Single-molecule mountains yield nanoscale cell images

Abstract

Methods to simultaneously localize the positions of multiple single fluorophores by precisely determining their individual positions are now yielding impressive gains in fluorescence microscopy resolution.

Figure 1

Measuring mountain peaks for superresolution imaging. (a) Position-frequency image of single molecules of pentacene in a crystal, as a pumping laser beam is moved across the sample and the fluorescence is recorded. (b) It is easily possible to walk to the top of the cinder cone in Crater Lake to find the position of the peak accurately. (c) Similarly, in a living bacterial cell (blue), the image of a can determine its position (pink arrow) to much less than the width of the “mountain.” By turning single-molecule light sources like this on and off in a cell, researchers have recently obtained superresolution single fluorescent protein molecule appears as a wide spot due to fundamental diffraction effects, but one images. Scale bars: a, 5 × 10³ nm; b, ∼120 × 10⁹ nm; c, 250 nm.