Pulse-shaped two-photon excitation of a fluorescent base analogue approaches single-molecule sensitivity
| Autor: | Rachel S. Fisher, Mattias Bood, Anders Foller Füchtbauer, Morten Grøtli, David Nobis, Marcus Wilhelmsson, Steven W. Magennis, Anita C. Jones |
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| Jazyk: | angličtina |
| Rok vydání: | 2018 |
| Předmět: |
Microscope
Materials science business.industry General Physics and Astronomy Quantum yield Context (language use) 010402 general chemistry 01 natural sciences Fluorescence Pulse shaping 0104 chemical sciences law.invention 010309 optics Two-photon excitation microscopy law 0103 physical sciences Optoelectronics Physical and Theoretical Chemistry business Ultrashort pulse Excitation |
| Zdroj: | Fisher, R S, Nobis, D, Füchtbauer, A F, Bood, M, Grøtli, M, Wilhelmsson, L M, Jones, A C & Magennis, S W 2018, ' Pulse-shaped two-photon excitation of a fluorescent base analogue approaches single-molecule sensitivity ', Physical Chemistry Chemical Physics . https://doi.org/10.1039/C8CP05496G |
| ISSN: | 1463-9076 |
| DOI: | 10.1039/C8CP05496G |
| Popis: | Fluorescent nucleobase analogues (FBAs) have many desirable features in comparison to extrinsic fluorescent labels, but they are yet to find application in ultrasensitive detection. Many of the disadvantages of FBAs arise from their short excitation wavelengths (often in the ultraviolet), making two-photon excitation a potentially attractive approach. Pentacyclic adenine (pA) is a recently developed FBA that has an exceptionally high two-photon brightness. We have studied the two-photon-excited fluorescence properties of pA and how they are affected by incorporation in DNA. We find that pA is more photostable under two-photon excitation than via resonant absorption. When incorporated in an oligonucleotide, pA has a high two-photon cross section and emission quantum yield, varying with sequence context, resulting in the highest reported brightness for such a probe. The use of a two-photon microscope with ultrafast excitation and pulse shaping has allowed the detection of pA-containing oligonucleotides in solution with a limit of detection of ∼5 molecules, demonstrating that practical single-molecule detection of FBAs is now within reach. |
| Databáze: | OpenAIRE |
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