Imperial College London

DrPeterQuicke

Faculty of EngineeringDepartment of Bioengineering

Visiting Researcher
 
 
 
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peter.quicke10 CV

 
 
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Blackett LaboratorySouth Kensington Campus

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Summary

 

Publications

Publication Type
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22 results found

Quicke P, Sun Y, Beykou M, Arias-Garcia M, Bakal C, Djamgoz M, Acker C, Foust Aet al., 2022, Voltage imaging reveals the dynamic electrical signatures of human breast cancer cells, Communications Biology, Vol: 5, ISSN: 2399-3642

Cancer cells feature a resting membrane potential (Vm) that is depolarized compared to normal cells, and express active ionic conductances, which factor directly in their pathophysiological behavior. Despite similarities to ‘excitable’ tissues, relatively little is known about cancer cell Vm dynamics. Here high-throughput, cellular-resolution Vm imaging reveals that Vm fluctuates dynamically in several breast cancer cell lines compared to non-cancerous MCF-10A cells. We characterize Vm fluctuations of hundreds of human triple-negative breast cancer MDA-MB-231 cells. By quantifying their Dynamic Electrical Signatures (DESs) through an unsupervised machine-learning protocol, we identify four classes ranging from "noisy” to “blinking/waving“. The Vm of MDA-MB-231 cells exhibits spontaneous, transient hyperpolarizations inhibited by the voltage-gated sodium channel blocker tetrodotoxin, and by calcium-activated potassium channel inhibitors apamin and iberiotoxin. The Vm of MCF-10A cells is comparatively static, but fluctuations increase following treatment with transforming growth factor-β1, a canonical inducer of the epithelial-to-mesenchymal transition. These data suggest that the ability to generate Vm fluctuations may be a property of hybrid epithelial-mesenchymal cells or those originated from luminal progenitors.

Journal article

Howe C, Song P, Verinaz Jadan HI, Dragotti PL, Quicke P, Foust Aet al., 2022, Comparing synthetic refocusing to deconvolution for the extraction of neuronal calcium transients from light fields, Neurophotonics, Vol: 9, Pages: 1-17, ISSN: 2329-4248

Significance: Light-field microscopy (LFM) enables fast, light-efficient, volumetric imaging of neuronal activity with calcium indicators. Calcium transients differ in temporal signal-to-noise ratio (tSNR) and spatial confinement when extracted from volumes reconstructed by different algorithms.Aim: We evaluated the capabilities and limitations of two light-field reconstruction algorithms for calcium fluorescence imaging.Approach: We acquired light-field image series from neurons either bulk-labeled or filled intracellularly with the red-emitting calcium dye CaSiR-1 in acute mouse brain slices. We compared the tSNR and spatial onfinement of calcium signals extracted from volumes reconstructed with synthetic refocusing and Richardson-Lucy 3D deconvolution with and without total variation regularization.Results: Both synthetic refocusing and Richardson-Lucy deconvolution resolved calcium signals from single cells and neuronal dendrites in three dimensions. Increasing deconvolution iteration number improved spatial confinement but reduced tSNR compared to synthetic refocusing. Volumetric light-field imaging did not decrease calcium signal tSNR compared to interleaved, widefield image series acquired in matched planes.Conclusions: LFM enables high-volume rate, volumetric imaging of calcium transients in single cells (bulk-labeled), somata and dendrites (intracellular loaded). The trade-offs identified for tSNR, spatial confinement, and computational cost indicate which of synthetic refocusing or deconvolution can better realize the scientific requirements of future LFM calcium imaging applications.

