Miltenyi Biotec BV & Co. KG, Department Chemical Biology, Friedrich-Ebert-Str. 68, 51429, Bergisch Gladbach, Germany.
DFG Cluster of Excellence livMatS @FIT and Albert-Ludwigs-Universität Freiburg, Institut für Organische Chemie, Albertstraße 21, 79104, Freiburg, Germany.
Angew Chem Int Ed Engl. 2024 May 13;63(20):e202402616. doi: 10.1002/anie.202402616. Epub 2024 Apr 2.
The application of spectrally unique, bright, and water-soluble fluorescent dyes is indispensable for the analysis of biological systems. Multiparameter flow cytometry is a powerful tool for characterization of mixed cell populations. To discriminate the different cell populations, they are typically stained by a set of fluorescent reagents, e.g., antibody-fluorophore conjugates. The number of parameters which can be studied simultaneously strongly depends on the availability of reagents which can be differentiated by their spectral properties. In this study a series of fluorescent polymer dyes was developed, that can be excited with a single violet laser (405 nm) but distinguished by their unique emission spectra. The polyfluorene-based polymers can be used on their own, or in combination with covalently bound small-molecule dyes to generate energy transfer constructs to red-shift the emission wavelength based on Förster resonance energy transfer (FRET). The polymer dyes were utilized in a biological flow cytometry assay by conjugating several of them to antibodies, demonstrating their effectiveness as reagents. This report represents the first systematic investigation of structure-property relationships for this type of fluorescent dyes.
光谱独特、明亮且水溶性好的荧光染料的应用对于生物系统的分析是不可或缺的。多参数流式细胞术是分析混合细胞群体的有力工具。为了区分不同的细胞群体，通常使用一组荧光试剂对其进行染色，例如抗体-荧光染料缀合物。可同时研究的参数数量强烈依赖于试剂的可用性，这些试剂可通过其光谱特性进行区分。在这项研究中，开发了一系列荧光聚合物染料，它们可以用单个紫光激光（405nm）激发，但可以通过其独特的发射光谱来区分。基于聚芴的聚合物可以单独使用，也可以与共价键合的小分子染料结合使用，以基于Förster 共振能量转移（FRET）产生能量转移构建体，将发射波长红移。通过将其中几种与抗体缀合，将聚合物染料用于生物流式细胞术测定中，证明了它们作为试剂的有效性。本报告代表了对这种类型的荧光染料的结构-性质关系的首次系统研究。
