Novel full‐spectral flow cytometry with multiple spectrally‐adjacent fluorescent proteins and fluorochromes and visualization of in vivo cellular movement
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- Koji Futamura
- FCM Business Department Life Science Business Division Medical Business Unit Sony Corporation Minato‐Ku Tokyo 108‐0075 Japan
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- Masashi Sekino
- Concept Development Department Application Technology Development Division System R&D Group, RDS Platform, Sony Corporation Shinagawa‐Ku Tokyo 141‐0001 Japan
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- Akihiro Hata
- Center for Innovation in Immunoregulative Technology and Therapeutics, Kyoto University Graduate School of Medicine Yoshida‐Konoe Kyoto 606‐8501 Japan
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- Ryoyo Ikebuchi
- Center for Innovation in Immunoregulative Technology and Therapeutics, Kyoto University Graduate School of Medicine Yoshida‐Konoe Kyoto 606‐8501 Japan
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- Yasutaka Nakanishi
- Center for Innovation in Immunoregulative Technology and Therapeutics, Kyoto University Graduate School of Medicine Yoshida‐Konoe Kyoto 606‐8501 Japan
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- Gyohei Egawa
- Department of Dermatology Kyoto University Graduate School of Medicine Kyoto 606‐8501 Japan
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- Kenji Kabashima
- Department of Dermatology Kyoto University Graduate School of Medicine Kyoto 606‐8501 Japan
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- Takeshi Watanabe
- Center for Innovation in Immunoregulative Technology and Therapeutics, Kyoto University Graduate School of Medicine Yoshida‐Konoe Kyoto 606‐8501 Japan
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- Motohiro Furuki
- FCM Business Department Life Science Business Division Medical Business Unit Sony Corporation Minato‐Ku Tokyo 108‐0075 Japan
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- Michio Tomura
- Center for Innovation in Immunoregulative Technology and Therapeutics, Kyoto University Graduate School of Medicine Yoshida‐Konoe Kyoto 606‐8501 Japan
書誌事項
- 公開日
- 2015-07-28
- 資源種別
- journal article
- 権利情報
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- http://creativecommons.org/licenses/by/4.0/
- DOI
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- 10.1002/cyto.a.22725
- 公開者
- Wiley
この論文をさがす
説明
<jats:title>Abstract</jats:title><jats:p>Flow cytometric analysis with multicolor fluoroprobes is an essential method for detecting biological signatures of cells. Here, we present a new full‐spectral flow cytometer (spectral‐FCM). Unlike conventional flow cytometer, this spectral‐FCM acquires the emitted fluorescence for all probes across the full‐spectrum from each cell with 32 channels sequential PMT unit after dispersion with prism, and extracts the signals of each fluoroprobe based on the spectral shape of each fluoroprobe using unique algorithm in high speed, high sensitive, accurate, automatic and real‐time. The spectral‐FCM detects the continuous changes in emission spectra from green to red of the photoconvertible protein, KikGR with high‐spectral resolution and separates spectrally‐adjacent fluoroprobes, such as FITC (Emission peak (Em) 519 nm) and EGFP (Em 507 nm). Moreover, the spectral‐FCM can measure and subtract autofluorescence of each cell providing increased signal‐to‐noise ratios and improved resolution of dim samples, which leads to a transformative technology for investigation of single cell state and function. These advances make it possible to perform 11‐color fluorescence analysis to visualize movement of multilinage immune cells by using KikGR‐expressing mice. Thus, the novel spectral flow cytometry improves the combinational use of spectrally‐adjacent various FPs and multicolor fluorochromes in metabolically active cell for the investigation of not only the immune system but also other research and clinical fields of use. © 2015 The Authors. Cytometry Part A Published by Wiley Periodicals, Inc. on behalf of ISAC</jats:p>
収録刊行物
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- Cytometry Part A
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Cytometry Part A 87 (9), 830-842, 2015-07-28
Wiley