Hyperspectral confocal imaging for high-throughput readout and analysis of bio-integrated microlasers
Date
18/01/2024Author
Funder
Grant ID
640012
659213
DH160102
RGF/R1/180070
RGF/EA/180051
EP/P030017/1
Metadata
Show full item recordAbstract
Integrating micro- and nanolasers into live cells, tissue cultures and small animals is an emerging and rapidly evolving technique that offers noninvasive interrogation and labeling with unprecedented information density. The bright and distinct spectra of such lasers make this approach particularly attractive for high-throughput applications requiring single-cell specificity, such as multiplexed cell tracking and intracellular biosensing. The implementation of these applications requires high-resolution, high-speed spectral readout and advanced analysis routines, which leads to unique technical challenges. Here, we present a modular approach consisting of two separate procedures. The first procedure instructs users on how to efficiently integrate different types of lasers into living cells, and the second procedure presents a workflow for obtaining intracellular lasing spectra with high spectral resolution and up to 125-kHz readout rate and starts from the construction of a custom hyperspectral confocal microscope. We provide guidance on running hyperspectral imaging routines for various experimental designs and recommend specific workflows for processing the resulting large data sets along with an open-source Python library of functions covering the analysis pipeline. We illustrate three applications including the rapid, large-volume mapping of absolute refractive index by using polystyrene microbead lasers, the intracellular sensing of cardiac contractility with polystyrene microbead lasers and long-term cell tracking by using semiconductor nanodisk lasers. Our sample preparation and imaging procedures require 2 days, and setting up the hyperspectral confocal microscope for microlaser characterization requires
Citation
Titze , V M , Caixeiro , S , Dinh , V S , König , M , Rübsam , M , Pathak , N , Schumacher , A-L , Germer , M , Kukat , C , Niessen , C M , Schubert , M & Gather , M C 2024 , ' Hyperspectral confocal imaging for high-throughput readout and analysis of bio-integrated microlasers ' , Nature Protocols . https://doi.org/10.1038/s41596-023-00924-6
Publication
Nature Protocols
Status
Peer reviewed
ISSN
1754-2189Type
Journal article
Description
Funding: This work received financial support from the Leverhulme Trust (RPG-2017-231), the European Union’s Horizon 2020 Framework Programme (FP/2014-2020)/ERC grant agreement no. 640012 (ABLASE), EPSRC (EP/P030017/1), the Humboldt Foundation (Alexander von Humboldt professorship) and the RS Macdonald Charitable Trust (St Andrews Seedcorn Fund for Neurological Research). M.S. acknowledges funding from the European Commission (Marie Skłodowska-Curie Individual Fellowship, 659213) and the Royal Society (Dorothy Hodgkin Fellowship, DH160102; Research Grant, RGF\R1\180070; Enhancement Award, RGF\EA\180051).Collections
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