FL!P™ microscopy allows observations of changes in protein conformation, in living cells, without the need to modify the protein of interest. The technique relies on FL!Ps™: molecular biosensors designed specifically for polarization-resolved fluorescence microscopy. The biosensors are complemented by a suitable implementation of polarization-resolved fluorescence microscopy. The breakthrough results that paved the way to FL!P development are described in detail in Lazar J., Bondar A., Timr S., and Firestein S.J., Nature Methods (2011).
Briefly, light absorption by most molecules is anisotropic. So, if an assembly of molecules is not oriented completely randomly, it will absorb light of different polarizations to different extents. Absorption of different linear polarizations to different extents is called 'linear dichroism'. An assembly of molecules that is not oriented completely randomly will also emit fluorescence that is polarized. Both linear dichroism and fluorescence polarization can be used for observing FL!P™ biosensors (that is, for performing FL!P™ microscopy).
We provide everything you need for FL!P™ microscopy: the FL!P™ biosensors, the microscopy equipment (the 1P FL!Pper, the 2P FL!Pper, and more coming soon), as well as the software (Polaris/Polaris+).
Some publications describing FL!Ps and polarization microscopy:
Lazar J., Bondar A., Timr S., Firestein S.J.:
Two-photon polarization microscopy reveals protein structure and function.
Nature Methods 8: 684-690, 2011.
Timr S., Bondar A., Cwiklik L., Stefl M., Hof M., Vazdar M., Lazar J., Jungwirth P.:
Accurate Determination of the Orientational Distribution of a Fluorescent Molecule in a Phospholipid Membrane.
Journal of Physical Chemistry B 118: 855-863, 2013.
Bondar A., Lazar J.:
Dissociated G GTP and G Protein Subunits Are the Major Activated Form of Heterotrimeric Gi/o Proteins.
Journal of Biological Chemistry 289: 1271-1281, 2014.
Han Z., Jin L., Chen F, Loturco J.J., Cohen L.B., Bondar A., Lazar J., Pieribone V.:
Mechanistic Studies of the Genetically Encoded Fluorescent Protein Voltage Probe ArcLight.
PLoS One 9: e113873, 2014.
Bondar A, Rybakova O, Melcr J, Dohnálek J, Khoroshyy P, Ticháček O, Timr Š, Miclea P, Sakhi A, Marková V, Lazar J.:
Quantitative linear dichroism imaging of molecular processes in living cells made simple by open software tools.
Communications Biology: 2021 Feb 12;4(1):189.
Briefly, light absorption by most molecules is anisotropic. So, if an assembly of molecules is not oriented completely randomly, it will absorb light of different polarizations to different extents. Absorption of different linear polarizations to different extents is called 'linear dichroism'. An assembly of molecules that is not oriented completely randomly will also emit fluorescence that is polarized. Both linear dichroism and fluorescence polarization can be used for observing FL!P™ biosensors (that is, for performing FL!P™ microscopy).
We provide everything you need for FL!P™ microscopy: the FL!P™ biosensors, the microscopy equipment (the 1P FL!Pper, the 2P FL!Pper, and more coming soon), as well as the software (Polaris/Polaris+).
Some publications describing FL!Ps and polarization microscopy:
Lazar J., Bondar A., Timr S., Firestein S.J.:
Two-photon polarization microscopy reveals protein structure and function.
Nature Methods 8: 684-690, 2011.
Timr S., Bondar A., Cwiklik L., Stefl M., Hof M., Vazdar M., Lazar J., Jungwirth P.:
Accurate Determination of the Orientational Distribution of a Fluorescent Molecule in a Phospholipid Membrane.
Journal of Physical Chemistry B 118: 855-863, 2013.
Bondar A., Lazar J.:
Dissociated G GTP and G Protein Subunits Are the Major Activated Form of Heterotrimeric Gi/o Proteins.
Journal of Biological Chemistry 289: 1271-1281, 2014.
Han Z., Jin L., Chen F, Loturco J.J., Cohen L.B., Bondar A., Lazar J., Pieribone V.:
Mechanistic Studies of the Genetically Encoded Fluorescent Protein Voltage Probe ArcLight.
PLoS One 9: e113873, 2014.
Bondar A, Rybakova O, Melcr J, Dohnálek J, Khoroshyy P, Ticháček O, Timr Š, Miclea P, Sakhi A, Marková V, Lazar J.:
Quantitative linear dichroism imaging of molecular processes in living cells made simple by open software tools.
Communications Biology: 2021 Feb 12;4(1):189.