Northern Lights collaboration for better 2-photon probes

北极光合作打造更好的 2 光子探测器

• 批准号: 9336980
• 负责人: Robert E. Campbell
• 金额: $ 64.46万
• 依托单位: MONTANA STATE UNIVERSITY - BOZEMAN
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-30 至 2021-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9336980
• 关键词: Amplifiers; BRAIN initiative; Benchmarking; Biosensor; Brain; Canada; Cells; Collaborations; Complex; Coupled; Data; Development; Engineering; Equipment; Escherichia coli; Evolution; Experimental Designs; Fluorescent Probes; Image; Laboratories; Lasers; Measurement; Methodology; Microscope; Microscopy; Molecular; Montana; Nature; Neurons; Persons; Photons; Physiologic pulse; Preparation; Process; Property; Protein Engineering; Proteins; Research Personnel; Resolution; Sampling; Signal Transduction; Specificity; Spectrum Analysis; System; Technology; Testing; Thick; Time; Tissue imaging; Tissues; Training; Zebrafish; absorption; base; cellular imaging; experimental study; instrument; interest; multi-photon; mutant; neural circuit; neuronal circuitry; next generation; novel; programs; public health relevance; red fluorescent protein; regenerative; relating to nervous system; sapphire laser; screening; sensor; skills; tool; two-photon; voltage

DESCRIPTION (provided by applicant): The Northern Lights collaboration for better 2-photon probes will combine the expertise of four very different investigators and laboratories from Montana and Canada with complementary interests and skills to tackle a fundamental problem in imaging of the activity of the living brain. It is now well agreed that genetically encoded, fluorescent biosensors are uniquely suited to provide the specificity and versatility necessary to image the activity of defined cells in the brain. It is also clear that to image cells in thick slies or whole brains it will be necessary to turn to 2- photon (nonlinear) imaging, which penetrates deeper into living tissues and causes less damage. Furthermore, 2-photon signals provide additional molecular-level information that 1- photon signals lack. Many years have gone into developing better fluorescent proteins and biosensors for linear imaging. Unfortunately, directly optimizing probes for the 2-photon nonlinear properties has been beyond the reach of biosensor engineers because of the overwhelming complexity of the current state-of-the-art 2-photon characterization. This is a critical impediment for the BRAIN initiative, because robust 2-photon probes with high brightness and stability are needed. Here we describe a three year program to develop a new process and new instruments using available ultrafast lasers and advanced experimental designs that will empower biosensor engineers involved in the BRAIN project to evolve the next generation of multi photon probes.

描述（由申请人提供）：北极光合作开发更好的 2 光子探测器将结合来自蒙大拿州和加拿大的四位截然不同的研究人员和实验室的专业知识以及互补的兴趣和技能，以解决生物活动成像的基本问题脑。现在人们普遍认为，基因编码的荧光生物传感器非常适合提供对大脑中特定细胞的活动进行成像所需的特异性和多功能性。同样清楚的是，为了对厚层或整个大脑中的细胞进行成像，有必要转向 2 光子（非线性）成像，它可以更深入地渗透到活体组织中并造成更少的损伤。此外，2 光子信号提供了 1 光子信号所缺乏的额外分子水平信息。多年来，人们一直致力于开发更好的荧光蛋白和用于线性成像的生物传感器。不幸的是，由于当前最先进的 2 光子表征极其复杂，直接优化 2 光子非线性特性的探针已经超出了生物传感器工程师的能力范围。这是 BRAIN 计划的一个关键障碍，因为需要具有高亮度和稳定性的强大的 2 光子探针。在这里，我们描述了一个为期三年的计划，利用现有的超快激光器和先进的实验设计来开发新工艺和新仪器，这将使参与 BRAIN 项目的生物传感器工程师能够开发下一代多光子探针。

项目成果

A single-phase flow microfluidic cell sorter for multiparameter screening to assist the directed evolution of Ca2+ sensors.

用于多参数筛选的单相流微流体细胞分选仪，以协助 Ca2 传感器的定向进化。

• 发表时间: 2019
• 影响因子: 6.1
• 作者: Zhao, Yufeng;Zhang, Wei;Zhao, Yongxin;Campbell, Robert E;Harrison, D Jed
• 通讯作者: Harrison, D Jed

High throughput instrument to screen fluorescent proteins under two-photon excitation.

用于在双光子激发下筛选荧光蛋白的高通量仪器。

• 发表时间: 2020-12-01
• 影响因子: 3.4
• 作者: Molina, Rosana S;King, Jonathan;Franklin, Jacob;Clack, Nathan;McRaven, Christopher;Goncharov, Vasily;Flickinger, Daniel;Svoboda, Karel;Drobizhev, Mikhail;Hughes, Thomas E
• 通讯作者: Hughes, Thomas E

Deciphering the molecular mechanism responsible for GCaMP6m's Ca2+-dependent change in fluorescence.

破译 GCaMP6m 的 Ca2 依赖性荧光变化的分子机制。

• 发表时间: 2017
• 影响因子: 3.7
• 作者: Barnett, Lauren M;Hughes, Thomas E;Drobizhev, Mikhail
• 通讯作者: Drobizhev, Mikhail

Characterizing the Two-photon Absorption Properties of Fluorescent Molecules in the 680-1300 nm Spectral Range.

表征 680-1300 nm 光谱范围内荧光分子的双光子吸收特性。

• 发表时间: 2020-01-20
• 影响因子: 0.8
• 作者: Drobizhev, Mikhail;Molina, Rosana S;Hughes, Thomas E
• 通讯作者: Hughes, Thomas E

Multiphoton Bleaching of Red Fluorescent Proteins and the Ways to Reduce It.

红色荧光蛋白的多光子漂白及其减少方法。

• 发表时间: 2022-01-11
• 影响因子: 5.6
• 作者: Drobizhev, Mikhail;Molina, Rosana S;Franklin, Jacob
• 通讯作者: Franklin, Jacob