultiplier, initially developed for the TORCH FP7 project, to develop a new Ring Imaging Cherenkov detector for the LHCb upgrade at CERN. This technology will also have application in similar instruments within particle and nuclear physics e.g. at FAIR, etc., and in the broader commercial field with application to areas including time correlated single photon counting for biological microscopy, drug discovery, and high content bioassay. A successful product for these markets will increase the economic competitiveness of both Photek and related UK industries, and applications of the instrument in the biomedical and health sectors have the capability to enhance health and well-being of the populace., 2. The modular multi-channel high time resolution electronics is now being applied to silicon photomultiplier devices as a cost-effective route to a portable, commercial system for time resolved Raman spectroscopy in conjunction with IS-Instruments Ltd.. Such as system will provide new opportunities in fields such as security for detection of illegal drugs and explosive, detection of counterfeit drugs, and medical applications. The economic and societal impacts of this technology are potentially widespread. A cost-effective, time-resolved Raman spectrometer would open up an untapped worldwide market for Raman spectroscopy applications where performance (particularly fluorescence rejection), affordability, and portability are necessary requirements; such a market would be open for exploitation by UK industry. The European suppliers of key component parts such as high rep rate lasers and diffraction gratings will also benefit from increased sales. All these factors will have positive impact to the local, UK and European economies. Recent collaboration (2016 onwards) with Petsys S.A. and Photek Ltd. have led to further development of the IRPICS detector sustem with an all new, more cost effective and compact electronics. We are actively seeking to commercialise this reviosed IRPICS system and its successor, which will have a higher resolution readout. Photek have also now developed a square format photomultiplier as a commercial product, based around the original IRPICS design, with low deadspace which allows mosaicing of IRPICS detectors for large readout format requirements. One such is the TORCH CHerenkov detector being developed for the CERN LHCb upgrade. Pixellated microchannel plate detectors developed from the original IRPICS concept have now been demonstrated with fast timing ASICs to enable picosecond photon imaging in collaboration with Photek Ltd. 2018: A product has been launched covering a broad range of research and commercial applications, from particle physics to healthcare.
First Year Of Impact 2011
Sector Aerospace, Defence and Marine,Electronics,Environment,Healthcare,Pharmaceuticals and Medical Biotechnology,Security and Diplomacy,Other
Impact Types Societal

Economic

 
Description Seeing more than before: emerging imaging technologies Competition February 2014
Amount £49,992 (GBP)
Funding ID 41462-312150 
Organisation Innovate UK 
Sector Public
Country United Kingdom
Start 11/2014 
End 10/2015
 
Description Technology Strategy Board Knowledge Transfer Programme
Amount £202,000 (GBP)
Organisation Innovate UK 
Sector Public
Country United Kingdom
Start 03/2009 
End 03/2012
 
Description University of Leicester Proof of Concept Fund - High speed confocal microscopy
Amount £10,000 (GBP)
Organisation University of Leicester 
Sector Academic/University
Country United Kingdom
Start 01/2011 
End 12/2011
 
Description University of Leicester Proof of Concept Fund - Time resolved Raman Spectroscopy
Amount £14,800 (GBP)
Organisation University of Leicester 
Sector Academic/University
Country United Kingdom
Start 03/2014 
End 08/2014
 
Description IRPICS project collaboration 
Organisation European Organization for Nuclear Research (CERN)
Country Switzerland 
Sector Academic/University 
PI Contribution Academic and commercial partners of the IRPICS project collaboration
Start Year 2006
 
Description IRPICS project collaboration 
Organisation Photek Ltd.
Country United Kingdom 
Sector Private 
PI Contribution Academic and commercial partners of the IRPICS project collaboration
Start Year 2004
 
Description IRPICS project collaboration 
Organisation University of Manchester
Country United Kingdom 
Sector Academic/University 
PI Contribution Academic and commercial partners of the IRPICS project collaboration
Start Year 2006
 
