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Novel PE and APC tandems: Additional near‐infrared fluorochromes for use in spectral flow cytometry

Title: Novel PE and APC tandems: Additional near‐infrared fluorochromes for use in spectral flow cytometry
Authors: Seong, Yekyung; Nguyen, Denny X; Wu, Yian; Thakur, Archana; Harding, Fiona; Nguyen, Tuan Andrew
Contributors: AbbVie Biotherapeutics
Source: Cytometry Part A ; volume 101, issue 10, page 835-845 ; ISSN 1552-4922 1552-4930
Publisher Information: Wiley
Publication Year: 2022
Collection: Wiley Online Library (Open Access Articles via Crossref)
Description: Recent advances in flow cytometry instrumentation and fluorochrome chemistries have greatly increased fluorescent conjugated antibody combinations that can be used reliably and easily in routine experiments. The Cytek Aurora flow cytometer was first released with three excitation lasers (405, 488, and 640 nm) and incorporated the latest Avalanche Photodiode (APD) technology, demonstrating significant improvement in sensitivity for fluorescent emission signals longer than 800 nm. However, there are limited commercially available fluorochromes capable of excitation with peak emission signals beyond 800 nm. To address this gap, we engineered six new fluorochromes: PE‐750, PE‐800, PE‐830 for the 488 nm laser and APC‐750, APC‐800, APC‐830 for the 640 nm laser. Utilizing the principal of fluorescence resonance energy transfer (FRET), these novel structures were created by covalently linking a protein donor dye with an organic small molecule acceptor dye. Additionally, each of these fluorochrome conjugates were shown to be compatible with fixation/permeabilization buffer reagents, and demonstrated acceptable brightness and stability when conjugated to antigen‐specific monoclonal antibodies. These six novel fluorochrome reagents can increase the numbers of fluorochromes that can be used on a spectral flow cytometer.
Document Type: article in journal/newspaper
Language: English
DOI: 10.1002/cyto.a.24537
Availability: http://dx.doi.org/10.1002/cyto.a.24537; https://onlinelibrary.wiley.com/doi/pdf/10.1002/cyto.a.24537; https://onlinelibrary.wiley.com/doi/full-xml/10.1002/cyto.a.24537
Rights: http://creativecommons.org/licenses/by-nc-nd/4.0/
Accession Number: edsbas.7B309FB3
Database: BASE