Quantifying Molecular Dynamics within Complex Cellular Morphologies using LLSM-FRAP

Research output: Contribution to journalResearch articleContributedpeer-review


  • Huw Colin-York - , University of Oxford (Author)
  • John Heddleston - , Howard Hughes Medical Institute (Author)
  • Eric Wait - , Howard Hughes Medical Institute (Author)
  • Narain Karedla - , Rosalind Franklin Institute (Author)
  • Michael deSantis - , Howard Hughes Medical Institute (Author)
  • Satya Khuon - , Howard Hughes Medical Institute (Author)
  • Teng-Leong Chew - , Howard Hughes Medical Institute (Author)
  • Ivo F. Sbalzarini - , Chair of Scientific Computing for Systems Biology, Max Planck Institute of Molecular Cell Biology and Genetics (Author)
  • Marco Fritzsche - , University of Oxford, Rosalind Franklin Institute (Author)


Quantifying molecular dynamics within the context of complex cellular morphologies is essential toward understanding the inner workings and function of cells. Fluorescence recovery after photobleaching (FRAP) is one of the most broadly applied techniques to measure the reaction diffusion dynamics of molecules in living cells. FRAP measurements typically restrict themselves to single-plane image acquisition within a subcellular-sized region of interest due to the limited temporal resolution and undesirable photobleaching induced by 3D fluorescence confocal or widefield microscopy. Here, an experimental and computational pipeline combining lattice light sheet microscopy, FRAP, and numerical simulations, offering rapid and minimally invasive quantification of molecular dynamics with respect to 3D cell morphology is presented. Having the opportunity to accurately measure and interpret the dynamics of molecules in 3D with respect to cell morphology has the potential to reveal unprecedented insights into the function of living cells.


Original languageEnglish
Article number2200149
Number of pages11
JournalSmall methods
Issue number6
Publication statusPublished - 30 Jun 2022

External IDs

PubMed 35344286
ORCID /0000-0003-4414-4340/work/142252154