Description IRPICS project collaboration 
Organisation University of Oxford
Country United Kingdom 
Sector Academic/University 
PI Contribution Academic and commercial partners of the IRPICS project collaboration
Start Year 2006
 
Description TOFPET 
Organisation PETsys Electronics S.A.
Country Portugal 
Sector Private 
PI Contribution Evaluation of a multi-channel fast timing electronics system for applications requiring time resolved photon-counting spectroscopy.
Collaborator Contribution Collaboration in development of electronics and detectors for a photon-counting SPAD array for time resolved photon-counting spectroscopy.
Impact Development of a demonstrator system for prior to commercialisation. Report into performance of a multi-channel fast timing electronics system developed by Petsys electronics - medical pet detectors, s.a.
Start Year 2016
 
Description TOFPET 
Organisation Photek Ltd.
Country United Kingdom 
Sector Private 
PI Contribution Evaluation of a multi-channel fast timing electronics system for applications requiring time resolved photon-counting spectroscopy.
Collaborator Contribution Collaboration in development of electronics and detectors for a photon-counting SPAD array for time resolved photon-counting spectroscopy.
Impact Development of a demonstrator system for prior to commercialisation. Report into performance of a multi-channel fast timing electronics system developed by Petsys electronics - medical pet detectors, s.a.
Start Year 2016
 
Description Time-resolved Spectroscopy 
Organisation Delft University of Technology (TU Delft)
Country Netherlands 
Sector Academic/University 
PI Contribution Expertise in design and operation of high speed photon-counting linear detector arrays and electronics for time-resolved Raman spectroscopy. Assembly, integration, verification and calibration of high speed photon-counting detector systems.
Collaborator Contribution IS-INstruments: Expertise in spectrometer design and operation, market study, product commercialization and marketing University of Sheffield: Expertise in IR and optical SPAD array design and manufacture. University College Cork: Expertise in silicon single photon avalanche detector design and manufacture University of Delft: Design and manufacture of SPAD arrays and timing electronics EPFL: Design and manufacture of SPAD arrays and timing electronics
Impact Funding achieved: InnovateUK "Emerging Imaging technologies" proof-of-concept award - £150k STFC IPS award - £450k
Start Year 2014
 
Description Time-resolved Spectroscopy 
Organisation IS Instruments
Country United Kingdom 
Sector Private 
PI Contribution Expertise in design and operation of high speed photon-counting linear detector arrays and electronics for time-resolved Raman spectroscopy. Assembly, integration, verification and calibration of high speed photon-counting detector systems.
Collaborator Contribution IS-INstruments: Expertise in spectrometer design and operation, market study, product commercialization and marketing University of Sheffield: Expertise in IR and optical SPAD array design and manufacture. University College Cork: Expertise in silicon single photon avalanche detector design and manufacture University of Delft: Design and manufacture of SPAD arrays and timing electronics EPFL: Design and manufacture of SPAD arrays and timing electronics
Impact Funding achieved: InnovateUK "Emerging Imaging technologies" proof-of-concept award - £150k STFC IPS award - £450k
Start Year 2014
 
Description Time-resolved Spectroscopy 
Organisation Swiss Federal Institute of Technology in Lausanne (EPFL)
Country Switzerland 
Sector Public 
PI Contribution Expertise in design and operation of high speed photon-counting linear detector arrays and electronics for time-resolved Raman spectroscopy. Assembly, integration, verification and calibration of high speed photon-counting detector systems.
Collaborator Contribution IS-INstruments: Expertise in spectrometer design and operation, market study, product commercialization and marketing University of Sheffield: Expertise in IR and optical SPAD array design and manufacture. University College Cork: Expertise in silicon single photon avalanche detector design and manufacture University of Delft: Design and manufacture of SPAD arrays and timing electronics EPFL: Design and manufacture of SPAD arrays and timing electronics
Impact Funding achieved: InnovateUK "Emerging Imaging technologies" proof-of-concept award - £150k STFC IPS award - £450k
Start Year 2014
 
