Massively multiplexed imaging probes for comprehensive single-cell analysis
Name
1119539098-MIT.pdf
Size
42.72 MB
Format
Adobe PDF
Checksum (MD5)
bb433c5456ef94ec10784de324848a7f
Author(s)
Kwok, Sheldon J. J.
Advisor(s)
Seok-Hyun Yun.
Date Issued
2019
Publisher
Massachusetts Institute of Technology
Abstract
Optical microscopy techniques are widely used to study cellular physiology in their native tissue environments. In particular, the use of fluorescent probes to tag different cell populations, subcellular compartments, specific proteins and nucleotide sequences has enabled examination of cellular phenotypes with increasingly sophisticated detail. Recent efforts to combine physiological imaging and single-cell molecular analysis seek complete understanding of cellular identity and function within complex tissues. Specific cells of interest can be selected and isolated from tissues for downstream molecular analyses using techniques such as laser capture micro-dissection, or cell tagging with photo-conversion. However, high-throughput, unbiased molecular profiling of every cell imaged within a tissue remains an elusive challenge. A fundamental obstacle in previous approaches is spectral overlap due to the relatively broad emission of typical fluorescent probes, which limits their capabilities for multiplexed tagging. The first part of this thesis describes methods for studying cellular physiology in mice at single-cell resolution using two-photon fluorescence microscopy. The second part of this thesis describes the development of a novel class of imaging probes, called laser particles, which rely on narrowband laser emission for massively multiplexed cell tagging. This work establishes laser particles as promising tools for comprehensive single-cell analyses.
Description
Thesis: Ph. D. in Medical Engineering and Medical Physics, Harvard-MIT Program in Health Sciences and Technology, 2019
Cataloged from PDF version of thesis.
Includes bibliographical references (pages [165]-191).
Subjects
Harvard--MIT Program in Health Sciences and Technology.
MIT Department
Harvard--MIT Program in Health Sciences and Technology
Harvard University--MIT Division of Health Sciences and Technology
Terms of Use
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