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Article: Microfluidic Imaging Flow Cytometry by Asymmetric-detection Time-stretch Optical Microscopy (ATOM)

TitleMicrofluidic Imaging Flow Cytometry by Asymmetric-detection Time-stretch Optical Microscopy (ATOM)
Authors
KeywordsEngineering
Issue 124
Microscopy
Imaging flow cytometry
Microfluidic
Time-stretch imaging
High-throughput screening
Single-cell analysis
Issue Date2017
PublisherJournal of Visualized Experiments. The Journal's web site is located at http://www.jove.com
Citation
Journal of Visualized Experiments, 2017, n. 124, article no. e55840 How to Cite?
AbstractScaling the number of measurable parameters, which allows for multidimensional data analysis and thus higher-confidence statistical results, has been the main trend in the advanced development of flow cytometry. Notably, adding high-resolution imaging capabilities allows for the complex morphological analysis of cellular/sub-cellular structures. This is not possible with standard flow cytometers. However, it is valuable for advancing our knowledge of cellular functions and can benefit life science research, clinical diagnostics, and environmental monitoring. Incorporating imaging capabilities into flow cytometry compromises the assay throughput, primarily due to the limitations on speed and sensitivity in the camera technologies. To overcome this speed or throughput challenge facing imaging flow cytometry while preserving the image quality, asymmetric-detection time-stretch optical microscopy (ATOM) has been demonstrated to enable high-contrast, single-cell imaging with sub-cellular resolution, at an imaging throughput as high as 100,000 cells/s. Based on the imaging concept of conventional time-stretch imaging, which relies on all-optical image encoding and retrieval through the use of ultrafast broadband laser pulses, ATOM further advances imaging performance by enhancing the image contrast of unlabeled/unstained cells. This is achieved by accessing the phase-gradient information of the cells, which is spectrally encoded into single-shot broadband pulses. Hence, ATOM is particularly advantageous in high-throughput measurements of single-cell morphology and texture – information indicative of cell types, states, and even functions. Ultimately, this could become a powerful imaging flow cytometry platform for the biophysical phenotyping of cells, complementing the current state-of-the-art biochemical-marker-based cellular assay. This work describes a protocol to establish the key modules of an ATOM system (from optical frontend to data processing and visualization backend), as well as the workflow of imaging flow cytometry based on ATOM, using human cells and micro-algae as the examples.
Persistent Identifierhttp://hdl.handle.net/10722/244985
ISSN
2019 Impact Factor: 1.163
2015 SCImago Journal Rankings: 0.451
PubMed Central ID
ISI Accession Number ID

 

DC FieldValueLanguage
dc.contributor.authorTang, AHL-
dc.contributor.authorLai, QTK-
dc.contributor.authorChung, BMF-
dc.contributor.authorLee, KCM-
dc.contributor.authorMok, ATY-
dc.contributor.authorYip, GK-
dc.contributor.authorShum, AHC-
dc.contributor.authorWong, KKY-
dc.contributor.authorTsia, KK-
dc.date.accessioned2017-09-18T02:02:36Z-
dc.date.available2017-09-18T02:02:36Z-
dc.date.issued2017-
dc.identifier.citationJournal of Visualized Experiments, 2017, n. 124, article no. e55840-
dc.identifier.issn1940-087X-
dc.identifier.urihttp://hdl.handle.net/10722/244985-
dc.description.abstractScaling the number of measurable parameters, which allows for multidimensional data analysis and thus higher-confidence statistical results, has been the main trend in the advanced development of flow cytometry. Notably, adding high-resolution imaging capabilities allows for the complex morphological analysis of cellular/sub-cellular structures. This is not possible with standard flow cytometers. However, it is valuable for advancing our knowledge of cellular functions and can benefit life science research, clinical diagnostics, and environmental monitoring. Incorporating imaging capabilities into flow cytometry compromises the assay throughput, primarily due to the limitations on speed and sensitivity in the camera technologies. To overcome this speed or throughput challenge facing imaging flow cytometry while preserving the image quality, asymmetric-detection time-stretch optical microscopy (ATOM) has been demonstrated to enable high-contrast, single-cell imaging with sub-cellular resolution, at an imaging throughput as high as 100,000 cells/s. Based on the imaging concept of conventional time-stretch imaging, which relies on all-optical image encoding and retrieval through the use of ultrafast broadband laser pulses, ATOM further advances imaging performance by enhancing the image contrast of unlabeled/unstained cells. This is achieved by accessing the phase-gradient information of the cells, which is spectrally encoded into single-shot broadband pulses. Hence, ATOM is particularly advantageous in high-throughput measurements of single-cell morphology and texture – information indicative of cell types, states, and even functions. Ultimately, this could become a powerful imaging flow cytometry platform for the biophysical phenotyping of cells, complementing the current state-of-the-art biochemical-marker-based cellular assay. This work describes a protocol to establish the key modules of an ATOM system (from optical frontend to data processing and visualization backend), as well as the workflow of imaging flow cytometry based on ATOM, using human cells and micro-algae as the examples.-
dc.languageeng-
dc.publisherJournal of Visualized Experiments. The Journal's web site is located at http://www.jove.com-
dc.relation.ispartofJournal of Visualized Experiments-
dc.rightsCopyright © 2017 Journal of Visualized Experiments.-
dc.subjectEngineering-
dc.subjectIssue 124-
dc.subjectMicroscopy-
dc.subjectImaging flow cytometry-
dc.subjectMicrofluidic-
dc.subjectTime-stretch imaging-
dc.subjectHigh-throughput screening-
dc.subjectSingle-cell analysis-
dc.titleMicrofluidic Imaging Flow Cytometry by Asymmetric-detection Time-stretch Optical Microscopy (ATOM)-
dc.typeArticle-
dc.identifier.emailShum, AHC: ashum@hku.hk-
dc.identifier.emailWong, KKY: kywong@eee.hku.hk-
dc.identifier.emailTsia, KK: tsia@hku.hk-
dc.identifier.authorityShum, AHC=rp01439-
dc.identifier.authorityWong, KKY=rp00189-
dc.identifier.authorityTsia, KK=rp01389-
dc.description.naturepublished_or_final_version-
dc.identifier.doi10.3791/55840-
dc.identifier.pmid28715367-
dc.identifier.pmcidPMC5608519-
dc.identifier.scopuseid_2-s2.0-85025142125-
dc.identifier.hkuros278673-
dc.identifier.issue124-
dc.identifier.spagearticle no. e55840-
dc.identifier.epagearticle no. e55840-
dc.identifier.isiWOS:000407448100061-
dc.publisher.placeUnited States-

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