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Article: Microfluidic Imaging Flow Cytometry by Asymmetric-detection Time-stretch Optical Microscopy (ATOM)
Title | Microfluidic Imaging Flow Cytometry by Asymmetric-detection Time-stretch Optical Microscopy (ATOM) |
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Authors | |
Keywords | Engineering Issue 124 Microscopy Imaging flow cytometry Microfluidic Time-stretch imaging High-throughput screening Single-cell analysis |
Issue Date | 2017 |
Publisher | Journal 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? |
Abstract | Scaling 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 Identifier | http://hdl.handle.net/10722/244985 |
ISSN | 2023 Impact Factor: 1.2 2023 SCImago Journal Rankings: 0.449 |
PubMed Central ID | |
ISI Accession Number ID |
DC Field | Value | Language |
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dc.contributor.author | Tang, AHL | - |
dc.contributor.author | Lai, QTK | - |
dc.contributor.author | Chung, BMF | - |
dc.contributor.author | Lee, KCM | - |
dc.contributor.author | Mok, ATY | - |
dc.contributor.author | Yip, GK | - |
dc.contributor.author | Shum, AHC | - |
dc.contributor.author | Wong, KKY | - |
dc.contributor.author | Tsia, KK | - |
dc.date.accessioned | 2017-09-18T02:02:36Z | - |
dc.date.available | 2017-09-18T02:02:36Z | - |
dc.date.issued | 2017 | - |
dc.identifier.citation | Journal of Visualized Experiments, 2017, n. 124, article no. e55840 | - |
dc.identifier.issn | 1940-087X | - |
dc.identifier.uri | http://hdl.handle.net/10722/244985 | - |
dc.description.abstract | Scaling 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.language | eng | - |
dc.publisher | Journal of Visualized Experiments. The Journal's web site is located at http://www.jove.com | - |
dc.relation.ispartof | Journal of Visualized Experiments | - |
dc.rights | Copyright © 2017 Journal of Visualized Experiments. | - |
dc.subject | Engineering | - |
dc.subject | Issue 124 | - |
dc.subject | Microscopy | - |
dc.subject | Imaging flow cytometry | - |
dc.subject | Microfluidic | - |
dc.subject | Time-stretch imaging | - |
dc.subject | High-throughput screening | - |
dc.subject | Single-cell analysis | - |
dc.title | Microfluidic Imaging Flow Cytometry by Asymmetric-detection Time-stretch Optical Microscopy (ATOM) | - |
dc.type | Article | - |
dc.identifier.email | Shum, AHC: ashum@hku.hk | - |
dc.identifier.email | Wong, KKY: kywong@eee.hku.hk | - |
dc.identifier.email | Tsia, KK: tsia@hku.hk | - |
dc.identifier.authority | Shum, AHC=rp01439 | - |
dc.identifier.authority | Wong, KKY=rp00189 | - |
dc.identifier.authority | Tsia, KK=rp01389 | - |
dc.description.nature | published_or_final_version | - |
dc.identifier.doi | 10.3791/55840 | - |
dc.identifier.pmid | 28715367 | - |
dc.identifier.pmcid | PMC5608519 | - |
dc.identifier.scopus | eid_2-s2.0-85025142125 | - |
dc.identifier.hkuros | 278673 | - |
dc.identifier.issue | 124 | - |
dc.identifier.spage | article no. e55840 | - |
dc.identifier.epage | article no. e55840 | - |
dc.identifier.isi | WOS:000407448100061 | - |
dc.publisher.place | United States | - |
dc.identifier.issnl | 1940-087X | - |