Laser particle activated cell sorting in microfluidics

Dannenberg, Paul H.; Kang, Jisoo; Martino, Nicola; Kashiparekh, Anokhi; Forward, Sarah; Wu, Jiamin; Liapis, Andreas C.; Wang, Jie; Yun, Seok Hyun

doi:10.1039/d2lc00235c

Cited by 10 publications

(9 citation statements)

References 28 publications

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“…In the future, functionalized microlasers could also be bound to specific cells by targeting surface markers, for example, EpCAM present in carcinomas [51] and specific T cell receptors, [52] where they could help to evaluate cellular heterogeneity in the context of cellular development and for evaluation of drug therapy at the single cell level. [19,53] Beyond high throughput screening, functionalized microlasers may be used to specifically label live cells that present specific antigens with the microlasers serving as optical barcodes. This would be useful to follow the migration of certain cell populations, e.g.…”

Section: Conclusion and Discussionmentioning

confidence: 99%

Section: Conclusion and Discussionmentioning

confidence: 99%

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Local Sensing of Absolute Refractive Index During Protein‐Binding using Microlasers with Spectral Encoding

Caixeiro

Kunstmann‐Olsen

Schubert

et al. 2023

Advanced Optical Materials

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Multiplexed, specific, and sensitive detection of antigens is critical for the rapid and accurate diagnosis of disease and the informed development of personalized treatment plans. Here, it is shown that polymer microsphere lasers can be used as photonic sensors to monitor and quantify direct surface binding of biomolecules via changes in the refractive index. The unique spectral signature of each individual laser can be used to find their size and effective refractive index which adds a new encoding dimension when compared to conventional fluorescent beads. Antibody-functionalized microlasers selectively detect protein binding, as demonstrated for Immunoglobulin G and C-reactive protein, and have the ability to resolve different stages of the multilayer surface modification. Moreover, by continuously monitoring single lasers, the possibility of real-time monitoring of binding dynamics between antigens in solution phase and the immobilized antibodies is demonstrated. For multiplexed detection, the microlasers are employed in a flow cytometer configuration, with fast spectral detection and identification of microlasers with and without antigen binding. It is envisioned that by combining microlasers with well-established surface modification chemistries and flow geometries, the multiplexing ability of microbead immunoassays can be strongly increased while also opening avenues for single-cell profiling within heterogeneous cell populations.

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Section: Conclusion and Discussionmentioning

confidence: 99%

Section: Conclusion and Discussionmentioning

confidence: 99%

Local Sensing of Absolute Refractive Index During Protein‐Binding using Microlasers with Spectral Encoding

Caixeiro

Kunstmann‐Olsen

Schubert

et al. 2023

Advanced Optical Materials

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“…For applications not requiring spatial information of the sample, a noteworthy alternative to the imaging-based approaches is spectral acquisition in a micro uidic con guration 45 . Integrating a line-scan camera in such a system has enabled spectral acquisition at rates of 25-30 kHz 46 .…”

Section: Development Of the Protocol And Comparison With Other Methodsmentioning

confidence: 99%

Hyperspectral Confocal Imaging for High-Throughput Readout and Analysis of Bio-Integrated Laser Particles

Titze

Caixeiro

Dinh

et al. 2023

Preprint

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Integrating laser particles into live cells, tissue cultures, and small animals is an emerging and rapidly evolving technique that offers non-invasive interrogation and labelling with unprecedented information density. The bright and distinct spectra of laser particles make this approach particularly attractive for high-throughput applications requiring single-cell specificity, such as multiplexed cell tracking and intracellular biosensing. To be of practical relevance, 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 protocol consisting of two separate procedures. The first part of our protocol instructs users on how to efficiently integrate different types of laser particles into living cells. The second part 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 design choices and recommend specific workflows for processing the resulting large data sets along with an open-source Python library of functions covering the entire analysis pipeline. The results one can obtain using this protocol are illustrated with three representative experiments: Rapid, large-volume mapping of absolute refractive index using polystyrene microbead laser particles, intracellular sensing to monitor cardiac contractility with polystyrene microbead laser particles, and long-term cell tracking using semiconductor nanodisk laser particles. Our sample preparation and imaging procedures can be completed within two days. Setting up the hyperspectral confocal microscope for laser particle characterization will take users with little prior experience in optical and software engineering <2 weeks to complete.

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“…Cell sorting in microfluidics based on the intracellular microcavity spectra at a kilohertz rate was also demonstrated. 112 In Tissues and in Vivo. Long-term tracking of a large number of cells in vivo is, in general, extremely challenging.…”

Section: Acsmentioning

confidence: 99%

“…A larger number of parameters without overlap was also demonstrated, by first measuring one set of parameters, then photobleaching the probes, restaining, and measuring the second set of parameters. Cell sorting in microfluidics based on the intracellular microcavity spectra at a kilohertz rate was also demonstrated …”

Section: Applicationsmentioning

confidence: 99%

Microcavity- and Microlaser-Based Optical Barcoding: A Review of Encoding Techniques and Applications

2023

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Optical microbarcodes have recently received a great deal of interest because of their suitability for a wide range of applications, such as multiplexed assays, cell tagging and tracking, anticounterfeiting, and product labeling. Spectral barcodes are especially promising because they are robust and have a simple readout. In addition, microcavity- and microlaser-based barcodes have very narrow spectra and therefore have the potential to generate millions of unique barcodes. This review begins with a discussion of the different types of barcodes and then focuses specifically on microcavity-based barcodes. While almost any kind of optical microcavity can be used for barcoding, currently whispering-gallery microcavities (in the form of spheres and disks), nanowire lasers, Fabry–Pérot lasers, random lasers, and distributed feedback lasers are the most frequently employed for this purpose. In microcavity-based barcodes, the information is encoded in various ways in the properties of the emitted light, most frequently in the spectrum. The barcode is dependent on the properties of the microcavity, such as the size, shape, and the gain materials. Various applications of these barcodes, including cell tracking, anticounterfeiting, and product labeling are described. Finally, the future prospects for microcavity- and microlaser-based barcodes are discussed.

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Laser particle activated cell sorting in microfluidics

Abstract: A microfluidic device is developed that uses the narrowband optical radiation of laser particle barcodes to sort tagged cells.

Cited by 10 publications

References 28 publications

Local Sensing of Absolute Refractive Index During Protein‐Binding using Microlasers with Spectral Encoding

Local Sensing of Absolute Refractive Index During Protein‐Binding using Microlasers with Spectral Encoding

Hyperspectral Confocal Imaging for High-Throughput Readout and Analysis of Bio-Integrated Laser Particles

Microcavity- and Microlaser-Based Optical Barcoding: A Review of Encoding Techniques and Applications

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