# High-Throughput Hyperspectral and Multiplexed Super-Resolution Fluorescence Imaging by SP-STORM

## TL;DR

A new microscopy method called SP-STORM enables fast, high-resolution imaging of multiple subcellular structures simultaneously.

## Contribution

SP-STORM introduces a novel approach combining spectral and spatial information for rapid, high-throughput super-resolution imaging.

## Key findings

- SP-STORM simultaneously images five subcellular structures with minimal crosstalk.
- The method is over ten times faster than existing multiplexing microscopy techniques.
- SP-STORM is compatible with other super-resolution microscopy approaches.

## Abstract

Simultaneous determination of spatial location and spectral
color
of single molecules at large molecular density with high throughput
was achieved by combining single-molecule photoswitching and optical
in-hardware Fourier transformation of single-molecule emission spectra
into the phasor space. The method, named as spectral phasor enabled
stochastic optical reconstruction microscopy (SP-STORM), achieved
simultaneous super-resolution imaging of five subcellular structures
in parallel with minimum crosstalk for the first time. The high-throughput
feature of SP-STORM enables these subcellular structures to be readily
resolved in about one min, which is more than an order of magnitude
faster than other multiplexing single-molecule localization microscopy
techniques. The concept of SP-STORM is also compatible with and can
be readily applicable to other super-resolution microscopy.