Journal article

Quicke P, Sun Y, Arias-Garcia M, Acker CD, Djamgoz MBA, Bakal C, Foust AJet al., 2021, Membrane voltage fluctuations in human breast cancer cells

<jats:title>ABSTRACT</jats:title><jats:p>Cancer cells feature a resting membrane potential (<jats:italic>V</jats:italic><jats:sub><jats:italic>m</jats:italic></jats:sub>) that is depolarized compared to normal cells, and express active ionic conductances, which factor directly in their pathophysiological behavior. Despite similarities to ‘excitable’ tissues, relatively little is known about cancer cell <jats:italic>V</jats:italic><jats:sub><jats:italic>m</jats:italic></jats:sub> dynamics. With high-throughput, cellular-resolution <jats:italic>V</jats:italic><jats:sub><jats:italic>m</jats:italic></jats:sub> imaging, we characterized <jats:italic>V</jats:italic><jats:sub><jats:italic>m</jats:italic></jats:sub> fluctuations of hundreds of human triple-negative breast cancer MDA-MB-231 cells and compared to non-cancerous breast epithelial MCF-10A cells. By quantifying their Dynamic Electrical Signatures (DESs) through an unsupervised machine-learning protocol, we identified four classes ranging from “noisy” to “blinking/waving”. The <jats:italic>V</jats:italic><jats:sub><jats:italic>m</jats:italic></jats:sub> of MDA-MB-231 cells exhibited spontaneous, transient hyperpolarizations that were inhibited by the voltage-gated sodium channel blocker tetrodotoxin. The <jats:italic>V</jats:italic><jats:sub><jats:italic>m</jats:italic></jats:sub> of MCF-10A cells was comparatively static, but fluctuations increased following treatment with transforming growth factor-β1, a canonical inducer of the epithelial-to-mesenchymal transition. These data suggest that the ability to generate <jats:italic>V</jats:italic><jats:sub><jats:italic>m</jats:italic></jats:sub> fluctuations is a

Journal article

Quicke P, Howe CL, Foust A, 2021, Balancing the fluorescence imaging budget for all-optical neurophysiology experiments, All-optical methods to study neuronal function, Editors: Papagiakoumou, Publisher: Humana Press

The goal of this chapter is to establish a framework to evaluate imaging methodologies for all-optical neurophysiology experiments. This is not an exhaustive review of fluorescent indicators and imaging modalities but rather aims to distill the functional imaging principles driving the choice of both. Scientific priorities determine whether the imaging strategy is based on an “optimal fluorescent indicator” or “optimal imaging modality.” The choice of the first constrains the choice of the second due to each’s contributions to the fluorescence budget and signal-to-noise ratio of time-varying fluorescence changes.

Book chapter

Song P, Jadan HV, Howe CL, Quicke P, Foust AJ, Dragotti PLet al., 2021, MODEL-INSPIRED DEEP LEARNING FOR LIGHT-FIELD MICROSCOPY WITH APPLICATION TO NEURON LOCALIZATION, IEEE International Conference on Acoustics, Speech and Signal Processing (ICASSP), Publisher: IEEE, Pages: 8087-8091

Conference paper

Verinaz-Jadan H, Song P, Howe CL, Quicke P, Foust AJ, Dragotti PLet al., 2021, DEEP LEARNING FOR LIGHT FIELD MICROSCOPY USING PHYSICS-BASED MODELS, 18th IEEE International Symposium on Biomedical Imaging (ISBI), Publisher: IEEE, Pages: 1091-1094, ISSN: 1945-7928

Conference paper

Howe CL, Quicke P, Song P, Jadan HV, Dragotti PL, Foust AJet al., 2020, Comparing synthetic refocusing to deconvolution for the extraction of neuronal calcium transients from light-fields

<jats:title>Abstract</jats:title><jats:sec><jats:title>Significance</jats:title><jats:p>Light-field microscopy (LFM) enables fast, light-efficient, volumetric imaging of neuronal activity with calcium indicators. Calcium transients differ in temporal signal-to-noise ratio (tSNR) and spatial confinement when extracted from volumes reconstructed by different algorithms.</jats:p></jats:sec><jats:sec><jats:title>Aim</jats:title><jats:p>We evaluated the capabilities and limitations of two light-field reconstruction algorithms for calcium fluorescence imaging.</jats:p></jats:sec><jats:sec><jats:title>Approach</jats:title><jats:p>We acquired light-field image series from neurons either bulk-labeled or filled intracellularly with the red-emitting calcium dye CaSiR-1 in acute mouse brain slices. We compared the tSNR and spatial confinement of calcium signals extracted from volumes reconstructed with synthetic refocusing and Richardson-Lucy 3D deconvolution with and without total variation regularization.</jats:p></jats:sec><jats:sec><jats:title>Results</jats:title><jats:p>Both synthetic refocusing and Richardson-Lucy deconvolution resolved calcium signals from single cells and neuronal dendrites in three dimensions. Increasing deconvolution iteration number improved spatial confinement but reduced tSNR compared to synthetic refocusing. Volumetric light-field imaging did not decrease calcium signal tSNR compared to interleaved, widefield image series acquired in matched planes.</jats:p></jats:sec><jats:sec><jats:title>Conclusions</jats:title><jats:p>LFM enables high-volume rate, volumetric imaging of calcium transients in single cells (bulk-labeled), somata and dendrites (intracellular loaded). The trade-offs identified for tSNR, spatial confinement, and computational cost indicate which of syntheti