Description Time-resolved Spectroscopy 
Organisation University College Cork
Department School of Electrical and Electronic Engineering
Country Ireland 
Sector Academic/University 
PI Contribution Expertise in design and operation of high speed photon-counting linear detector arrays and electronics for time-resolved Raman spectroscopy. Assembly, integration, verification and calibration of high speed photon-counting detector systems.
Collaborator Contribution IS-INstruments: Expertise in spectrometer design and operation, market study, product commercialization and marketing University of Sheffield: Expertise in IR and optical SPAD array design and manufacture. University College Cork: Expertise in silicon single photon avalanche detector design and manufacture University of Delft: Design and manufacture of SPAD arrays and timing electronics EPFL: Design and manufacture of SPAD arrays and timing electronics
Impact Funding achieved: InnovateUK "Emerging Imaging technologies" proof-of-concept award - £150k STFC IPS award - £450k
Start Year 2014
 
Description Time-resolved Spectroscopy 
Organisation University of Sheffield
Country United Kingdom 
Sector Academic/University 
PI Contribution Expertise in design and operation of high speed photon-counting linear detector arrays and electronics for time-resolved Raman spectroscopy. Assembly, integration, verification and calibration of high speed photon-counting detector systems.
Collaborator Contribution IS-INstruments: Expertise in spectrometer design and operation, market study, product commercialization and marketing University of Sheffield: Expertise in IR and optical SPAD array design and manufacture. University College Cork: Expertise in silicon single photon avalanche detector design and manufacture University of Delft: Design and manufacture of SPAD arrays and timing electronics EPFL: Design and manufacture of SPAD arrays and timing electronics
Impact Funding achieved: InnovateUK "Emerging Imaging technologies" proof-of-concept award - £150k STFC IPS award - £450k
Start Year 2014
 
Title Multi-channel modular photon counting electronics with 25 picosecond time resolution 
Description A modular electronics system built around NINO and HPTDC ASICs developed at CERN for LHC-ALICE. Modules can be assembled into a system comprising up to 1024 channels, each capable of up to 2MHz throughput and 25 picosecond time resolution. 
Type Of Technology Detection Devices 
Year Produced 2010 
Impact These electronics were developed for fluorescence lifetime imaging and other time resolved spectroscopy applications original in conjunction with a microchannel plate photomultiplier. They are now being used to pioneer a new technique for time resolved Raman spectroscopy using a silicon photomultiplier array and Fourier transform spectrometer. This portable system will have application in security for drug and explosive detection, detection of counterfeit drugs, and medical applications. 2015: System sold to Swiss security systems commercial developer for evalution 2016: In discussion with US contractor for use in planetary LIDAR instrumentation 
 
Title Multi-layer ceramic pixellated readout devices for imaging photomultipliers 
Description A multi-layer ceramic device combining an image readout array and multi-channel electrical feed-through for imaging photo-multipliers. The device is UHV compatible and provides vacuum containment as well as high density multi-channel electrical feedthroughs (~1024 channels) 
Type Of Technology Detection Devices 
Year Produced 2010 
Impact This device permits manufacture of a compact but very high density image readout for photo-multipliers capable of high speed readout and picosecond timing resolution. It forms the basis of a new square format, high fill factor, imaging photomultiplier being manufactured for the TORCH project, the development of a new Ring Imaging Cherenkov detector for the LHCb upgrade at CERN. 
 
Description Medical imaging - Opportunities for Business Workshop 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Type Of Presentation Paper Presentation
Geographic Reach International
Primary Audience Participants in your research or patient groups
Results and Impact Presentation of IRPICS technologies to a workshop organised by the Space Ideas Hub based at the University of Leicester for East Midlands industries in the life science field.

no actual impacts realised to date
Year(s) Of Engagement Activity 2012