Toward an Axial Nanoscale Ruler for Fluorescence Microscopy

• Published in: ACS nano Vol. 11; no. 12; pp. 11762 - 11767
• Main Authors: Simoncelli, Sabrina, Makarova, Maria, Wardley, William, Owen, Dylan M
• Format: Journal Article
• Language: English
• Published: United States American Chemical Society 26.12.2017
• ISSN: 1936-0851; 1936-086X; 1936-086X
• DOI: 10.1021/acsnano.7b07133

In the discussion of resolution in optical microscopy, axial precision has often come second to its lateral counterpart. However, biological systems make no special arrangements for our preferred direction of imaging. The ability to measure axial distances, that is, the heights of fluorophores relative to a plane of reference, is thus of paramount importance and has been the subject of several recent advances. A novel method is to modify the fluorescence emission based on the height of the individual fluorophore, such that its z-position is encoded somehow in the detected signal. One such approach is metal-enhanced energy transfer, recently extended to multicolor distance measurements and applied to study the topography of the nuclear membrane. Here, the fluorescence lifetime is shortened due to the proximity of the fluorophores to a thin metallic surface. Fluorescence lifetime imaging can therefore be used as an axial ruler with nanometer precision.