Working paper

Quicke P, Howe CL, Song P, Jadan HV, Song C, Knöpfel T, Neil M, Dragotti PL, Schultz SR, Foust AJet al., 2020, Subcellular resolution three-dimensional light-field imaging with genetically encoded voltage indicators, Neurophotonics, Vol: 7, ISSN: 2329-4248

Significance: Light-field microscopy (LFM) enables high signal-to-noise ratio (SNR) and light efficient volume imaging at fast frame rates. Voltage imaging with genetically encoded voltage indicators (GEVIs) stands to particularly benefit from LFM's volumetric imaging capability due to high required sampling rates and limited probe brightness and functional sensitivity. Aim: We demonstrate subcellular resolution GEVI light-field imaging in acute mouse brain slices resolving dendritic voltage signals in three spatial dimensions. Approach: We imaged action potential-induced fluorescence transients in mouse brain slices sparsely expressing the GEVI VSFP-Butterfly 1.2 in wide-field microscopy (WFM) and LFM modes. We compared functional signal SNR and localization between different LFM reconstruction approaches and between LFM and WFM. Results: LFM enabled three-dimensional (3-D) localization of action potential-induced fluorescence transients in neuronal somata and dendrites. Nonregularized deconvolution decreased SNR with increased iteration number compared to synthetic refocusing but increased axial and lateral signal localization. SNR was unaffected for LFM compared to WFM. Conclusions: LFM enables 3-D localization of fluorescence transients, therefore eliminating the need for structures to lie in a single focal plane. These results demonstrate LFM's potential for studying dendritic integration and action potential propagation in three spatial dimensions.

Journal article

Quicke P, Howe CL, Song P, Jadan HV, Song C, Knöpfel T, Neil M, Dragotti PL, Schultz SR, Foust AJet al., 2020, Subcellular resolution 3D light field imaging with genetically encoded voltage indicators, Neurophotonics, Vol: 7, ISSN: 2329-4248

Significance: Light-field microscopy (LFM) enables high signal-to-noise ratio (SNR) and light efficient volume imaging at fast frame rates. Voltage imaging with genetically encoded voltage indicators (GEVIs) stands to particularly benefit from LFM’s volumetric imaging capability due to high required sampling rates and limited probe brightness and functional sensitivity.Aim: We demonstrate subcellular resolution GEVI light-field imaging in acute mouse brain slices resolving dendritic voltage signals in three spatial dimensions.Approach: We imaged action potential-induced fluorescence transients in mouse brain slices sparsely expressing the GEVI VSFP-Butterfly 1.2 in wide-field microscopy (WFM) and LFM modes. We compared functional signal SNR and localization between different LFM reconstruction approaches and between LFM and WFM.Results: LFM enabled three-dimensional (3-D) localization of action potential-induced fluorescence transients in neuronal somata and dendrites. Nonregularized deconvolution decreased SNR with increased iteration number compared to synthetic refocusing but increased axial and lateral signal localization. SNR was unaffected for LFM compared to WFM.Conclusions: LFM enables 3-D localization of fluorescence transients, therefore eliminating the need for structures to lie in a single focal plane. These results demonstrate LFM’s potential for studying dendritic integration and action potential propagation in three spatial dimensions.

