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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 孫啟光(Chi-Kuang Sun) | |
dc.contributor.author | Han-Wee Chong | en |
dc.contributor.author | 張漢維 | zh_TW |
dc.date.accessioned | 2021-06-07T17:33:34Z | - |
dc.date.issued | 2020 | |
dc.date.submitted | 2021-03-26 | |
dc.identifier.citation | [1] C. C. Petersen, 'Whole-cell recording of neuronal membrane potential during behavior,' Neuron, vol. 95, no. 6, pp. 1266-1281, 2017.
[2] A. K. Lee and M. Brecht, 'Elucidating neuronal mechanisms using intracellular recordings during behavior,' Trends in neurosciences, vol. 41, no. 6, pp. 385-403, 2018. [3] D. Mishra, A. Yadav, S. Ray, and P. Kalra, 'Exploring biological neuron models,' Directions, The Research Magazine of IIT Kanpur, vol. 7, no. 3, pp. 13-22, 2006. [4] M. Lasser, J. Tiber, and L. A. Lowery, 'The role of the microtubule cytoskeleton in neurodevelopmental disorders,' Frontiers in cellular neuroscience, vol. 12, p. 165, 2018. [5] J. F. Fulton, 'Physiology of the nervous system, rev,' 1938. [6] D. S. Faber and A. E. Pereda, 'Two forms of electrical transmission between neurons,' Frontiers in Molecular Neuroscience, vol. 11, p. 427, 2018. [7] E. M. Izhikevich and J. Moehlis, 'Dynamical Systems in Neuroscience: The geometry of excitability and bursting,' SIAM review, vol. 50, no. 2, p. 397, 2008. [8] R. Plonsey and R. C. Barr, Bioelectricity: a quantitative approach. Springer Science Business Media, 2007. [9] J. Juchem, 'Development of a mathematical model, method and device for Continuous Monitoring of Skin Impedance for Analgesia,' 2017. [10] R. A. Capel and D. A. Terrar, 'The importance of Ca2+-dependent mechanisms for the initiation of the heartbeat,' Frontiers in physiology, vol. 6, p. 80, 2015. [11] W. Gerstner and W. M. Kistler, Spiking neuron models: Single neurons, populations, plasticity. Cambridge university press, 2002. [12] J. Bodurka and P. A. Bandettini, 'Toward direct mapping of neuronal activity: MRI detection of ultraweak, transient magnetic field changes,' Magnetic Resonance in Medicine: An Official Journal of the International Society for Magnetic Resonance in Medicine, vol. 47, no. 6, pp. 1052-1058, 2002. [13] J. A. Detre, J. Wang, Z. Wang, and H. Rao, 'Arterial spin-labeled perfusion MRI in basic and clinical neuroscience,' Current opinion in neurology, vol. 22, no. 4, pp. 348-355, 2009. [14] J. A. Detre, 'Clinical applicability of functional MRI,' Journal of Magnetic Resonance Imaging: An Official Journal of the International Society for Magnetic Resonance in Medicine, vol. 23, no. 6, pp. 808-815, 2006. [15] E. Neher and B. Sakmann, 'Single-channel currents recorded from membrane of denervated frog muscle fibres,' Nature, vol. 260, no. 5554, pp. 799-802, 1976. [16] E. Neher, '[6] Correction for liquid junction potentials in patch clamp experiments,' Methods in enzymology, vol. 207, pp. 123-131, 1992. [17] B. Sakmann and E. Neher, 'Patch clamp techniques for studying ionic channels in excitable membranes,' Annual review of physiology, vol. 46, no. 1, pp. 455-472, 1984. [18] O. P. Hamill, A. Marty, E. Neher, B. Sakmann, and F. Sigworth, 'Improved patch-clamp techniques for high-resolution current recording from cells and cell-free membrane patches,' Pflügers Archiv, vol. 391, no. 2, pp. 85-100, 1981. [19] A. L. Hodgkin and A. F. Huxley, 'A quantitative description of membrane