Glass capillary systems for micro-volume fluorometry
•Reduction in consumption of florescent materials required for analysis is discussed.•Systems using cuvettes, capillaries and anti-resonant hollow core fibers are compared.•Performance of capillary-based system is optimized for green fluorescent protein (GFP).•Volumes and GFP concentration as low as...
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Veröffentlicht in: | Measurement : journal of the International Measurement Confederation 2025-01, Vol.240, p.115569, Article 115569 |
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Sprache: | eng |
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Zusammenfassung: | •Reduction in consumption of florescent materials required for analysis is discussed.•Systems using cuvettes, capillaries and anti-resonant hollow core fibers are compared.•Performance of capillary-based system is optimized for green fluorescent protein (GFP).•Volumes and GFP concentration as low as 2.9 μL and 1.5 nM were analysed, respectively.•The solution is versatile, cost-effective and makes minimization of wastes possible.
Capillary-based fluorometry can be considered a promising alternative to cuvette-based methods, enabling low-volume measurements while reducing analyte consumption and waste generation. In this work, we compared the performance of three types of glass container systems, i.e., anti-resonant hollow core fibers, capillaries, and standard cuvettes, using green fluorescent protein (GFP) as a reference analyte. Among these, when the volume of the analyte and the fluorescence intensity are assessed, a capillary accommodating 2.9 μL was found as an optimal choice. The capillary-based interrogation setup was further optimized to enhance sensitivity by adjusting the florescence-exciting laser beam position and optimizing the capillary’s shape. The obtained capillary-based setup achieved a noteworthy 85 % reduction in sample volume compared to previous studies utilizing capillary-based fluorometry, highlighting its significant contribution to material conservation. The system achieved a limit of detection as low as 26 μg/mL (1.01 nM) GFP concentration and a resolution of 3.5 μg/mL. With its low cost and potential for reusability, capillary-based fluorometry presents a compelling alternative to traditional cuvette-based fluorometry, offering a promising solution for micro-volume analysis. |
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ISSN: | 0263-2241 |
DOI: | 10.1016/j.measurement.2024.115569 |