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Proceedings Paper

Two-photon phosphorescence lifetime microscopy (2PLM) for high resolution imaging of oxygen
Author(s): Louise E. Sinks; Emmanuel Roussakis; Sava Sakadžic; Gregory P. Robbins; Daniel A. Hammer; Anna Devor; David A. Boas; Sergei A. Vinogradov
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Paper Abstract

We present a new method for imaging of oxygen in biological samples with micron-scale resolution and threedimensional capability. Our technique combines phosphorescence quenching with multiphoton laser scanning microscopy. It is termed two-photon phosphorescence lifetime microscopy (2PLM). 2PLM is made possible due to specially designed dendritic probes, in which phosphorescence of metalloporphyrins upon two-photon excitation is enhanced by intramolecular Förster-type energy transfer from two-photon antennae chromophores, located in the same probe molecule. Oxygen sensitivity of the probes is regulated by dendritic encapsulation of the core metalloporphyrin, while peripheral PEG groups on the dendrimer isolate the probes from contacts with biological macromolecules (e.g. proteins, nucleic acids etc) in the environment. Instrumentation and resolution of the method are discussed along with approaches to intracellular delivery of the probes. 2PLM was validated in pilot imaging experiments and used to obtain depth-resolved high-resolution oxygen maps in vivo in the brain.

Paper Details

Date Published: 11 February 2011
PDF: 8 pages
Proc. SPIE 7903, Multiphoton Microscopy in the Biomedical Sciences XI, 79032A (11 February 2011); doi: 10.1117/12.873650
Show Author Affiliations
Louise E. Sinks, Univ. of Pennsylvania (United States)
Emmanuel Roussakis, Univ. of Pennsylvania (United States)
Sava Sakadžic, Massachusetts General Hospital (United States)
Gregory P. Robbins, Univ. of Pennsylvania (United States)
Daniel A. Hammer, Univ. of Pennsylvania (United States)
Anna Devor, Univ. of California, San Diego (United States)
David A. Boas, Massachusetts General Hospital (United States)
Sergei A. Vinogradov, Univ. of Pennsylvania (United States)


Published in SPIE Proceedings Vol. 7903:
Multiphoton Microscopy in the Biomedical Sciences XI
Ammasi Periasamy; Karsten König; Peter T. C. So, Editor(s)

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