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PubMed · 2630641

[Fluoromicroscopy].

J F Peli
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Abstract

The fluorescence is a photoluminescent phenomena consisting in the emission of light under the influence of a light wave excitation. Two types of fluorescence exist: the primary fluorescence (natural) and the secondary (induced by a fluorochrome) The fluoromicroscope is composed of two parts. One of which, unspecific, consists in the classical optical system of the photonic microscopy. The other part, specific, consists in a source of light at a high mercurial pressure and a double filtration system of excitation and pause. There are many applications of this technique. In odontology, it is frequently used for the study of apposition in calcified tissues after their labelling by injection of a fluorochrome.

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Fluorescent DyesMicroscopy, FluorescenceEnglish AbstractJournal Article

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BibTeXRIS

J F Peli. 1989. [Fluoromicroscopy].. https://pubmed.ncbi.nlm.nih.gov/2630641/

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IMPACT OF FLUORESCENT DYES ON MUTATIONS IN NEXT GENERATION SEQUENCING LIBRARY GENERATION.

DNA labelling fluorescent dyes such as ethidium bromide have long been considered to be highly mutagenic during DNA replication. While recent studies have pushed back on this narrative, the intercalative nature of these dyes continues to raise the possibility that these dyes can induce mutations. The iconPCR instrument by n6tec uses fluorescent dyes to measure amplification in real time and to adjust cycling conditions. However, since this use of qPCR is preparative and not analytical, mutations introduced by fluorescent dyes would be propagated into the sequencing reaction. To address the impact of these dyes on downstream analyses, we have performed routine mutation calling as well as mutational signature analysis on samples amplified using the iconPCR in the presence of either SYBR or EvaGreen. Sequence analysis revealed very minimal impacts of dyes on the reactions, largely within the noise regimen with only subtle changes in mutation rates seen. Mutational signature analysis was unable to identify any key signatures assignable to the dyes in either substitutions or indel domains. The mutational impact of intercalating dyes during fluorescence-guided amplification is therefore minimal and can be disregarded in all but the most sensitive NGS applications.

Fluorescent Dyes↗