Spatially organized fluorescent reporters for recording complex biological dynamics in cell population

Biological signals, such as the dynamic concentrations of ions, levels of signaling molecules, and activities of protein kinases, interact in complex ways within cells, and can exhibit great cell-to-cell heterogeneity as a function of cell history and state. There is increasing desire to use multiple fluorescent reporters to simultaneously measure multiple biological signals in single cells across cell populations, such as those in the brain. However, due to the diffraction limit of optical imaging, the biological signals recorded from neurons in densely-labeled neural populations in vivo are often mixed with signals from closely passing axons and dendrites from other neurons, resulting in erroneous signaling events and artifactual correlations of measured neural activity. Also, it is not yet possible to simultaneously record any given set of biological signals in single cells, because there are limited sets of corresponding spectrally-orthogonal fluorescent reporters available to date. Even if the fluorescent reporters for all biological signals in all possible colors are developed in the future, the number of biological signals that can be simultaneously recorded are still limited by the number of available optical channels. In this thesis, I address these problems by developing two new technologies, soma-targeted fluorescent calcium indicators and spatially multiplexed imaging.

Name thesis:degree_name

Doctoral

Department dc:contributor.department

Massachusetts Institute of Technology. Department of Electrical Engineering and Computer Science

Grantor dc:publisher

Massachusetts Institute of Technology

Year dc:date.issued

2020

Author dc:creator

• Linghu, Changyang

Advisor dc:contributor.advisor

• Edward S. Boyden.

• Electrical Engineering and Computer Science.

Statement dc:rights

• MIT theses may be protected by copyright. Please reuse MIT thesis content according to the MIT Libraries Permissions Policy, which is available through the URL provided.

Licence dc:rights.uri

• http://dspace.mit.edu/handle/1721.1/7582

Language dc:language.iso

eng

Handle dc:identifier.uri

https://hdl.handle.net/1721.1/128587

OAI identifier oai:identifier

oai:dspace.mit.edu:1721.1/128587