current and its application to conduction and excitation in nerve,' The Journal of physiology, vol. 117, no. 4, p. 500, 1952. [20] Z. Zhu, R. Wang, and F. Zhu, 'The energy coding of a structural neural network based on the Hodgkin–Huxley model,' Frontiers in neuroscience, vol. 12, p. 122, 2018. [21] J. Guckenheimer and R. A. Oliva, 'Chaos in the Hodgkin--Huxley Model,' SIAM Journal on Applied Dynamical Systems, vol. 1, no. 1, pp. 105-114, 2002. [22] M. Z. Lin and M. J. Schnitzer, 'Genetically encoded indicators of neuronal activity,' Nature neuroscience, vol. 19, no. 9, p. 1142, 2016. [23] G. Buzsáki, 'Large-scale recording of neuronal ensembles,' Nature neuroscience, vol. 7, no. 5, pp. 446-451, 2004. [24] A. R. Kay and R. K. Wong, 'Isolation of neurons suitable for patch-clamping from adult mammalian central nervous systems,' Journal of neuroscience methods, vol. 16, no. 3, pp. 227-238, 1986. [25] K.-J. Halbhuber and K. König, 'Modern laser scanning microscopy in biology, biotechnology and medicine,' Annals of Anatomy-Anatomischer Anzeiger, vol. 185, no. 1, pp. 1-20, 2003. [26] S. J. Wright and D. J. Wright, 'Introduction to confocal microscopy,' Cell Biological Applications of Confocal Microscopy, in Methods in Cell Biology, vol. 70, pp. 1-85, 2002. [27] F. Helmchen and W. Denk, 'New developments in multiphoton microscopy,' Current opinion in neurobiology, vol. 12, no. 5, pp. 593-601, 2002. [28] D. R. Larson et al., 'Water-soluble quantum dots for multiphoton fluorescence imaging in vivo,' Science, vol. 300, no. 5624, pp. 1434-1436, 2003. [29] M. D. Cahalan, I. Parker, S. H. Wei, and M. J. Miller, 'Real-time imaging of lymphocytes in vivo,' Current opinion in immunology, vol. 15, no. 4, pp. 372-377, 2003. [30] P. Bousso and E. A. Robey, 'Dynamic behavior of T cells and thymocytes in lymphoid organs as revealed by two-photon microscopy,' Immunity, vol. 21, no. 3, pp. 349-355, 2004. [31] B. A. Molitoris and R. M. Sandoval, 'Intravital multiphoton microscopy of dynamic renal processes,' American Journal of Physiology-Renal Physiology, vol. 288, no. 6, pp. F1084-F1089, 2005. [32] M. Rubart, 'Two-photon microscopy of cells and tissue,' Circulation research, vol. 95, no. 12, pp. 1154-1166, 2004. [33] L. L. Hsu, S. B. Pelet, T. M. Hancewicz, P. D. Kaplan, and P. T. So, 'Two-photon 3-D mapping of ex vivo human skin endogenous fluorescence species based on fluorescence emission spectra,' Journal of biomedical optics, vol. 10, no. 2, p. 024016, 2005. [34] V. E. Centonze and J. G. White, 'Multiphoton excitation provides optical sections from deeper within scattering specimens than confocal imaging,' Biophysical journal, vol. 75, no. 4, pp. 2015-2024, 1998. [35] K. Svoboda and R. Yasuda, 'Principles of two-photon excitation microscopy and its applications to neuroscience,' Neuron, vol. 50, no. 6, pp. 823-839, 2006. [36] R. Prevedel et al., 'Simultaneous whole-animal 3D imaging of neuronal activity using light-field microscopy,' Nature methods, vol. 11, no. 7, pp. 727-730, 2014. [37] H. Dana et al., 'Sensitive red protein calcium indicators for imaging neural activity,' Elife, vol. 5, p. e12727, 2016. [38] Y. Ziv et al., 'Long-term dynamics of CA1 hippocampal place codes,' Nature neuroscience, vol. 16, no. 3, p. 264, 2013. [39] G. J. Broussard, R. Liang, and L. Tian, 'Monitoring activity in neural circuits with genetically encoded indicators,' (in English), Frontiers