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1.
Nat Commun ; 11(1): 3452, 2020 07 10.
Artículo en Inglés | MEDLINE | ID: mdl-32651381

RESUMEN

The advent of image-activated cell sorting and imaging-based cell picking has advanced our knowledge and exploitation of biological systems in the last decade. Unfortunately, they generally rely on fluorescent labeling for cellular phenotyping, an indirect measure of the molecular landscape in the cell, which has critical limitations. Here we demonstrate Raman image-activated cell sorting by directly probing chemically specific intracellular molecular vibrations via ultrafast multicolor stimulated Raman scattering (SRS) microscopy for cellular phenotyping. Specifically, the technology enables real-time SRS-image-based sorting of single live cells with a throughput of up to ~100 events per second without the need for fluorescent labeling. To show the broad utility of the technology, we show its applicability to diverse cell types and sizes. The technology is highly versatile and holds promise for numerous applications that are previously difficult or undesirable with fluorescence-based technologies.


Asunto(s)
Separación Celular/métodos , Espectrometría Raman/métodos , Animales , Humanos
2.
Proc Natl Acad Sci U S A ; 116(32): 15842-15848, 2019 08 06.
Artículo en Inglés | MEDLINE | ID: mdl-31324741

RESUMEN

Combining the strength of flow cytometry with fluorescence imaging and digital image analysis, imaging flow cytometry is a powerful tool in diverse fields including cancer biology, immunology, drug discovery, microbiology, and metabolic engineering. It enables measurements and statistical analyses of chemical, structural, and morphological phenotypes of numerous living cells to provide systematic insights into biological processes. However, its utility is constrained by its requirement of fluorescent labeling for phenotyping. Here we present label-free chemical imaging flow cytometry to overcome the issue. It builds on a pulse pair-resolved wavelength-switchable Stokes laser for the fastest-to-date multicolor stimulated Raman scattering (SRS) microscopy of fast-flowing cells on a 3D acoustic focusing microfluidic chip, enabling an unprecedented throughput of up to ∼140 cells/s. To show its broad utility, we use the SRS imaging flow cytometry with the aid of deep learning to study the metabolic heterogeneity of microalgal cells and perform marker-free cancer detection in blood.


Asunto(s)
Citometría de Flujo/métodos , Imagenología Tridimensional , Espectrometría Raman/métodos , Línea Celular Tumoral , Humanos , Microalgas/citología , Microalgas/metabolismo , Coloración y Etiquetado
3.
Sci Adv ; 5(1): eaau0241, 2019 01.
Artículo en Inglés | MEDLINE | ID: mdl-30746443

RESUMEN

Flow cytometry is an indispensable tool in biology for counting and analyzing single cells in large heterogeneous populations. However, it predominantly relies on fluorescent labeling to differentiate cells and, hence, comes with several fundamental drawbacks. Here, we present a high-throughput Raman flow cytometer on a microfluidic chip that chemically probes single live cells in a label-free manner. It is based on a rapid-scan Fourier-transform coherent anti-Stokes Raman scattering spectrometer as an optical interrogator, enabling us to obtain the broadband molecular vibrational spectrum of every single cell in the fingerprint region (400 to 1600 cm-1) with a record-high throughput of ~2000 events/s. As a practical application of the method not feasible with conventional flow cytometry, we demonstrate high-throughput label-free single-cell analysis of the astaxanthin productivity and photosynthetic dynamics of Haematococcus lacustris.


Asunto(s)
Citometría de Flujo/métodos , Espectrometría Raman/métodos , Dióxido de Carbono/metabolismo , Isótopos de Carbono/metabolismo , Chlorophyceae/metabolismo , Citometría de Flujo/instrumentación , Análisis de Fourier , Ensayos Analíticos de Alto Rendimiento/instrumentación , Ensayos Analíticos de Alto Rendimiento/métodos , Dispositivos Laboratorio en un Chip , Fotosíntesis , Reproducibilidad de los Resultados , Análisis de la Célula Individual/instrumentación , Análisis de la Célula Individual/métodos , Espectrometría Raman/instrumentación , Vibración , Xantófilas/metabolismo
4.
Cell ; 175(1): 266-276.e13, 2018 09 20.
Artículo en Inglés | MEDLINE | ID: mdl-30166209

RESUMEN

A fundamental challenge of biology is to understand the vast heterogeneity of cells, particularly how cellular composition, structure, and morphology are linked to cellular physiology. Unfortunately, conventional technologies are limited in uncovering these relations. We present a machine-intelligence technology based on a radically different architecture that realizes real-time image-based intelligent cell sorting at an unprecedented rate. This technology, which we refer to as intelligent image-activated cell sorting, integrates high-throughput cell microscopy, focusing, and sorting on a hybrid software-hardware data-management infrastructure, enabling real-time automated operation for data acquisition, data processing, decision-making, and actuation. We use it to demonstrate real-time sorting of microalgal and blood cells based on intracellular protein localization and cell-cell interaction from large heterogeneous populations for studying photosynthesis and atherothrombosis, respectively. The technology is highly versatile and expected to enable machine-based scientific discovery in biological, pharmaceutical, and medical sciences.


Asunto(s)
Citometría de Flujo/métodos , Ensayos Analíticos de Alto Rendimiento/métodos , Procesamiento de Imagen Asistido por Computador/métodos , Animales , Aprendizaje Profundo , Humanos
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