Journal article

Song P, Verinaz Jadan H, Howe C, Quicke P, Foust A, Dragotti PLet al., 2020, 3D localization for light-field microscopy via convolutional sparse coding on epipolar images, IEEE transactions on computational imaging, Vol: 6, Pages: 1017-1032, ISSN: 2333-9403

Light-field microscopy (LFM) is a type of all-optical imaging system that is able to capture 4D geometric information of light rays and can reconstruct a 3D model from a single snapshot. In this paper, we propose a new 3D localization approach to effectively detect 3D positions of neuronal cells from a single light-field image with high accuracy and outstanding robustness to light scattering. This is achieved by constructing a depth-aware dictionary and by combining it with convolutional sparse coding. Specifically, our approach includes 3 key parts: light-field calibration, depth-aware dictionary construction, and localization based on convolutional sparse coding (CSC). In the first part, an observed raw light-field image is calibrated and then decoded into a two-plane parameterized 4D format which leads to the epi-polar plane image (EPI). The second part involves simulating a set of light-fields using a wave-optics forward model for a ball-shaped volume that is located at different depths. Then, a depth-aware dictionary is constructed where each element is a synthetic EPI associated to a specific depth. Finally, by taking full advantage of the sparsity prior and shift-invariance property of EPI, 3D localization is achieved via convolutional sparse coding on an observed EPI with respect to the depth-aware EPI dictionary. We evaluate our approach on both non-scattering specimen (fluorescent beads suspended in agarose gel) and scattering media (brain tissues of genetically encoded mice). Extensive experiments demonstrate that our approach can reliably detect the 3D positions of granular targets with small Root Mean Square Error (RMSE), high robustness to optical aberration and light scattering in mammalian brain tissues.

Journal article

Howe CL, Quicke P, Song P, Jadan HV, Dragotti PL, Foust AJet al., 2020, Comparing wide field to 3d light field for imaging red calcium transients in mammalian brain

We apply light field (LF) microscopy to single-cell and bulk-loaded imaging of the red calcium dye, CaSiR-1 in mouse brain slices. We characterize the signal-to-noise ratio of images reconstructed from LF to wide-field time series.

Conference paper

Quicke P, Song C, McKimm EJ, Milosevic MM, Howe CL, Neil M, Schultz SR, Antic SD, Foust AJ, Knopfel Tet al., 2019, Corrigendum: Single-neuron level one-photon voltage imaging with sparsely targeted genetically encoded voltage indicators, Frontiers in Cellular Neuroscience, Vol: 13, ISSN: 1662-5102

Voltage imaging of many neurons simultaneously at single-cell resolution is hampered bythe difficulty of detecting small voltage signals from overlapping neuronal processes inneural tissue. Recent advances in genetically encoded voltage indicator (GEVI) imaginghave shown single-cell resolution optical voltage recordings in intact tissue throughimaging naturally sparse cell classes, sparse viral expression, soma restricted expression,advanced optical systems, or a combination of these. Widespread sparse and strongtransgenic GEVI expression would enable straightforward optical access to a denselyoccurring cell type, such as cortical pyramidal cells. Here we demonstrate that a recentlydescribed sparse transgenic expression strategy can enable single-cell resolution voltageimaging of cortical pyramidal cells in intact brain tissue without restricting expression tothe soma. We also quantify the functional crosstalk in brain tissue and discuss optimalimaging rates to inform future GEVI experimental design.

Journal article

Soor N, Quicke P, Howe C, Pang KT, Neil M, Schultz S, Foust Aet al., 2019, All-optical crosstalk-free manipulation and readout of Chronos-expressing neurons, Journal of Physics D: Applied Physics, Vol: 52, Pages: 1-10, ISSN: 0022-3727