in Molecular Neuroscience, Review vol. 7, no. 97, 2014-December-05 2014, doi: 10.3389/fnmol.2014.00097. [40] A. Miyawaki et al., 'Fluorescent indicators for Ca 2+ based on green fluorescent proteins and calmodulin,' Nature, vol. 388, no. 6645, pp. 882-887, 1997. [41] D. W. Tank, M. Sugimori, J. A. Connor, and R. R. Llinas, 'Spatially resolved calcium dynamics of mammalian Purkinje cells in cerebellar slice,' Science, vol. 242, no. 4879, pp. 773-777, 1988. [42] C. Grienberger and A. Konnerth, 'Imaging calcium in neurons,' Neuron, vol. 73, no. 5, pp. 862-885, 2012. [43] L. Tian, J. Akerboom, E. R. Schreiter, and L. L. Looger, 'Neural activity imaging with genetically encoded calcium indicators,' in Progress in brain research, vol. 196: Elsevier, 2012, pp. 79-94. [44] M. Lin and M. Schnitzer, 'Genetically encoded indicators of neuronal activity. Nature6Neuroscience,' ed, 2016. [45] J. Nakai, M. Ohkura, and K. Imoto, 'A high signal-to-noise Ca 2+ probe composed of a single green fluorescent protein,' Nature biotechnology, vol. 19, no. 2, pp. 137-141, 2001. [46] T.-W. Chen et al., 'Ultrasensitive fluorescent proteins for imaging neuronal activity,' Nature, vol. 499, no. 7458, pp. 295-300, 2013. [47] R. Lu et al., 'Rapid mesoscale volumetric imaging of neural activity with synaptic resolution,' Nature Methods, vol. 17, no. 3, pp. 291-294, 2020. [48] S. Chamberland et al., 'Fast two-photon imaging of subcellular voltage dynamics in neuronal tissue with genetically encoded indicators,' Elife, vol. 6, Jul 27 2017, doi: 10.7554/eLife.25690. [49] T. Knöpfel, Y. Gallero-Salas, and C. Song, 'Genetically encoded voltage indicators for large scale cortical imaging come of age,' Current opinion in chemical biology, vol. 27, pp. 75-83, 2015. [50] H. H. Yang and F. St-Pierre, 'Genetically encoded voltage indicators: opportunities and challenges,' Journal of Neuroscience, vol. 36, no. 39, pp. 9977-9989, 2016. [51] W. Akemann, H. Mutoh, A. Perron, J. Rossier, and T. Knöpfel, 'Imaging brain electric signals with genetically targeted voltage-sensitive fluorescent proteins,' Nature methods, vol. 7, no. 8, pp. 643-649, 2010. [52] G. Scott et al., 'Voltage imaging of waking mouse cortex reveals emergence of critical neuronal dynamics,' Journal of Neuroscience, vol. 34, no. 50, pp. 16611-16620, 2014. [53] M. Carandini, D. Shimaoka, L. F. Rossi, T. K. Sato, A. Benucci, and T. Knöpfel, 'Imaging the awake visual cortex with a genetically encoded voltage indicator,' Journal of Neuroscience, vol. 35, no. 1, pp. 53-63, 2015. [54] H. Mutoh, Y. Mishina, Y. Gallero-Salas, and T. Knöpfel, 'Comparative performance of a genetically-encoded voltage indicator and a blue voltage sensitive dye for large scale cortical voltage imaging,' Frontiers in Cellular Neuroscience, vol. 9, p. 147, 2015. [55] D. Shimaoka, C. Song, and T. Knöpfel, 'State-dependent modulation of slow wave motifs towards awakening,' Frontiers in cellular neuroscience, vol. 11, p. 108, 2017. [56] K. F. Ahrens, B. Heider, H. Lee, E. Y. Isacoff, and R. M. Siegel, 'Two-photon scanning microscopy of in vivo sensory responses of cortical neurons genetically encoded with a fluorescent voltage sensor in rat,' Frontiers in neural circuits, vol. 6, p. 15, 2012. [57] N. C. Flytzanis et al., 'Archaerhodopsin variants with enhanced voltage-sensitive fluorescence in mammalian and Caenorhabditis elegans neurons,' Nature communications, vol. 