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Fluorophores and fluorescence detection are central to many research applications. Many fixed and live cell imaging techniques rely on immunofluorescence or fluorescent proteins. Fluorescence-based techniques can also be used in many common laboratory assays, including flow cytometry, ELISA, western blot, immunohistochemistry (IHC) and immunocytochemistry (ICC).Â
Fluorescent detection methods have several advantages over other analytical techniques, including high sensitivity, quantitative analysis, and the potential to carry out multiplex analysis using a range of available lasers, optical filters, and fluorophores.Â
This guide will enhance your understanding of how to use fluorescence in your research and advise you on choosing the right fluorophores and fluorescent proteins for your experiments. It begins with a basic introduction to fluorescence and how it works, followed by discussions of more advanced fluorescence mechanisms, such as fluorescence resonance energy transfer (FRET) and time-resolved fluorescence (TRF). Finally, the guide covers essential reagents for fluorescence-based experimentation, including fluorophores, tandem dyes, and fluorescent proteins.Â
Fluorescence is a light signal detected when a chemical compound called a fluorophore absorbs energy at a specific wavelength, causing it to become excited.
Fluorophores or fluorochromes are photoreactive chemicals that absorb and emit energy in a predictable fashion.
Time-resolved fluorescence methods can overcome the traditional limitations of conventional fluorophores.
In fluorescence resonance energy transfer (FRET), a donor fluorophore is excited by a light source and transfers its energy to a nearby acceptor fluorophore.
Selecting the right fluorophore can be difficult, so our experts have collected together all the consideration you may need to make when making your decision.