Multi-area two-photon microscopy for revealing long-distance communication between multiple local brain circuits

多区域双光子显微镜揭示多个局部脑回路之间的长距离通信

• 批准号: 9128075
• 负责人: Fritjof Helmchen
• 金额: $ 21.63万
• 依托单位: UNIVERSITY OF ZURICH
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-30 至 2017-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9128075
• 关键词: Accelerometer; Address; Animals; Area; Behavior; Behavioral; Biological; Brain; Brain region; Calcium; Caliber; Cells; Communication; Communities; Complex; Computer software; Custom; Dendrites; Detection; Discrimination; Distant; Ensure; Fiber; Fluorescence; Functional Imaging; Generations; Goals; Head; Health; Image; Image Analysis; Imaging Device; Imaging technology; Individual; Label; Lasers; Life; Measurement; Measures; Methods; Microscope; Microscopy; Monitor; Mus; Neocortex; Neurons; Neurosciences; Operating System; Optics; Pattern; Performance; Population; Population Dynamics; Positioning Attribute; Process; Property; Research; Resolution; Rodent; Scanning; Sensory; Side; Signal Transduction; Somatosensory Cortex; Speed; Support System; Synapses; System; Tactile; Technology; Testing; Texture; Tissues; Vibrissae; Viral; Work; awake; base; calcium indicator; cell type; cellular imaging; design; experience; fluorescence microscope; imaging modality; improved; in vivo; in vivo imaging; information processing; innovation; instrument; interest; neocortical; neural circuit; neuronal cell body; novel; optical imaging; prototype; relating to nervous system; research study; somatosensory; spatiotemporal; temporal measurement; two-photon

 DESCRIPTION (provided by applicant): Two-photon microscopy is a widely used, key method for functional imaging of cellular activity in living animals. Most recently, in vivo calciu imaging experiments have started to reveal the spatiotemporal activity patterns that occur in various areas of the neocortex during head-fixed mouse behavior. Typically, however, the field-of- view for imaging cellular activity is fairly small, on the order of a few hundred micrometers. This restriction limits the size of neuronal networks that can be studied and thus leaves open the question how local neuronal networks communicate with distant, synaptically connected regions. What is therefore needed is a new imaging instrument that allows high-resolution functional imaging from two regions simultaneously, in the best case identifying the mutual projection neurons. We have developed a novel multi-area 2-photon microscope (MA2PM) that fulfills this need by enabling simultaneous imaging of two sub-areas within a large global scan- field (1.8 mm diameter, a distance equivalent to lining 100,000 neuronal cell bodies side-by-side assuming 20 um per neuron). Such two areas can be independently and flexibly positioned, making it for example possible to simultaneously measure neurons in the primary and secondary somatosensory areas of mouse neocortex (> 1 mm apart) while the mouse is performing a tactile texture discrimination task with its whiskers. The Using the MA2PM prototype we have conducted first proof-of-principle experiments in somatosensory cortex of awake, behaving mice using a genetically-encoded calcium indicator. We apply viral retrograde labeling strategies to identify the subsets of neurons that give rise to the inter-areal connection. Our goal in the proposed project is to further optimize and extend this innovative, transforming microscopy technology in several ways. Aim 1: We will work on finalizing a full microscope design for 2-area imaging and demonstrate its usefulness for research on corticocortical processing. We are fully dedicated to build a modular, carefully designed instrument, perform a quantitative system characterization, and disseminate this new instrument to the broader neuroscience community, especially to the growing number of research groups applying two-photon imaging for the analysis of cortical processing. Aim 2: We will furthermore expand the system to an even larger field-of-view and extend it for simultaneous imaging of four sub-areas. To this end we will apply newest laser technology and employ state-of-the-art red-shifted genetically encoded calcium indicator for deep imaging. We particularly aim at instantiating two-layer imaging (e.g. in layers L2/3 and L5) in two connected cortical areas. Overall, we expect that the new multi-area imaging technology will be an enabling technology to bridge the level of local microcircuits to the 'macrocircuit' level of communicating brain areas and thus will be of immediate and broad interest and highest significance to the neuroscience community.



项目成果

Reflective multi-immersion microscope objectives inspired by the Schmidt telescope.

• DOI: 10.1038/s41587-023-01717-8
• 发表时间: 2024-01
• 期刊: NATURE BIOTECHNOLOGY
• 影响因子: 46.9
• 作者: Voigt, Fabian F. F.;Reuss, Anna Maria;Naert, Thomas;Hildebrand, Sven;Schaettin, Martina;Hotz, Adriana L. L.;Whitehead, Lachlan;Bahl, Armin;Neuhauss, Stephan C. F.;Roebroeck, Alard;Stoeckli, Esther T. T.;Lienkamp, Soeren S. S.;Aguzzi, Adriano;Helmchen, Fritjof
• 通讯作者: Helmchen, Fritjof

Multiphoton in vivo imaging with a femtosecond semiconductor disk laser.

• DOI: 10.1364/boe.8.003213
• 发表时间: 2017-07
• 期刊: Biomedical optics express
• 影响因子: 3.4
• 作者: F. Voigt;F. Emaury;P. Bethge;D. Waldburger;S. Link;Stefano Carta;Alexander van der Bourg;F. Helmchen;U. Keller

Long-range population dynamics of anatomically defined neocortical networks

• DOI: 10.7554/elife.14679
• 发表时间: 2016-05-24
• 期刊: ELIFE
• 影响因子: 7.7
• 作者: Chen, Jerry L.;Voigt, Fabian F.;Helmchen, Fritjof
• 通讯作者: Helmchen, Fritjof