5, no. 1, pp. 1-9, 2014. [58] Y. Gong, M. Wagner, J. Zhong Li, and M. Schnitzer, 'Imaging neural spiking in brain tissue using FRET-opsin protein voltage sensors. Nat Commun 5: 3674,' ed, 2014. [59] Y. Gong et al., 'High-speed recording of neural spikes in awake mice and flies with a fluorescent voltage sensor,' Science, vol. 350, no. 6266, pp. 1361-1366, 2015. [60] W. Denk, J. H. Strickler, and W. W. Webb, 'Two-photon laser scanning fluorescence microscopy,' Science, vol. 248, no. 4951, pp. 73-76, 1990. [61] S. W. Perry, R. M. Burke, and E. B. Brown, 'Two-photon and second harmonic microscopy in clinical and translational cancer research,' Annals of biomedical engineering, vol. 40, no. 2, pp. 277-291, 2012. [62] P. T. So, C. Y. Dong, B. R. Masters, and K. M. Berland, 'Two-photon excitation fluorescence microscopy,' Annual review of biomedical engineering, vol. 2, no. 1, pp. 399-429, 2000. [63] W. Denk, D. W. Piston, and W. W. Webb, 'Two-photon molecular excitation in laser-scanning microscopy,' in Handbook of biological confocal microscopy: Springer, 1995, pp. 445-458. [64] W. Denk and K. Svoboda, 'Photon upmanship: why multiphoton imaging is more than a gimmick,' Neuron, vol. 18, no. 3, pp. 351-357, 1997. [65] W. R. Zipfel, R. M. Williams, and W. W. Webb, 'Nonlinear magic: multiphoton microscopy in the biosciences,' Nature biotechnology, vol. 21, no. 11, pp. 1369-1377, 2003. [66] P. J. Campagnola and L. M. Loew, 'Second-harmonic imaging microscopy for visualizing biomolecular arrays in cells, tissues and organisms,' Nature biotechnology, vol. 21, no. 11, pp. 1356-1360, 2003. [67] J. Mertz, 'Nonlinear microscopy: new techniques and applications,' Current opinion in neurobiology, vol. 14, no. 5, pp. 610-616, 2004. [68] D. S. Peterka, H. Takahashi, and R. Yuste, 'Imaging voltage in neurons,' Neuron, vol. 69, no. 1, pp. 9-21, 2011. [69] A. Sobczyk, V. Scheuss, and K. Svoboda, 'NMDA receptor subunit-dependent [Ca2+] signaling in individual hippocampal dendritic spines,' Journal of Neuroscience, vol. 25, no. 26, pp. 6037-6046, 2005. [70] J. Hao and T. G. Oertner, 'Depolarization gates spine calcium transients and spike-timing-dependent potentiation,' Current opinion in neurobiology, vol. 22, no. 3, pp. 509-515, 2012. [71] K. P. Lillis, A. Eng, J. A. White, and J. Mertz, 'Two-photon imaging of spatially extended neuronal network dynamics with high temporal resolution,' Journal of neuroscience methods, vol. 172, no. 2, pp. 178-184, 2008. [72] T. Zhang et al., 'Kilohertz two-photon brain imaging in awake mice,' Nature methods, vol. 16, no. 11, pp. 1119-1122, 2019. [73] Y. Adam et al., 'Voltage imaging and optogenetics reveal behaviour-dependent changes in hippocampal dynamics,' Nature, vol. 569, no. 7756, pp. 413-417, 2019. [74] G. Fan, H. Fujisaki, A. Miyawaki, R.-K. Tsay, R. Y. Tsien, and M. H. Ellisman, 'Video-rate scanning two-photon excitation fluorescence microscopy and ratio imaging with cameleons,' Biophysical journal, vol. 76, no. 5, pp. 2412-2420, 1999. [75] D. A. Storace, O. R. Braubach, L. Jin, L. B. Cohen, and U. Sung, 'Monitoring brain activity with protein voltage and calcium sensors,' Scientific reports, vol. 5, p. 10212, 2015. [76] H. H. Yang, F. St-Pierre, X. Sun, X. Ding, M. Z. Lin, and T. R. Clandinin, 'Subcellular imaging of voltage and calcium signals reveals neural processing in vivo,' Cell, vol. 166, no. 1, pp. 245-257, 