All optical neurophysiology allows manipulation and readout of neural network activity with single-cell spatial resolution and millisecond temporal resolution. Neurons can be made to express proteins that actuate transmembrane currents upon light absorption, enabling optical control of membrane potential and action potential signalling. In addition, neurons can be genetically or synthetically labelled with fluorescent reporters of changes in intracellular calcium concentration or membrane potential. Thus, to optically manipulate and readout neural activity in parallel, two spectra are involved: the action spectrum of the actuator, and the absorption spectrum of the fluorescent reporter. Due to overlap in these spectra, previous all-optical neurophysiology paradigms have been hindered by spurious activation of neuronal activity caused by the readout light. Here, we pair the blue-green absorbing optogenetic actuator, Chronos, with a deep red-emitting fluorescent calcium reporter CaSiR-1. We show that cultured Chinese hamster ovary cells transfected with Chronos do not exhibit transmembrane currents when illuminated with wavelengths and intensities suitable for exciting one-photon CaSiR-1 fluorescence. We then demonstrate crosstalk-free, high signal-to-noise ratio CaSiR-1 red fluorescence imaging at 100 frames s−1 of Chronos-mediated calcium transients evoked in neurons with blue light pulses at rates up to 20 Hz. These results indicate that the spectral separation between red light excited fluorophores, excited efficiently at or above 640 nm, with blue-green absorbing opsins such as Chronos, is sufficient to avoid spurious opsin actuation by the imaging wavelengths and therefore enable crosstalk-free all-optical neuronal manipulation and readout.

Journal article

Quicke P, Song C, McKimm EJ, Milosevic MM, Howe CL, Neil M, Schultz SR, Antic SD, Foust AJ, Knopfel Tet al., 2019, Single-neuron level one-photon voltage imaging with sparsely targeted genetically encoded voltage indicators, Frontiers in Cellular Neuroscience, Vol: 13, ISSN: 1662-5102

Voltage imaging of many neurons simultaneously at single-cell resolution is hampered by the difficulty of detecting small voltage signals from overlapping neuronal processes in neural tissue. Recent advances in genetically encoded voltage indicator (GEVI) imaging have shown single-cell resolution optical voltage recordings in intact tissue through imaging naturally sparse cell classes, sparse viral expression, soma restricted expression, advanced optical systems, or a combination of these. Widespread sparse and strong transgenic GEVI expression would enable straightforward optical access to a densely occurring cell type, such as cortical pyramidal cells. Here we demonstrate that a recently described sparse transgenic expression strategy can enable single-cell resolution voltage imaging of cortical pyramidal cells in intact brain tissue without restricting expression to the soma. We also quantify the functional crosstalk in brain tissue and discuss optimal imaging rates to inform future GEVI experimental design.

Journal article

Quicke P, Howe CL, Song P, Jadan HV, Dragotti PL, Knöpfel T, Foust AJ, Schultz SR, Neil Met al., 2019, Calculation of high numerical aperture lightfield microscope point spread functions

3D deconvolution of lightfield images enables high resolution reconstruction of sample volumes. Previous point spread function calculations assume low to moderate NA objectives. Here we present a simple vectorial calculation valid for high NA objectives.

Conference paper

Song P, Jadan HV, Quicke P, Howe CL, Foust AJ, Dragotti PLet al., 2019, Location estimation for light field microscopy based on convolutional sparse coding

In this work, we propose an algorithm to estimate the depth location of objects from lightfield microscopy data by leveraging the sparsity of Epipolar Plane Images (EPIs) and convolutional sparse coding.

Conference paper

Quicke P, Howe CL, Song P, Jadan HV, Dragotti PL, Knopfel T, Foust AJ, Schultz SR, Neil Met al., 2019, Calculation of high numerical aperture lightfield microscope point spread functions, Computational Optical Sensing and Imaging, Publisher: OSA

Conference paper

Quicke P, Reynolds S, Neil M, Knopfel T, Schultz S, Foust AJet al., 2018, High speed functional imaging with source localized multifocal two-photon microscopy, Biomedical Optics Express, Vol: 9, Pages: 3678-3693, ISSN: 2156-7085

