Infrared-LAMP: two-photon uncaging and imaging of gap junctional communication in three dimensions

Dakin, Kenneth; Li, Wen Hong

doi:10.1038/nmeth1206-959

Cited by 38 publications

(34 citation statements)

References 6 publications

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“…elegans embryonic cells, we used a photoactivatable (caged) fluorescent dye and the uncaging/imaging technique (Dakin et al, 2005;Dakin and Li, 2006). Embryonic neurons were isolated from nsy-5p::mCherry transgenic animals, in which neurons of the NSY-5 network were labeled with mCherry (Shaner et al, 2004;Shaner et al, 2005).…”

Section: Nsy-5 Gap Junctions Mediate Small Molecule Transfer Between mentioning

confidence: 99%

Intercellular calcium signaling in a gap junction-coupled cell network establishes asymmetric neuronal fates in C. elegans

et al. 2012

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Section: Nsy-5 Gap Junctions Mediate Small Molecule Transfer Between mentioning

confidence: 99%

Intercellular calcium signaling in a gap junction-coupled cell network establishes asymmetric neuronal fates in C. elegans

et al. 2012

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“…As a result, these functional fluorescent probes permit the implementation of imaging and spectroscopic protocols that are otherwise inaccessible with conventional fluorophores. Indeed, photoactivatable fluorophores are routinely used as calibration standards in photolysis experiments [131][132][133], to probe the activity and dynamics of biomolecular systems [48,49,55,61,62,68,71,[134][135][136][137][138][139][140][141][142], to monitor the flow dynamics of fluids [143][144][145][146][147][148][149][150][151][152], and to reconstruct subdiffraction images [57-59, 74-76, 98, 99, 103, 104, 129, 153-160]. The common theme of all these strategies is the interplay of activating and exciting beams to switch fluorescence on within a defined region of space at a given interval of time.…”

Section: Applications Of Photoactivatable Fluorophoresmentioning

confidence: 99%

“…Sequences of images ( Figures 9(a)-9(c)), recorded upon irradiation at λ Ex over the course of several seconds, reveal clearly a change in the spatial distribution of the activated fluorescence that is indicative of microfilament dynamics. In fact, similar imaging protocols have been extended to the investigation of the dynamics of a diversity of biomolecular systems [134,135,[138][139][140][141][142]. In addition to the diffusion of individual molecules within a media of interest, similar configurations can be adapted to the investigation of fluid dynamics [143][144][145][146][147][148][149][150][151][152].…”

Section: Applications Of Photoactivatable Fluorophoresmentioning

confidence: 99%

Photoactivatable Fluorophores

Raymo

2012

ISRN Physical Chemistry

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Photoactivatable fluorophores switch from a nonemissive to an emissive state upon illumination at an activating wavelength and then emit after irradiation at an exciting wavelength. The interplay of such activation and excitation events can be exploited to switch fluorescence on in a defined region of space at a given interval of time. In turn, the spatiotemporal control of fluorescence translates into the opportunity to implement imaging and spectroscopic schemes that are not possible with conventional fluorophores. Specifically, photoactivatable fluorophores permit the monitoring of dynamic processes in real time as well as the reconstruction of images with subdiffraction resolution. These promising applications can have a significant impact on the characterization of the structures and functions of biomolecular systems. As a result, strategies to implement mechanisms for fluorescence photoactivation with synthetic fluorophores are particularly valuable. In fact, a number of versatile operating principles have already been identified to activate the fluorescence of numerous members of the main families of synthetic dyes. These methods are based on either the irreversible cleavage of covalent bonds or the reversible opening and closing of rings. This paper overviews the fundamental mechanisms that govern the behavior of these photoresponsive systems, illustrates structural designs for fluorescence photoactivation, and provides representative examples of photoactivatable fluorophores in actions.

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“…In one such study, dissected pancreatic acini were loaded with the green fluorescent dye calcein/AM (Fig. 5), to label individual acinar cells, and the cell membrane permeable caged coumarin dye NPE-HCC2.AM (Dakin & Li, 2006). The caged coumarin is not fluorescent and cannot cross gap junctional channels.…”

Section: Cardiovascular Applications Of Tpe Microscopymentioning

confidence: 99%

Cardiovascular Imaging Using Two-Photon Microscopy

Scherschel

Rubart

2008

Microsc Microanal

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Two-photon excitation microscopy has become the standard technique for high resolution deep tissue and intravital imaging. It provides intrinsic three-dimensional resolution in combination with increased penetration depth compared to single-photon confocal microscopy. This article will describe the basic physical principles of two-photon excitation and will review its multiple applications to cardiovascular imaging, including second harmonic generation and fluorescence laser scanning microscopy. In particular, the capability and limitations of multiphoton microscopy to assess functional heterogeneity on a cellular scale deep within intact, Langendorff-perfused hearts are demonstrated. It will also discuss the use of two-photon excitation-induced release of caged compounds for the study of intracellular calcium signaling and intercellular dye transfer.

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Infrared-LAMP: two-photon uncaging and imaging of gap junctional communication in three dimensions

Cited by 38 publications

References 6 publications

Intercellular calcium signaling in a gap junction-coupled cell network establishes asymmetric neuronal fates in C. elegans

Intercellular calcium signaling in a gap junction-coupled cell network establishes asymmetric neuronal fates in C. elegans

Photoactivatable Fluorophores

Cardiovascular Imaging Using Two-Photon Microscopy

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