Förster resonance energy transfer (FRET) and fluorescence lifetime imaging (FLIM) have been coupled with multiphoton microscopy to image in vivo dynamics. However, the increase in optical aberrations as a function of depth significantly reduces the fluorescent signal, spatial resolution, and fluorescence lifetime accuracy. We present the development of a time-resolved FRET-FLIM imaging system with adaptive optics. We demonstrate the improvement of our adaptive optics (AO)-FRET-FLIM instrument over standard multiphoton FRET-FLIM imaging. We validate our approach using fixed cellular samples with FRET standards and in vivo with live imaging in a mouse kidney.
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| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7340371 | PMC |
| http://dx.doi.org/10.1364/OL.385950 | DOI Listing |
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