Novel PE and APC tandems: Additional near‐infrared fluorochromes for use in spectral flow cytometry. Issue 10 (2nd February 2022)

• Record Type: Journal Article
• Authors: Seong, Yekyung; Nguyen, Denny X; Wu, Yian; Thakur, Archana; Harding, Fiona; Nguyen, Tuan Andrew
• Other Names: Lannigan Joanne guestEditor.
• Is Part Of: Cytometry. Volume 101:Issue 10(2022)
• Page Start: 835
• Page End: 845
• Publication Date: 2022-02-02
• Subjects: flow cytometry -- fluorochrome -- high content -- high dimensional -- near infrared -- spectral; Flow cytometry -- Periodicals; Imaging systems in biology -- Periodicals; Imaging systems in medicine -- Periodicals; Diagnostic imaging -- Periodicals; 571.605
• DOI: 10.1002/cyto.a.24537
• Languages: English
• ISSNs: 1552-4922

Abstract

Abstract: 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.