Labeling and tracking of immune cells in ex vivo human skin
Human skin harbors various immune cells that are crucial for the control of injury and infection. However, the current understanding of immune cell function within viable human skin tissue is limited. We developed an ex vivo imaging approach in which fresh skin biopsies are mounted and then labeled...
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| Published in: | Nature protocols Vol. 16; no. 2; pp. 791 - 811 |
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| Main Authors: | , , , , , , , , |
| Format: | Journal Article |
| Language: | English |
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Nature Publishing Group UK
01.02.2021
Nature Publishing Group |
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| ISSN: | 1754-2189, 1750-2799, 1750-2799 |
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| Abstract | Human skin harbors various immune cells that are crucial for the control of injury and infection. However, the current understanding of immune cell function within viable human skin tissue is limited. We developed an ex vivo imaging approach in which fresh skin biopsies are mounted and then labeled with nanobodies or antibodies against cell surface markers on tissue-resident memory CD8
+
T cells, other immune cells of interest, or extracellular tissue components. Subsequent longitudinal imaging allows one to describe the dynamic behavior of human skin-resident cells in situ. In addition, this strategy can be used to study immune cell function in murine skin. The ability to follow the spatiotemporal behavior of CD8
+
T cells and other immune cells in skin, including their response to immune stimuli, provides a platform to investigate physiological immune cell behavior and immune cell behavior in skin diseases. The mounting, staining and imaging of skin samples requires ~1.5 d, and subsequent tracking analysis requires a minimum of 1 d. The optional production of fluorescently labeled nanobodies takes ~5 d.
This protocol describes how to follow the behavior of immune cells by imaging ex vivo cultured human or mouse skin biopsies following labeling with antibody or nanobody reagents against specific cell surface markers. |
|---|---|
| AbstractList | Human skin harbors various immune cells that are crucial for the control of injury and infection. However, the current understanding of immune cell function within viable human skin tissue is limited. We developed an ex vivo imaging approach in which fresh skin biopsies are mounted and then labeled with nanobodies or antibodies against cell surface markers on tissue-resident memory CD8.sup.+ T cells, other immune cells of interest, or extracellular tissue components. Subsequent longitudinal imaging allows one to describe the dynamic behavior of human skin-resident cells in situ. In addition, this strategy can be used to study immune cell function in murine skin. The ability to follow the spatiotemporal behavior of CD8.sup.+ T cells and other immune cells in skin, including their response to immune stimuli, provides a platform to investigate physiological immune cell behavior and immune cell behavior in skin diseases. The mounting, staining and imaging of skin samples requires ~1.5 d, and subsequent tracking analysis requires a minimum of 1 d. The optional production of fluorescently labeled nanobodies takes ~5 d. This protocol describes how to follow the behavior of immune cells by imaging ex vivo cultured human or mouse skin biopsies following labeling with antibody or nanobody reagents against specific cell surface markers. Human skin harbors various immune cells that are crucial for the control of injury and infection. However, the current understanding of immune cell function within viable human skin tissue is limited. We developed an ex vivo imaging approach in which fresh skin biopsies are mounted and then labeled with nanobodies or antibodies against cell surface markers on tissue-resident memory CD8.sup.+ T cells, other immune cells of interest, or extracellular tissue components. Subsequent longitudinal imaging allows one to describe the dynamic behavior of human skin-resident cells in situ. In addition, this strategy can be used to study immune cell function in murine skin. The ability to follow the spatiotemporal behavior of CD8.sup.+ T cells and other immune cells in skin, including their response to immune stimuli, provides a platform to investigate physiological immune cell behavior and immune cell behavior in skin diseases. The mounting, staining and imaging of skin samples requires ~1.5 d, and subsequent tracking analysis requires a minimum of 1 d. The optional production of fluorescently labeled nanobodies takes ~5 d. Human skin harbors various immune cells that are crucial for the control of injury and infection. However, the current understanding of immune cell function within viable human skin tissue is limited. We developed an ex vivo imaging approach in which fresh skin biopsies are mounted and then labeled with nanobodies or antibodies against cell surface markers on tissue-resident memory CD8+ T cells, other immune cells of interest, or extracellular tissue components. Subsequent longitudinal imaging allows one to describe the dynamic behavior of human skin-resident cells in situ. In addition, this strategy can be used to study immune cell function in murine skin. The ability to follow the spatiotemporal behavior of CD8+ T cells and other immune cells in skin, including their response to immune stimuli, provides a platform to investigate physiological immune cell behavior and immune cell behavior in skin diseases. The mounting, staining and imaging of skin samples requires ~1.5 d, and subsequent tracking analysis requires a minimum of 1 d. The optional production of fluorescently labeled nanobodies takes ~5 d.This protocol describes how to follow the behavior of immune cells by imaging ex vivo cultured human or mouse skin biopsies following labeling with antibody or nanobody reagents against specific cell surface markers. Human skin harbors various immune cells that are crucial for the control of injury and infection. However, the current understanding of immune cell function within viable human skin tissue is limited. We developed an ex vivo imaging approach in which fresh skin biopsies are mounted and then labeled with nanobodies or antibodies against cell surface markers on tissue-resident memory CD8+ T cells, other immune cells of interest, or extracellular tissue components. Subsequent longitudinal imaging allows