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Cellular localisation of structurally diverse diphenylacetylene fluorophores

Chisholm, David R.; Hughes, Joshua G.; Blacker, Thomas S.; Humann, Rachel; Adams, Candace; Callaghan, Daniel; Pujol, Alba; Lembicz, Nicola K.; Bain, Angus J.; Girkin, John M.; Ambler, Carrie A.; Whiting, Andrew


Josh Hughes
PGR Student Doctor of Philosophy

Thomas S. Blacker

Rachel Humann

Candace Adams

Daniel Callaghan

Alba Pujol

Nicola K. Lembicz

Angus J. Bain


Fluorescent probes are increasingly used as reporter molecules in a wide variety of biophysical experiments, but when designing new compounds it can often be difficult to anticipate the effect that changing chemical structure can have on cellular localisation and fluorescence behaviour. To provide further chemical rationale for probe design, a series of donor–acceptor diphenylacetylene fluorophores with varying lipophilicities and structures were synthesised and analysed in human epidermal cells using a range of cellular imaging techniques. These experiments showed that, within this family, the greatest determinants of cellular localisation were overall lipophilicity and the presence of ionisable groups. Indeed, compounds with high log D values (>5) were found to localise in lipid droplets, but conversion of their ester acceptor groups to the corresponding carboxylic acids caused a pronounced shift to localisation in the endoplasmic reticulum. Mildly lipophilic compounds (log D = 2–3) with strongly basic amine groups were shown to be confined to lysosomes i.e. an acidic cellular compartment, but sequestering this positively charged motif as an amide resulted in a significant change to cytoplasmic and membrane localisation. Finally, specific organelles including the mitochondria could be targeted by incorporating groups such as a triphenylphosphonium moiety. Taken together, this account illustrates a range of guiding principles that can inform the design of other fluorescent molecules but, moreover, has demonstrated that many of these diphenylacetylenes have significant utility as probes in a range of cellular imaging studies.


Chisholm, D. R., Hughes, J. G., Blacker, T. S., Humann, R., Adams, C., Callaghan, D., …Whiting, A. (2020). Cellular localisation of structurally diverse diphenylacetylene fluorophores. Organic and Biomolecular Chemistry, 18(45), 9231-9245.

Journal Article Type Article
Acceptance Date Sep 15, 2020
Online Publication Date Sep 17, 2020
Publication Date 2020
Deposit Date Mar 2, 2021
Journal Organic and Biomolecular Chemistry
Print ISSN 1477-0520
Electronic ISSN 1477-0539
Publisher Royal Society of Chemistry
Volume 18
Issue 45
Pages 9231-9245
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