Reversible Cryo-arrests of Living Cells to Pause Molecular Movements for High-resolution Imaging

Reversible Cryo-arrests of Living Cells to Pause Molecular Movements for High-resolution Imaging

Blog Article

Fluorescence live-cell imaging by single molecule localization microscopy (SMLM) or fluorescence lifetime imaging microscopy (FLIM) in principle allows for the spatio-temporal observation of molecular patterns in individual, living cells.However, the dynamics of mel axolotl molecules within cells hamper their precise observation.We present here a detailed protocol for consecutive cycles of reversible cryo-arrest of apunisw2 living cells on a microscope that allows for a precise determination of the evolution of molecular patterns within individual living cells.The usefulness of this approach has been demonstrated by observing ligand-induced clustering of receptor tyrosine kinases as well as their activity patterns by SMLM and FLIM (Masip et al.

, 2016).