2016. [77] M. B. Ahrens, M. B. Orger, D. N. Robson, J. M. Li, and P. J. Keller, 'Whole-brain functional imaging at cellular resolution using light-sheet microscopy,' Nature methods, vol. 10, no. 5, pp. 413-420, 2013. [78] W. Akemann, M. Sasaki, H. Mutoh, T. Imamura, N. Honkura, and T. Knöpfel, 'Two-photon voltage imaging using a genetically encoded voltage indicator,' Scientific reports, vol. 3, p. 2231, 2013. [79] B. Baker et al., 'Three fluorescent protein voltage sensors exhibit low plasma membrane expression in mammalian cells,' Journal of neuroscience methods, vol. 161, no. 1, pp. 32-38, 2007. [80] R. L. a. L. T. Gerard J. Broussard, 'Monitoring activity in neural circuits with genetically encoded indicators,' frontiers in molecular neuroscience, 2014. [81] S. Lou et al., 'Genetically targeted all-optical electrophysiology with a transgenic Cre-dependent optopatch mouse,' Journal of Neuroscience, vol. 36, no. 43, pp. 11059-11073, 2016. [82] V. Villette et al., 'Ultrafast two-photon imaging of a high-gain voltage indicator in awake behaving mice,' Cell, vol. 179, no. 7, pp. 1590-1608. e23, 2019. [83] F. St-Pierre, J. D. Marshall, Y. Yang, Y. Gong, M. J. Schnitzer, and M. Z. Lin, 'High-fidelity optical reporting of neuronal electrical activity with an ultrafast fluorescent voltage sensor,' Nature neuroscience, vol. 17, no. 6, pp. 884-889, 2014. [84] M. N. Modi, K. Daie, G. C. Turner, and K. Podgorski, 'Two-photon imaging with silicon photomultipliers,' Optics Express, vol. 27, no. 24, pp. 35830-35841, 2019. [85] W. P. Chan and M. Je, 'A review of CMOS multimodal neuromonitoring sensors and systems,' in 2011 International Symposium on Integrated Circuits, 2011: IEEE, pp. 416-419. [86] M. Abdallah and O. Elkeelany, 'A survey on data acquisition systems DAQ,' in 2009 International Conference on Computing, Engineering and Information, 2009: IEEE, pp. 240-243. [87] B. Haji-Saeed, J. Khoury, C. L. Woods, D. Pyburn, S. K. Sengupta, and J. Kierstead, 'Mapping approach for image correction and processing for bidirectional resonant scanners,' Optical Engineering, vol. 46, no. 2, p. 027007, 2007. [88] B. Hangün and Ö. Eyecioğlu, 'Performance comparison between OpenCV built in CPU and GPU functions on image processing operations,' arXiv preprint arXiv:1906.08819, 2019. [89] S. Kameyama and Y. Miura, 'Research for High Speed Image Processing Programming Method on Combined Environment of CUDA and OpenCV,' in 2018 IEEE International Conference on Consumer Electronics-Taiwan (ICCE-TW), 2018: IEEE, pp. 1-2. [90] P. Garg and T. Jain, 'A comparative study on histogram equalization and cumulative histogram equalization,' International Journal of New Technology and Research, vol. 3, no. 9, 2017. [91] P. Merken and R. Vandersmissen, 'Dark current and influence of target emissivity,' Photonics Imaging Technol, 2016. [92] P. Tubes, 'Basics and Applications, Hamamatsu Photonics (2007) www. hamamatsu. com/resources/pdf/etd/PMT_handbook_v3aE. pdf,' Accessed on May 3rd, 2017. [93] U. Erkan, L. Gökrem, and S. Enginoğlu, 'Different applied median filter in salt and pepper noise,' Computers Electrical Engineering, vol. 70, pp. 789-798, 2018. [94] K. K. V. Toh, H. Ibrahim, and M. N. Mahyuddin, 'Salt-and-pepper noise detection and reduction using fuzzy switching median filter,' IEEE Transactions on Consumer Electronics, vol. 54, no. 4, pp. 1956-1961, 2008. [95] S. Esakkirajan, T. Veerakumar, A. N. Subramanyam, and