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Review
. 2018 Aug 14;8(51):29051-29061.
doi: 10.1039/c8ra02297f.

Small-molecule fluorescent probes and their design

Yanhua Fu  1 , Nathaniel S Finney  1
Affiliations
Review

Small-molecule fluorescent probes and their design

Yanhua Fu et al. RSC Adv. .

Abstract

Small-molecule fluorescent probes have become powerful tools for using light to advance the study of cell biology, discover new drugs, detect environmental contaminants, and further the detection of cancer. These applications correlate with the expansion of the fluorescent probe research community - small in the late 20th century, now a collection of more than a hundred research groups world-wide. This expansion required the entry of adventurous scientists from many other fields. This tutorial review introduces some important concepts related to fluorescent probe development. It is hoped that it will facilitate further expansion of the field by demystifying it.

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Conflict of interest statement

There are no conflicts to declare.

Figures

Fig. 1
Fig. 1. Simple scheme for excitation and emission in conjugated π systems.a,b,c aAs a matter of convention, excited states are usually denoted with an asterisk(*). Absorbed or emitted light is routinely abbreviated as hν, where ν denotes frequency, and hν gives the energy of the radiation. bThe initial excited state is formed with the electrons still spin-paired. cTerminology for ground states and excited states is deliberately minimized in this review. For more comprehensive discussion, see ref. 2.
Fig. 2
Fig. 2. Photoinduced electron transfer (PET) quenching by a donor (D).
Fig. 3
Fig. 3. A PET probe for trace cellular Cu2+. a Structure of Cu2+ coordination complex not shown in detail.
Fig. 4
Fig. 4. A PET probe for cellular hypochlorite based on oxidation of a pyrrole.
Fig. 5
Fig. 5. Increase in dipole moment upon excitation to the excited state.
Fig. 6
Fig. 6. Energy diagram correlating ICT with solvation, in polar solvents.
Fig. 7
Fig. 7. A pioneering ICT-based probe for cellular Ca2+ imaging, fura-2.
Fig. 8
Fig. 8. An ICT-based probe for H2S in cells.
Fig. 9
Fig. 9. Energy diagram for FRET. aThe double-headed arrow denotes the energy-matching of the lowest energy vibrational state of the donor excited state with a high energy vibrational level of the acceptor excited state. This is equivalent to the absorption/emission overlap shown in Fig. 10.
Fig. 10
Fig. 10. Emission overlap vs. λmax separation in a hypothetical FRET pair.
Fig. 11
Fig. 11. FRET-based probe for measurement of pH in lysosomes.
Fig. 12
Fig. 12. FRET-based probe for H2S detection in cells.
Fig. 13
Fig. 13. Representative molecules from some important fluorophore classes.
Fig. 14
Fig. 14. Interplay between emissive open-form and non-fluorescent spirocyclic closed-form of fluorescein and rhodamine derivatives.
Fig. 15
Fig. 15. Visual summary of how substituents in some important fluorophore classes can be modified to alter properties and function.
See this image and copyright information in PMC

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