Time-resolved and two-photon emission imaging microscopy of live cells with inert platinum complexes

Botchway, Stanley W and Charnley, Mirren and Haycock, John W and Parker, Anthony W and Rochester, David L and Weinstein, Julia A and Williams, J A Gareth (2008) Time-resolved and two-photon emission imaging microscopy of live cells with inert platinum complexes. Proceedings of the National Academy of Sciences of the United States of America, 105 (42). pp. 16071-16076. ISSN 0027-8424

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Abstract

This work explores time-resolved emission imaging microscopy (TREM) for noninvasive imaging and mapping of live cells on a hitherto uncharted microsecond time scale. Simple robust molecules for this purpose have long been sought. We have developed highly emissive, synthetically versatile, and photostable platinum(II) complexes that make TREM a practicable reality. [PtLCl], {HL = 1,3-di(2-pyridyl)benzene and derivatives}, are charge-neutral, small molecules that have low cytotoxicity and accumulate intracellularly within a remarkably short incubation time of 5 min, apparently under diffusion control. Their microsecond lifetimes and emission quantum yields of up to 70% are exceptionally high for transition metal complexes and permit the application of TREM to be demonstrated in a range of live cell types-normal human dermal fibroblast, neoplastic C8161 and CHO cells. [PtLCl] are thus likely to be suitable emission labels for any eukaryotic cell types. The high photostability of [PtLCl] under intense prolonged irradiation has allowed the development of tissue-friendly NIR two-photon excitation (TPE) in conjunction with transition metal complexes in live cells. A combination of confocal one-photon excitation, nonlinear TPE, and microsecond time-resolved imaging has revealed (i) preferential localization of the complexes to intracellular nucleic acid structures, in particular the nucleoli and (ii) the possibility of measuring intracellular emission lifetimes in the microsecond range. The combination of TREM, TPE, and Pt(II) complexes will be a powerful tool for investigating intracellular processes in vivo, because the long lifetimes allow discrimination from autofluorescence and open up the use of commonplace technology.

Item Type:
Journal Article
Journal or Publication Title:
Proceedings of the National Academy of Sciences of the United States of America
Uncontrolled Keywords:
/dk/atira/pure/subjectarea/asjc/1000
Subjects:
ID Code:
132508
Deposited By:
Deposited On:
08 Apr 2019 10:40
Refereed?:
Yes
Published?:
Published
Last Modified:
28 Sep 2020 03:00