C. PremChand, 'Removal of high density salt and pepper noise through modified decision based unsymmetric trimmed median filter,' IEEE Signal processing letters, vol. 18, no. 5, pp. 287-290, 2011. [96] R. Lu et al., 'Rapid mesoscale volumetric imaging of neural activity with synaptic resolution,' Nat Methods, vol. 17, no. 3, pp. 291-294, Mar 2020, doi: 10.1038/s41592-020-0760-9. [97] J. Wu et al., 'Kilohertz two-photon fluorescence microscopy imaging of neural activity in vivo,' Nature Methods, vol. 17, no. 3, pp. 287-290, 2020. [98] T. Komiyama et al., 'Learning-related fine-scale specificity imaged in motor cortex circuits of behaving mice,' Nature, vol. 464, no. 7292, pp. 1182-1186, 2010. [99] K. B. Hengen, M. E. Lambo, S. D. Van Hooser, D. B. Katz, and G. G. Turrigiano, 'Firing rate homeostasis in visual cortex of freely behaving rodents,' Neuron, vol. 80, no. 2, pp. 335-342, 2013. [100] J.-A. Conchello et al., 'A video rate laser scanning confocal microscope,' presented at the Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XV, 2008. [101] M. Gardner and D. Altman, 'Calculating confidence intervals for proportions and their differences,' Statistics with confidence. London: BMJ Publishing Group, pp. 28-33, 1989. [102] H. C. Burger, C. J. Schuler, and S. Harmeling, 'Image denoising: Can plain neural networks compete with BM3D?,' in 2012 IEEE conference on computer vision and pattern recognition, 2012: IEEE, pp. 2392-2399. [103] M. Chu, Y. Xie, J. Mayer, L. Leal-Taixé, and N. Thuerey, 'Learning temporal coherence via self-supervision for GAN-based video generation,' ACM Transactions on Graphics (TOG), vol. 39, no. 4, pp. 75: 1-75: 13, 2020. | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/15383 | - |
dc.description.abstract | 神經元之間通過動作電位交互作用,其所耗之時間往往發生在毫秒這個級距內。因此,我們需要可以提供高時間解析度的光學顯微鏡,用以研究神經元細胞之間的神經迴路。為此,我們需要具有上千赫茲掃描功能的雙光子熒光顯微鏡(2PFM),用以監控動作電位來描述神經元之間的電壓變化。此外,利用Accelerated Sensor of Action Potentials (ASAPs)與2PFM的結合,可以從大量清醒細胞中進行無創、高靈敏度和長時間的光學記錄;這將為理解神經系統如何在神經迴路和神經訊息編碼和處理訊息鋪平道路。本篇論文的重點將描述兩千赫茲分辨率的電壓影像。
本文中,為了進行原理驗證研究,我們首先演示我們的超高速2PFM數位電控成果實現及其特性。其次,我們也分享了毫秒級精度的ASAPs雙光子電壓影像。在這裡,利用我們擁有2k幀率顯微鏡的優勢,我們將其用以觀測清醒小鼠大腦中的神經元細胞。最後,以2k幀率的ASAPs結果,我們已經能夠獲得其動作電位。高時間解析度不僅提供了動作電位的更多細節,而且還提供了更高精準度。 使用這個平台,可以觀測到體內的動作電位。而且這超高時間解析度的研究,至今沒有學者公開發表過。我們的系統非常穩定、強大、完整、以及具有極大的成本效益。這平台為研究動物行為打下了強大的可能性。將來,不僅是神經元迴路的病理分析,這平台還可用以不同的生醫影像研究。 | zh_TW |
dc.description.abstract | The neuronal communication through action potential and neurotransmitter release occurs on millisecond time scale. Thus, we need optical microscopy tools which can provide high temporal resolution to investigate activity of neuron cells and consequently, neuronal circuits. For this, we need ultrahigh speed sub-resolution optical imaging tools viz. two-photon fluoresce microscopy (2PFM) with kilohertz frequency scanning capability to monitor the action potential signal, depicting the voltage signaling in neurons. Consequently, combining the imaging of voltage sensor, Accelerated Sensor of Action Potentials (ASAPs) with high-speed 2PFM, allows non-invasive, ultrasensitive and chronic simultaneous optical recording from large populations of awake cells; which will pave the way in understanding the how nervous systems encode and process information at circuit and single-cell levels. However, the technological development in this regard is still to report the voltage imaging at kilohertz sub-resolution.
