Abstract
Temporal resolution is a key factor for imaging rapidly occurring events in biology. In this feature article, I investigate an approximate estimate for determining the temporal resolution limit. The condition that led to this limit is, the time taken by the ensemble (99.9%) of excited molecules to relax to ground state, assuming all the emitted photons are detected. In a simplistic three-level system, the temporal resolution is, ≈3τp, where τp = (loge10)/(kf + knr) and, kf and knr are respectively the radiative and non-radiative emission rates. This further assumes the ideal condition that, the quantum efficiency of the detector is unity and there are no other loses. We discuss few state-of-art microscopy techniques that are capable of high temporal resolution. This includes techniques such as multifocal multiphoton microscopy (MMM), multifocal plane microscopy, multiple excitation spot optical microscopy (MESO), multiplane microscopy and multiple light-sheet microscopy (MLSM).
Highlights
Fluorescence microscopy has emerged as one of the key imaging modalities in the last decade
Fluorescence imaging techniques that are capable of high temporal resolution are, multiphoton multifocal microscopy (MMM), multifocal plane microscopy, multiple excitation spot optical (MESO) microscopy and multiple light-sheet microscopy (MLSM)
Multifocal microscopy is probably the first fluorescence imaging technique that brings in the concept of high temporal resolution
Summary
Fluorescence microscopy has emerged as one of the key imaging modalities in the last decade. Of-late the field of super-resolution has expanded with the integration of super-resolution with light-sheet microscopy (Zanacchi et al, 2011; Deschout et al, 2014). Most of these techniques are capable of spatial super-resolution, some at the expense of poor temporal resolution. This is true for localization techniques (PALM, fPALM, STORM, GSDIM). Fluorescence imaging techniques that are capable of high temporal resolution are, multiphoton multifocal microscopy (MMM), multifocal plane microscopy, multiple excitation spot optical (MESO) microscopy and multiple light-sheet microscopy (MLSM).
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