Fluorescence is the current standard method of choice for intravital microscopy and cytometry. However, the broad emission spectrum of fluorescent probes?dyes, fluorescent probes, and quantum dots?limits the number of cells that can be tracked simultaneously without ambiguity. DNA barcodes can label cells but cannot be visualized in vivo, as they require in vitro genetic reading. The optical principle of stimulated emission and cavity resonance used in a laser can generate extremely narrow spectral line-widths over a broad spectral range. This project will miniaturize lasers to the sizes of mitochondria or viruses and develop instrumentations to utilize the laser particles as novel probes for massively parallel imaging and assays. By tracing individual cells over time in mice, the proliferation, migrations and cell-cell and cell-tissue interactions can be studied in vivo. The cells can be further analyzed by flow cytometry and sorted for gene profiling and single-cell RNA sequencing, providing comprehensive information from molecular, cellular, tissue, and systems levels over millions to billions of cells in a single animal experiment. The first specific aim is to create a new paradigm for imaging-compatible cellular labeling using injectable, biocompatible micro- and submicron-cavity lasers.
The second aim i s to develop Laser Particle Stimulated Emission (LASE) Microscopy for conducting labeled microscopy in vivo at depths of up to 3 mm. The third specific aim is to demonstrate massively multiplexed, high-throughput cell tracking and analysis. The breakthrough capabilities will be used to dissect tumor heterogeneity in progression, metastasis, and response to therapy at unprecedented single-cell resolution.

Public Health Relevance

This proposal is relevant to public health because it will develop novel imaging and high- throughput analysis method to study the molecular and cellular mechanisms of diseases and their therapeutic responses. Therefore, the proposed research is relevant to the NIH's mission of fostering innovative research strategies to increase the nation's ability to improve the treatment of disease.

Agency
National Institute of Health (NIH)
Institute
National Institute of Biomedical Imaging and Bioengineering (NIBIB)
Type
NIH Director’s Pioneer Award (NDPA) (DP1)
Project #
5DP1EB024242-04
Application #
9767768
Study Section
Special Emphasis Panel (ZRG1)
Program Officer
King, Randy Lee
Project Start
2016-09-30
Project End
2021-07-31
Budget Start
2019-08-01
Budget End
2020-07-31
Support Year
4
Fiscal Year
2019
Total Cost
Indirect Cost
Name
Massachusetts General Hospital
Department
Type
DUNS #
073130411
City
Boston
State
MA
Country
United States
Zip Code
02114
Humar, Matjaž; Upadhya, Avinash; Yun, Seok Hyun (2017) Spectral reading of optical resonance-encoded cells in microfluidics. Lab Chip 17:2777-2784
Humar, Matjaž; Dobravec, Anja; Zhao, Xiangwei et al. (2017) Biomaterial microlasers implantable in the cornea, skin, and blood. Optica 4:1080-1085
Yun, Seok Hyun; Kwok, Sheldon J J (2017) Light in diagnosis, therapy and surgery. Nat Biomed Eng 1:
Humar, Matjaž; Yun, Seok Hyun (2017) Whispering-gallery-mode emission from biological luminescent protein microcavity assemblies. Optica 4:222-228
Cho, Sangyeon; Humar, Matjaž; Martino, Nicola et al. (2016) Laser Particle Stimulated Emission Microscopy. Phys Rev Lett 117:193902