In this thesis, for a proof-of-principle study, we, for the first time, will demonstrate our digital architecture implementation of Ultra-high speed 2PFM and its characteristics. Second, we will demonstrate ASAPs-based two-photon voltage imaging with millisecond-timescale precision. Here, taking the advantages of our microscopy with nearly 2k frame rate high-speed scanning capability, we will apply it on neuron cells observation in awake mice brain, particularly for visual cortical neurons. Finally, taking the 2k frame rate ASAPs results, we have been able to obtain its action potentials. The high temporal resolution of neuron observation not only provides more details of action potential, but also higher precision of action potential recording. With this platform, in vivo measurement for action potential could be thus observed, with time-resolution not yet reported. Our system is significantly robust, stable, compact, and cost-effective with comparable spatial resolution. This platform opens the possibility for detailed explorations of network dynamics in the context of behavior in animals. In the future, not only pathological features on neuronal circuit, the developed imaging platform will be further utilized for varied other investigations of biomedical importance. | en |
dc.description.provenance | Made available in DSpace on 2021-06-07T17:33:34Z (GMT). No. of bitstreams: 1 U0001-0202202116254000.pdf: 18910529 bytes, checksum: 7da6dedf27f7ec561b6f146c14cc22ab (MD5) Previous issue date: 2020 | en |
dc.description.tableofcontents | CHAPTER 1. INTRODUCTION 1 1.1 Neurons and action potential 1 1.2 Traditional methods to monitor action potential 4 1.3 Optical methods for studying neural activity 6 1.3.1 Genetically Encoded indicators of Neural Activity, GINAs 7 CHAPTER 2. THESIS MOTIVATION AND OBJECTIVES 10 2.1 Digital architecture implementation for an Ultra-high speed Two-photon fluorescence microscopy system 10 2.2 Accelerated Sensor of Action Potentials (ASAP) 12 2.3 Measurement of action potential 12 CHAPTER 3. MATERIAL AND METHODS 14 3.1 Specimen preparation 14 3.1.1 Mice 14 3.1.2 Preparation of C57BL/6-Tg Thy-1 EGFP mouse brain slices 14 3.1.3 Injection of voltage indicator, ASAP3 into mice brain 15 3.1.4 Development of cranial window to observe the ASAP3 expressing neuron in vivo 15 3.2 Optical Setup 17 CHAPTER 4. Digital architecture implementation and software 19 4.1 Introduction 19 4.2 Scanning System 21 4.2.1 Resonant Scanner 21 4.2.2 Galvanometer Scanner 23 4.3 Signal Acquisition System 25 4.3.1 Field-Programmable Gate Array (FPGA) 27 4.3.2 FPGA program architecture 28 4.3.3 Serial data communication 30 4.4 Processing System 32 4.4.1 Host-PC hardware setup 32 4.4.2 Real-time image display 34 4.4.3 Image Analysis 39 4.5 Storage System 39 4.6 UFO Microscopy whole structure 41 CHAPTER 5. RESULTS 42 5.1 System characterizations 44 5.1.1 Image Processing Results 44 5.1.2 FOV of x-axis and fix distortion comparison 44 5.1.3 FOV of Y-axis 48 5.1.4 Thy-1 EGFP transgenic mice brain tissue image 49 5.2 ASAP 50 5.2.1 Photodamage 50 5.2.2 Voltage imaging 50 5.2.3 Measuring the Action potential of neurons expressing ASAP3 as voltage indicator 51 CHAPTER 6. DISCUSSIONS 62 CHAPTER 7. CONCLUSIONS 65 CHAPTER 8. REFERENCES 66 Appendix 1: Licenses 72 | |
dc.language.iso | en | |
dc.title | 超高速雙光子顯微鏡之電控及其應用 | zh_TW |
dc.title | Digital architecture implementation of Ultra-high speed Two-photon fluorescence microscopy | en |
dc.type | Thesis | |
dc.date.schoolyear | 109-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 陳示國(Shih-Kuo Chen),宋孔彬(Kung-Bin Sung) | |
dc.subject.keyword | 雙光子熒光顯微鏡,超高速,生醫影像,動作電位, | zh_TW |
dc.subject.keyword | 2PFM,Ultra-high speed,Voltage imaging,ASAPs,Action Potentials, | en |
dc.relation.page | 82 | |
dc.identifier.doi | 10.6342/NTU202100397 | |
dc.rights.note | 未授權 | |
dc.date.accepted | 2021-03-26 | |
dc.contributor.author-college | 電機資訊學院 | zh_TW |
dc.contributor.author-dept | 光電工程學研究所 | zh_TW |
顯示於系所單位: | 光電工程學研究所 |
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