# Advanced fluorescence lifetime-enhanced multiplexed nanoscopy of cells

## TL;DR

This paper reviews recent progress in fluorescence lifetime imaging, highlighting its use in high-resolution cell imaging and multiplexed super-resolution techniques.

## Contribution

The paper introduces advancements in wide-field FLIM using the LINCam system for single-molecule sensitivity and super-resolution imaging.

## Key findings

- Single-molecule FLIM in the far-red spectral region enables detailed cellular imaging.
- Fast and multiplexed super-resolution imaging of cells is now achievable with improved precision.
- Three-dimensional imaging with high axial localization precision has been enhanced using new detector technologies.

## Abstract

In this review paper, we summarize the significant advancements in the field of fluorescence lifetime imaging microscopy (FLIM), particularly wide-field FLIM with single-molecule sensitivity, achieved using the time-correlated single-photon counting-based position-sensitive LINCam system. Fluorescence lifetime adds valuable information beyond conventional intensity-based imaging, enabling diverse applications across research fields. Here, we focus on three primary bioimaging applications: (I) single-molecule FLIM in the far-red spectral region, (II) fast and multiplexed super-resolution imaging of cells, and (III) three-dimensional super-resolution imaging with high axial localization precision. Recent advances in position-sensitive detector technologies offer exciting opportunities for high-throughput super-resolution imaging with enhanced localization precision.