| Autor: |
Richardson DS; Department of NanoBiophotonics, Max Planck Institute for Biophysical Chemistry, 37077, Göttingen, Germany. drichardson@fas.harvard.edu.; Department of Molecular and Cellular Biology, Harvard Center for Biological Imaging, Harvard University, Cambridge, MA, 02138, USA. drichardson@fas.harvard.edu., Gregor C; Department of NanoBiophotonics, Max Planck Institute for Biophysical Chemistry, 37077, Göttingen, Germany., Winter FR; Department of NanoBiophotonics, Max Planck Institute for Biophysical Chemistry, 37077, Göttingen, Germany., Urban NT; Department of NanoBiophotonics, Max Planck Institute for Biophysical Chemistry, 37077, Göttingen, Germany., Sahl SJ; Department of NanoBiophotonics, Max Planck Institute for Biophysical Chemistry, 37077, Göttingen, Germany., Willig KI; Department of NanoBiophotonics, Max Planck Institute for Biophysical Chemistry, 37077, Göttingen, Germany.; Nanoscale Microscopy and Molecular Physiology of the Brain, University Medical Center Göttingen, 37075, Göttingen, Germany., Hell SW; Department of NanoBiophotonics, Max Planck Institute for Biophysical Chemistry, 37077, Göttingen, Germany. shell@gwdg.de.; Department of Optical Nanoscopy, Max Planck Institute for Medical Research, 69120, Heidelberg, Germany. shell@gwdg.de. |
| Abstrakt: |
Fluorescence-based biosensors have become essential tools for modern biology, allowing real-time monitoring of biological processes within living cells. Intracellular fluorescent pH probes comprise one of the most widely used families of biosensors in microscopy. One key application of pH probes has been to monitor the acidification of vesicles during endocytosis, an essential function that aids in cargo sorting and degradation. Prior to the development of super-resolution fluorescence microscopy (nanoscopy), investigation of endosomal dynamics in live cells remained difficult as these structures lie at or below the ~250 nm diffraction limit of light microscopy. Therefore, to aid in investigations of pH dynamics during endocytosis at the nanoscale, we have specifically designed a family of ratiometric endosomal pH probes for use in live-cell STED nanoscopy.Ratiometric fluorescent pH probes are useful tools to monitor acidification of vesicles during endocytosis, but the size of vesicles is below the diffraction limit. Here the authors develop a family of ratiometric pH sensors for use in STED super-resolution microscopy, and optimize their delivery to endosomes. |