one to describe the dynamic behavior of human skin-resident cells in situ. In addition, this strategy can be used to study immune cell function in murine skin. The ability to follow the spatiotemporal behavior of CD8+ T cells and other immune cells in skin, including their response to immune stimuli, provides a platform to investigate physiological immune cell behavior and immune cell behavior in skin diseases. The mounting, staining and imaging of skin samples requires ~1.5 d, and subsequent tracking analysis requires a minimum of 1 d. The optional production of fluorescently labeled nanobodies takes ~5 d.Human skin harbors various immune cells that are crucial for the control of injury and infection. However, the current understanding of immune cell function within viable human skin tissue is limited. We developed an ex vivo imaging approach in which fresh skin biopsies are mounted and then labeled with nanobodies or antibodies against cell surface markers on tissue-resident memory CD8+ T cells, other immune cells of interest, or extracellular tissue components. Subsequent longitudinal imaging allows one to describe the dynamic behavior of human skin-resident cells in situ. In addition, this strategy can be used to study immune cell function in murine skin. The ability to follow the spatiotemporal behavior of CD8+ T cells and other immune cells in skin, including their response to immune stimuli, provides a platform to investigate physiological immune cell behavior and immune cell behavior in skin diseases. The mounting, staining and imaging of skin samples requires ~1.5 d, and subsequent tracking analysis requires a minimum of 1 d. The optional production of fluorescently labeled nanobodies takes ~5 d. Human skin harbors various immune cells that are crucial for the control of injury and infection. However, the current understanding of immune cell function within viable human skin tissue is limited. We developed an ex vivo imaging approach in which fresh skin biopsies are mounted and then labeled with nanobodies or antibodies against cell surface markers on tissue-resident memory CD8 + T cells, other immune cells of interest, or extracellular tissue components. Subsequent longitudinal imaging allows one to describe the dynamic behavior of human skin-resident cells in situ. In addition, this strategy can be used to study immune cell function in murine skin. The ability to follow the spatiotemporal behavior of CD8 + T cells and other immune cells in skin, including their response to immune stimuli, provides a platform to investigate physiological immune cell behavior and immune cell behavior in skin diseases. The mounting, staining and imaging of skin samples requires ~1.5 d, and subsequent tracking analysis requires a minimum of 1 d. The optional production of fluorescently labeled nanobodies takes ~5 d. This protocol describes how to follow the behavior of immune cells by imaging ex vivo cultured human or mouse skin biopsies following labeling with antibody or nanobody reagents against specific cell surface markers. Human skin harbors various immune cells that are crucial for the control of injury and infection. However, the current understanding of immune cell function within viable human skin tissue is limited. We developed an ex vivo imaging approach in which fresh skin biopsies are mounted and then labeled with nanobodies or antibodies against cell surface markers on tissue-resident memory CD8 T cells, other immune cells of interest, or extracellular tissue components. Subsequent longitudinal imaging allows one to describe the dynamic behavior of human skin-resident cells in situ. In addition, this strategy can be used to study immune cell function in murine skin. The ability to follow the spatiotemporal behavior of CD8 T cells and other immune cells in skin, including their response to immune stimuli, provides a platform to investigate physiological immune cell behavior and immune cell behavior in skin diseases. The mounting, staining and imaging of skin samples requires ~1.5 d, and subsequent tracking analysis requires a minimum of 1 d. The optional production of fluorescently labeled nanobodies takes ~5 d. |
| Audience | Academic |
| Author | Luiten, Rosalie M. Mertz, Marjolijn Teunissen, Marcel B. M. Dijkgraaf, Feline E. Vredevoogd, David W. Hoogenboezem, Mark Matos, Tiago R. Schumacher, Ton N. Toebes, Mireille |
| Author_xml | – sequence: 1 givenname: Feline E. surname: Dijkgraaf fullname: Dijkgraaf, Feline E. organization: Division of Molecular Oncology and Immunology, Oncode Institute, The Netherlands Cancer Institute – sequence: 2 givenname: Mireille surname: Toebes fullname: Toebes, Mireille organization: Division of Molecular Oncology and Immunology, Oncode Institute, The Netherlands Cancer Institute – sequence: 3 givenname: Mark surname: Hoogenboezem fullname: Hoogenboezem, Mark organization: Research Facility, Sanquin Amsterdam – sequence: 4 givenname: Marjolijn orcidid: 0000-0002-9188-3579 surname: Mertz fullname: Mertz, Marjolijn organization: BioImaging Facility, The Netherlands Cancer Institute – sequence: 5 givenname: David W. surname: Vredevoogd fullname: Vredevoogd, David W. organization: Division of Molecular Oncology and Immunology, Oncode Institute, The Netherlands Cancer Institute – sequence: 6 givenname: Tiago R. orcidid: 0000-0003-2864-8207 surname: Matos fullname: Matos, Tiago R. organization: Department of Dermatology and Netherlands Institute for Pigment Disorders, Amsterdam University Medical Centers, University of Amsterdam – sequence: 7 givenname: Marcel B. M. orcidid: 0000-0003-1119-6662 surname: Teunissen fullname: Teunissen, Marcel B. M. organization: Department of Dermatology and Netherlands Institute for Pigment Disorders, Amsterdam University Medical Centers, University of Amsterdam – sequence: 8 givenname: Rosalie M. surname: Luiten fullname: Luiten, Rosalie M. organization: Department of Dermatology and Netherlands Institute for Pigment Disorders, Amsterdam University Medical Centers, University of Amsterdam – sequence: 9 givenname: Ton N. orcidid: 0000-0003-0517-8804 surname: Schumacher fullname: Schumacher, Ton N. email: t.schumacher@nki.nl organization: Division of Molecular Oncology and Immunology, Oncode Institute, The Netherlands Cancer Institute |
| BackLink | https://www.ncbi.nlm.nih.gov/pubmed/33349704$$D View this record in MEDLINE/PubMed |
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| Title | Labeling and tracking of immune cells in ex vivo human skin |
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