Optimization in Detection of Antigen‐Specific T Cells Through Differentially Labeled MHC Multimers
A large variety of fluorescent molecules are used on a regular basis to tag major histocompatibility complex (MHC) multimers for detection of antigen‐specific T cells. We have evaluated the way in which the choice of fluorescent label can impact the detection of MHC multimer binding T cells in an ex...
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| Veröffentlicht in: | Cytometry. Part A 2020-09, Vol.97 (9), p.955-964 |
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| container_title | Cytometry. Part A |
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| creator | Pedersen, Natasja Wulff Laske, Karoline Maurer, Dominik Welters, Marij Walter, Steffen Gouttefangeas, Cécile Hadrup, Sine Reker |
| description | A large variety of fluorescent molecules are used on a regular basis to tag major histocompatibility complex (MHC) multimers for detection of antigen‐specific T cells. We have evaluated the way in which the choice of fluorescent label can impact the detection of MHC multimer binding T cells in an exploratory proficiency panel where detection of MHC multimer binding T cells was assessed across 16 different laboratories. We found that the staining index (SI) of the multimer reagent provided the best direct correlation with the value of a given fluorochrome for T cell detection studies. The SI is dependent on flow cytometer settings and chosen antibody panel; hence, the optimal fluorochrome selection may differ from lab to lab. Consequently, we describe a strategy to evaluate performance of the detection channels and optimize the SI for selected fluorescent molecules. This approach can easily be used to test and optimize fluorescence detection in relation to MHC multimer staining and in general, for antibody‐based identification of rare cell populations. © 2019 The Authors. Cytometry Part A published by Wiley Periodicals, Inc. on behalf of International Society for Advancement of Cytometry. |
| doi_str_mv | 10.1002/cyto.a.23942 |
| format | Article |
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Part A</title><addtitle>Cytometry A</addtitle><description>A large variety of fluorescent molecules are used on a regular basis to tag major histocompatibility complex (MHC) multimers for detection of antigen‐specific T cells. We have evaluated the way in which the choice of fluorescent label can impact the detection of MHC multimer binding T cells in an exploratory proficiency panel where detection of MHC multimer binding T cells was assessed across 16 different laboratories. We found that the staining index (SI) of the multimer reagent provided the best direct correlation with the value of a given fluorochrome for T cell detection studies. The SI is dependent on flow cytometer settings and chosen antibody panel; hence, the optimal fluorochrome selection may differ from lab to lab. Consequently, we describe a strategy to evaluate performance of the detection channels and optimize the SI for selected fluorescent molecules. This approach can easily be used to test and optimize fluorescence detection in relation to MHC multimer staining and in general, for antibody‐based identification of rare cell populations. © 2019 The Authors. Cytometry Part A published by Wiley Periodicals, Inc. on behalf of International Society for Advancement of Cytometry.</description><subject>Antibodies</subject><subject>antigen specific CD8 T cells</subject><subject>Antigens</subject><subject>Binding</subject><subject>Cytometry</subject><subject>flow cytometry</subject><subject>Fluorescence</subject><subject>Lymphocytes</subject><subject>Lymphocytes T</subject><subject>Major histocompatibility complex</subject><subject>MHC multimers</subject><subject>Optimization</subject><subject>optimized detection of fluorescence</subject><subject>Original</subject><subject>Performance evaluation</subject><subject>Reagents</subject><subject>Staining</subject><issn>1552-4922</issn><issn>1552-4930</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2020</creationdate><recordtype>article</recordtype><sourceid>24P</sourceid><sourceid>WIN</sourceid><recordid>eNp9kc1uEzEURkcIREthxxpZYsOiSf07M94gRdPSVkqVBWHByvJ4rhNXzjjYM6Cw4hF4Rp4EtylRYcHKtu7R8f30FcVrgqcEY3pmdkOY6illktMnxTERgk64ZPjp4U7pUfEipVuMmcCMPi-OGKlxLaU8LrrFdnAb910PLvTI9egcBjD3j2DRrB_cCvpfP35-3IJx1hm0RA14n9ByHcO4WqNzZy1EyKD2fofmugUPHbq5atDN6LMbYnpZPLPaJ3j1cJ4Unz5cLJuryXxxed3M5hPDCacTI-uOdayWpWVYGxClNqYVAixpgeGSdLy0EltDO4Fty2XJpIWKSIHLkvKOnRTv997t2G6gM3mrqL3aRrfRcaeCdurvSe_WahW-qkpwXNZ1Frx7EMTwZYQ0qI1LJufVPYQxKcoorUT-l2T07T_obRhjn-MpyjmuK1JJnqnTPWViSCmCPSxDsLqrT93Vp7S6ry_jbx4HOMB_-soA3wPfnIfdf2Wq-bxczPbe34zfqTY</recordid><startdate>202009</startdate><enddate>202009</enddate><creator>Pedersen, Natasja Wulff</creator><creator>Laske, Karoline</creator><creator>Maurer, Dominik</creator><creator>Welters, Marij</creator><creator>Walter, Steffen</creator><creator>Gouttefangeas, Cécile</creator><creator>Hadrup, Sine Reker</creator><general>John Wiley & Sons, Inc</general><general>Wiley Subscription Services, Inc</general><scope>24P</scope><scope>WIN</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QO</scope><scope>7TK</scope><scope>8FD</scope><scope>FR3</scope><scope>P64</scope><scope>7X8</scope><scope>5PM</scope><orcidid>https://orcid.org/0000-0002-7252-4562</orcidid></search><sort><creationdate>202009</creationdate><title>Optimization in Detection of Antigen‐Specific T Cells Through Differentially Labeled MHC Multimers</title><author>Pedersen, Natasja Wulff ; Laske, Karoline ; Maurer, Dominik ; Welters, Marij ; Walter, Steffen ; Gouttefangeas, Cécile ; Hadrup, Sine Reker</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c4142-c98d3d3896f30ace56accb55ef1be3061d46f90fc2d50fb49639fe719506624d3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2020</creationdate><topic>Antibodies</topic><topic>antigen specific CD8 T cells</topic><topic>Antigens</topic><topic>Binding</topic><topic>Cytometry</topic><topic>flow cytometry</topic><topic>Fluorescence</topic><topic>Lymphocytes</topic><topic>Lymphocytes T</topic><topic>Major histocompatibility complex</topic><topic>MHC multimers</topic><topic>Optimization</topic><topic>optimized detection of fluorescence</topic><topic>Original</topic><topic>Performance evaluation</topic><topic>Reagents</topic><topic>Staining</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Pedersen, Natasja Wulff</creatorcontrib><creatorcontrib>Laske, Karoline</creatorcontrib><creatorcontrib>Maurer, Dominik</creatorcontrib><creatorcontrib>Welters, Marij</creatorcontrib><creatorcontrib>Walter, Steffen</creatorcontrib><creatorcontrib>Gouttefangeas, Cécile</creatorcontrib><creatorcontrib>Hadrup, Sine Reker</creatorcontrib><collection>Wiley Online Library Open Access</collection><collection>Wiley Free Content</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Biotechnology Research Abstracts</collection><collection>Neurosciences Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Cytometry. 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| subjects | Antibodies antigen specific CD8 T cells Antigens Binding Cytometry flow cytometry Fluorescence Lymphocytes Lymphocytes T Major histocompatibility complex MHC multimers Optimization optimized detection of fluorescence Original Performance evaluation Reagents Staining |
| title | Optimization in Detection of Antigen‐Specific T Cells Through Differentially Labeled MHC Multimers |
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