Multifocal two-photon microscopy (MTPM) increases imaging speed over single-focus scanning by parallelizing fluorescence excitation. The imaged fluorescence’s susceptibility to crosstalk, however, severely degrades contrast in scattering tissue. Here we present a source-localized MTPM scheme optimized for high speed functional fluorescence imaging in scattering mammalian brain tissue. A rastered line array of beamlets excites fluorescence imaged with a complementary metal-oxide-semiconductor (CMOS) camera. We mitigate scattering-induced crosstalk by temporally oversampling the rastered image, generating grouped images with structured illumination, and applying Richardson-Lucy deconvolution to reassign scattered photons. Single images are then retrieved with a maximum intensity projection through the deconvolved image groups. This method increased image contrast at depths up to 112 μm in scattering brain tissue and reduced functional crosstalk between pixels during neuronal calcium imaging. Source-localization did not affect signal-to-noise ratio (SNR) in densely labeled tissue under our experimental conditions. SNR decreased at low frame rates in sparsely labeled tissue, with no effect at frame rates above 50 Hz. Our non-descanned source-localized MTPM system enables high SNR, 100 Hz capture of fluorescence transients in scattering brain, increasing the scope of MTPM to faster and smaller functional signals.

Journal article

Quicke P, Neil M, Knopfel T, Schultz SR, Foust AJet al., 2017, Source-Localized Multifocal Two-Photon Microscopy for High-Speed Functional Imaging, 71st Annual Meeting of the Society-of-General-Physiologists (SGP) on Optical Revolution in Physiology - From Membrane to Brain, Publisher: ROCKEFELLER UNIV PRESS, Pages: 13A-14A, ISSN: 0022-1295

Conference paper

Quicke P, Barnes SJ, Knöpfel T, 2017, Imaging of Brain Slices with a Genetically Encoded Voltage Indicator., Methods Mol Biol, Vol: 1563, Pages: 73-84

Functional fluorescence microscopy of brain slices using voltage sensitive fluorescent proteins (VSFPs) allows large scale electrophysiological monitoring of neuronal excitation and inhibition. We describe the equipment and techniques needed to successfully record functional responses optical voltage signals from cells expressing a voltage indicator such as VSFP Butterfly 1.2. We also discuss the advantages of voltage imaging and the challenges it presents.

Journal article

Schultz SR, Copeland CS, Foust AJ, Quicke P, Schuck Ret al., 2016, Advances in two-photon scanning and scanless microscopy technologies for functional neural circuit imaging, Proceedings of the IEEE, Vol: 105, Pages: 139-157, ISSN: 0018-9219

Recent years have seen substantial developments in technology for imaging neural circuits, raising the prospect of large-scale imaging studies of neural populations involved in information processing, with the potential to lead to step changes in our understanding of brain function and dysfunction. In this paper, we will review some key recent advances: improved fluorophores for single-cell resolution functional neuroimaging using a two-photon microscope; improved approaches to the problem of scanning active circuits; and the prospect of scanless microscopes which overcome some of the bandwidth limitations of current imaging techniques. These advances in technology for experimental neuroscience have in themselves led to technical challenges, such as the need for the development of novel signal processing and data analysis tools in order to make the most of the new experimental tools. We review recent work in some active topics, such as region of interest segmentation algorithms capable of demixing overlapping signals, and new highly accurate algorithms for calcium transient detection. These advances motivate the development of new data analysis tools capable of dealing with spatial or spatiotemporal patterns of neural activity that scale well with pattern size.

Journal article

Schuck R, Quicke P, Hwang JK, Annecchino L, Schultz SRet al., 2015, Rapid three dimensional two photon neural population scanning, 37th Annual International IEEE EMBS Conference of the IEEE Engineering in Medicine and Biology Society, Publisher: IEEE, Pages: 5867-5870, ISSN: 1557-170X

Recording the activity of neural populationsat high sampling rates is a fundamental requirement forunderstanding computation in neural circuits. Two photonmicroscopy provides one promising approach towards this.However, neural circuits are three dimensional, and functionalimaging in two dimensions fails to capture the 3D natureof neural dynamics. Electrically tunable lenses (ETLs) providea simple and cheap method to extend laser scanningmicroscopy into the relatively unexploited third dimension.We have therefore incorporated them into our Adaptive SpiralScanning (SSA) algorithm, which calculates kinematicallyefficient scanning strategies using radially modulated spiralpaths. We characterised the response of the ETL, incorporatedits dynamics using MATLAB models of the SSA algorithmand tested the models on populations of Izhikevich neuronsof varying size and density. From this, we show that ouralgorithms can theoretically at least achieve sampling rates of36.2Hz compared to 21.6Hz previously reported for 3D scanningtechniques.

Conference paper

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