Rapid Depolarization‐Free Nanoscopic Background Elimination of Cellular Metallic Nanoprobes

Abstract

The crowded intracellular environment of biomolecules, including organelles, solutes, proteins, and membranes, presents distinct biomolecular dynamics crucial for the functions of biomolecules within living cells. However, background suppression is critical to uncover nanoscale dynamics in living cells. Herein, a new method for enabling rapid, nanoscale background elimination of cellular metallic nanoprobes is presented. By employing integrated nanoscopic correction (iNC) designed to eliminate depolarization effects, which compromise background elimination, a real‐time algorithm to subtract and increment orthogonal pairs of polarizations for real‐time nanoscale background elimination is introduced. The ability to analyze orthogonal pairs at high speed over the entire polarization range is currently difficult to achieve using conventional methods. By processing orthogonal pairs in real time, this method minimizes movement artifacts during the background elimination process. Nanometer spatial stability which enables two orders of magnitude increase in signal‐to‐noise ratio of cellular metallic nanoprobes is shown. Nanoscale background elimination aiding the ability to accurately track biomolecules and their dynamics in living cells is anticipated.

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