ACS Publications. Most Trusted. Most Cited. Most Read
A Heating-Superfusion Platform Technology for the Investigation of Protein Function in Single Cells

OR SEARCH CITATIONS

My Activity
Publications

Figure 1Loading Img
Download Hi-Res ImageDownload to MS-PowerPointCite This:Anal. Chem. 2015, 87, 1, 381-387
    Technical Note

    A Heating-Superfusion Platform Technology for the Investigation of Protein Function in Single Cells
    Click to copy article linkArticle link copied!

    View Author Information
    † Department of Chemical and Biological Engineering, Chalmers University of Technology Kemivägen 10, SE-412 96 Gothenburg, Sweden
    ‡ Department of Physiology and Pharmacology, Karolinska Institutet, SE-17177 Stockholm, Sweden
    § Department of Chemistry and Chemical Biology, Harvard University, Cambridge, Massachusetts 02138, United States
    *Tel.: +46 31 772 61 23. E-mail: [email protected]
    Other Access OptionsSupporting Information (1)

    Analytical Chemistry

    Cite this: Anal. Chem. 2015, 87, 1, 381–387
    Click to copy citationCitation copied!
    https://doi.org/10.1021/ac5031418
    Published December 2, 2014
    Copyright © 2014 American Chemical Society
    Request reuse permissions

    Abstract

    Click to copy section linkSection link copied!
    Abstract Image

    Here, we report on a novel approach for the study of single-cell intracellular enzyme activity at various temperatures, utilizing a localized laser heating probe in combination with a freely positionable microfluidic perfusion device. Through directed exposure of individual cells to the pore-forming agent α-hemolysin, we have controlled the membrane permeability, enabling targeted delivery of the substrate. Mildly permeabilized cells were exposed to fluorogenic substrates to monitor the activity of intracellular enzymes, while adjusting the local temperature surrounding the target cells, using an infrared laser heating system. We generated quantitative estimates for the intracellular alkaline phosphatase activity at five different temperatures in different cell lines, constructing temperature-response curves of enzymatic activity at the single-cell level. Enzymatic activity was determined rapidly after cell permeation, generating five-point temperature-response curves within just 200 s.

    Copyright © 2014 American Chemical Society

    Subjects

    what are subjects

    Read this article

    To access this article, please review the available access options below.

    Get instant access

    Purchase Access

    Read this article for 48 hours. Check out below using your ACS ID or as a guest.

    Recommended

    Access through Your Institution

    You may have access to this article through your institution.

    Your institution does not have access to this content. Add or change your institution or let them know you’d like them to include access.

    Supporting Information

    Click to copy section linkSection link copied!

    This material is available free of charge via the Internet at http://pubs.acs.org.

    Terms & Conditions

    Most electronic Supporting Information files are available without a subscription to ACS Web Editions. Such files may be downloaded by article for research use (if there is a public use license linked to the relevant article, that license may permit other uses). Permission may be obtained from ACS for other uses through requests via the RightsLink permission system: http://pubs.acs.org/page/copyright/permissions.html.

    Cited By

    Click to copy section linkSection link copied!

    This article is cited by 9 publications.

    1. Ioannis Zachos, Robert Genth, Samuel Sutiono, Matthias Marczynski, Oliver Lieleg, Volker Sieber. Hot Flows: Evolving an Archaeal Glucose Dehydrogenase for Ultrastable Carba-NADP+ Using Microfluidics at Elevated Temperatures. ACS Catalysis 2022, 12 (3) , 1841-1846. https://doi.org/10.1021/acscatal.1c04320
    2. Sanjin Hosic, Shashi K. Murthy, and Abigail N. Koppes . Microfluidic Sample Preparation for Single Cell Analysis. Analytical Chemistry 2016, 88 (1) , 354-380. https://doi.org/10.1021/acs.analchem.5b04077
    3. Michael G. Roper . Cellular Analysis Using Microfluidics. Analytical Chemistry 2016, 88 (1) , 381-394. https://doi.org/10.1021/acs.analchem.5b04532
    4. Zengnan Wu, Ling Lin. Nanofluidics for single-cell analysis. Chinese Chemical Letters 2022, 33 (4) , 1752-1756. https://doi.org/10.1016/j.cclet.2021.08.100
    5. Aldo Jesorka, Inga Põldsalu, Irep Gözen. Microfluidic technology for investigation of protein function in single adherent cells. 2019, 145-172. https://doi.org/10.1016/bs.mie.2019.07.038
    6. Mashooq Khan, Sifeng Mao, Weiwei Li, Jin‐Ming Lin. Microfluidic Devices in the Fast‐Growing Domain of Single‐Cell Analysis. Chemistry – A European Journal 2018, 24 (58) , 15398-15420. https://doi.org/10.1002/chem.201800305
    7. Ling Lin, Qinghua Chen, Jiashu Sun. Micro/nanofluidics-enabled single-cell biochemical analysis. TrAC Trends in Analytical Chemistry 2018, 99 , 66-74. https://doi.org/10.1016/j.trac.2017.11.017
    8. Irep Gözen, Gavin Jeffries, Tatsiana Lobovkina, Emanuele Celauro, Mehrnaz Shaali, Baharan Ali Doosti, Aldo Jesorka. Single-Cell Analysis with the BioPen. 2018, 187-219. https://doi.org/10.1002/9783527696789.ch10
    9. Aldo Jesorka, Irep Gözen. The Multifunctional Pipette. 2018, 155-185. https://doi.org/10.1002/9783527696789.ch9
    Download PDF
    Go to Analytical Chemistry
    Get e-Alerts
    Get e-Alerts

    Analytical Chemistry

    Cite this: Anal. Chem. 2015, 87, 1, 381–387
    Click to copy citationCitation copied!
    https://doi.org/10.1021/ac5031418
    Published December 2, 2014
    Copyright © 2014 American Chemical Society
    Request reuse permissions

    Article Views

    517

    Altmetric

    -

    Citations

    9
    Learn about these metrics

    Article Views are the COUNTER-compliant sum of full text article downloads since November 2008 (both PDF and HTML) across all institutions and individuals. These metrics are regularly updated to reflect usage leading up to the last few days.

    Citations are the number of other articles citing this article, calculated by Crossref and updated daily. Find more information about Crossref citation counts.

    The Altmetric Attention Score is a quantitative measure of the attention that a research article has received online. Clicking on the donut icon will load a page at altmetric.com with additional details about the score and the social media presence for the given article. Find more information on the Altmetric Attention Score and how the score is calculated.