Confocal fluorescence lifetime imaging of pH in single cells

1994 ◽  
Author(s):  
Renata Sanders ◽  
Hans C. Gerritsen ◽  
Arie Draaijer ◽  
P. M. Houpt ◽  
Sjaak J. F. van Veen ◽  
...  
Keyword(s):  
Fluorescence Lifetime ◽  
Single Cells ◽  
Fluorescence Lifetime Imaging ◽  
Lifetime Imaging ◽  
Confocal Fluorescence

Confocal fluorescence lifetime imaging of free calcium in single cells

1994 ◽  
Vol 4 (4) ◽  
pp. 291-294 ◽  
Author(s):  
Renata Sanders ◽  
Hans C. Gerritsen ◽  
Arie Draaijer ◽  
Piet M. Houpt ◽  
Yehudi K. Levine
Keyword(s):  
Fluorescence Lifetime ◽  
Single Cells ◽  
Free Calcium ◽  
Fluorescence Lifetime Imaging ◽  
Lifetime Imaging ◽  
Confocal Fluorescence

Fluorescence axial nanotomography with plasmonics

2015 ◽  
Vol 178 ◽  
pp. 371-381 ◽  
Author(s):  
Nicholas I. Cade ◽  
Gilbert O. Fruhwirth ◽  
Alexey V. Krasavin ◽  
Tony Ng ◽  
David Richards
Keyword(s):  
Fluorescence Lifetime ◽  
Cell Imaging ◽  
Super Resolution ◽  
Cellular Membrane ◽  
Carcinoma Cells ◽  
Axial Position ◽  
Fluorescence Lifetime Imaging ◽  
Lifetime Imaging ◽  
Position Sensitivity ◽  
Confocal Fluorescence

We present a novel imaging technique with super-resolution axial sensitivity, exploiting the changes in fluorescence lifetime above a plasmonic substrate. Using conventional confocal fluorescence lifetime imaging, we show that it is possible to deliver down to 6 nm axial position sensitivity of fluorophores in whole biological cell imaging. We employ this technique to map the topography of the cellular membrane, and demonstrate its application in an investigation of receptor-mediated endocytosis in carcinoma cells.

scholarly journals A biosensor of local kinesin activity reveals roles of PKC and EB1 in KIF17 activation

2013 ◽  
Vol 203 (3) ◽  
pp. 445-455 ◽  
Author(s):  
Cedric Espenel ◽  
Bipul R. Acharya ◽  
Geri Kreitzer
Keyword(s):  
Energy Transfer ◽  
Fluorescence Lifetime ◽  
Single Cells ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Epithelial Differentiation ◽  
Fluorescence Lifetime Imaging ◽  
Lifetime Data ◽  
Lifetime Imaging ◽  
Phasor Plot

We showed previously that the kinesin-2 motor KIF17 regulates microtubule (MT) dynamics and organization to promote epithelial differentiation. How KIF17 activity is regulated during this process remains unclear. Several kinesins, including KIF17, adopt compact and extended conformations that reflect autoinhibited and active states, respectively. We designed biosensors of KIF17 to monitor its activity directly in single cells using fluorescence lifetime imaging to detect Förster resonance energy transfer. Lifetime data are mapped on a phasor plot, allowing us to resolve populations of active and inactive motors in individual cells. Using this biosensor, we demonstrate that PKC contributes to the activation of KIF17 and that this is required for KIF17 to stabilize MTs in epithelia. Furthermore, we show that EB1 recruits KIF17 to dynamic MTs, enabling its accumulation at MT ends and thus promoting MT stabilization at discrete cellular domains.

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