意大利热那亚技术研究所Giuseppe Vicidomini团队近日揭示了阵列检测可以实现简单而强大的MINFLUX的大定位范围。这一研究成果于2025年7月3日发表在《光：科学与应用》杂志上。

MINFLUX概念通过克服荧光团光子计数的限制，显著提高了单分子定位显微镜（SMLM）的定位性能。典型的MINFLUX显微镜通过以圆形轨迹扫描分子周围的零强度焦点来定位目标分子，较小的轨迹直径会在给定数量的光子下产生更好的定位不确定性。由于这种方法要求分子在扫描轨迹内，MINFLUX通常依赖于轨迹直径减小的迭代方案。 这种迭代方法容易导致轨迹错位，增加了系统的复杂性。

研究组介绍了ISM-FLUX，这是一种使用图像扫描显微镜（ISM）和单光子雪崩二极管阵列探测器的MINFLUX的新实现。ISM-FLUX在轨迹内提供精确的MINFLUX定位，同时在轨迹外保持传统的光子限制不确定性。ISM-FLUX定位的鲁棒性导致更大的定位范围，并大大简化了架构，这可能有助于更广泛地采用MINFLUX。

附：英文原文

Title: Array detection enables large localization range for simple and robust MINFLUX

Author: Slenders, Eli, Patil, Sanket, Held, Marcus Oliver, Zunino, Alessandro, Vicidomini, Giuseppe

Issue&Volume: 2025-07-03

Abstract: The MINFLUX concept significantly improves the localization properties of single-molecule localization microscopy (SMLM) by overcoming the limit imposed by the fluorophore’s photon counts. Typical MINFLUX microscopes localize the target molecule by scanning a zero-intensity focus around the molecule in a circular trajectory, with smaller trajectory diameters yielding better localization uncertainties for a given number of photons. Since this approach requires the molecule to be within the scanned trajectory, MINFLUX typically relies on an iterative scheme with decreasing trajectory diameters. This iterative approach is prone to misplacements of the trajectory and increases the system’s complexity. In this work, we introduce ISM-FLUX, a novel implementation of MINFLUX using image-scanning microscopy (ISM) with a single-photon avalanche diode array detector. ISM-FLUX provides a precise MINFLUX localization within the trajectory while maintaining a conventional photon-limited uncertainty outside it. The robustness of ISM-FLUX localization results in a larger localization range and greatly simplifies the architecture, which may facilitate broader adoption of MINFLUX.

DOI: 10.1038/s41377-025-01883-1

Source: https://www.nature.com/articles/s41377-025-01883-1