TY - JOUR
T1 - Fluorescence Lifetime Multiplex Imaging in Expansion Microscopy with Tunable Donor-Acceptor Polymer Dots
AU - Liu, Jie
AU - Liu, Zhihe
AU - Mi, Feixue
AU - Yao, Zihan
AU - Fang, Xiaofeng
AU - Wang, Yingjie
AU - Zhao, Zhongying
AU - Wu, Changfeng
N1 - Funding information:
This work was financially supported by the National Key R&D Program of China (Grant No. 2020YFA0909000), the National Natural Science Foundation of China (Grant No. 62235007, 22204070), the Shenzhen Science and Technology Program (Grant No. KQTD20170810111314625, JCYJ20210324115807021, SGDX20211123114002003), the Shenzhen Bay Laboratory (SZBL2021080601002), the Guangdong-Hong Kong Technology Cooperation Funding Scheme (Grant No. GHP/176/21SZ) and the Guangdong Provincial Key Laboratory of Advanced Biomaterials (2022B1212010003). The authors acknowledge the assistance of SUSTech Core Research Facilities.
Publisher Copyright:
© 2023 The Authors. Co-published by Nanjing University and American Chemical Society.
PY - 2023/9/25
Y1 - 2023/9/25
N2 - Fluorescence lifetime imaging microscopy (FLIM) has been widely used in cell biology to detect biomolecules and their interactions. However, breaking the diffraction limit remains a challenge in FLIM due to the typically required photon counting method and the limited photon output of conventional dyes. Here, we introduce semiconducting polymer dots (Pdots) for fluorescence lifetime imaging in expansion microscopy by virtue of their tunable lifetime and huge photon budget. We developed three fluorescent Pdots with average lifetimes ranging from 0.4 to 5 ns by varying the polymer species and compositions. Despite their large spectral overlap, distinctive distributions of the Pdots can be resolved in the lifetime domain. The high fluorescence brightness and large photon output offered by Pdots enable multiplex lifetime imaging in photon-starved expansion microscopy, by which subcellular structures were resolved with a spatial resolution of ∼49 nm. This study reveals the potential of the tunable Pdot probes for lifetime multiplex imaging in expansion microscopy.
AB - Fluorescence lifetime imaging microscopy (FLIM) has been widely used in cell biology to detect biomolecules and their interactions. However, breaking the diffraction limit remains a challenge in FLIM due to the typically required photon counting method and the limited photon output of conventional dyes. Here, we introduce semiconducting polymer dots (Pdots) for fluorescence lifetime imaging in expansion microscopy by virtue of their tunable lifetime and huge photon budget. We developed three fluorescent Pdots with average lifetimes ranging from 0.4 to 5 ns by varying the polymer species and compositions. Despite their large spectral overlap, distinctive distributions of the Pdots can be resolved in the lifetime domain. The high fluorescence brightness and large photon output offered by Pdots enable multiplex lifetime imaging in photon-starved expansion microscopy, by which subcellular structures were resolved with a spatial resolution of ∼49 nm. This study reveals the potential of the tunable Pdot probes for lifetime multiplex imaging in expansion microscopy.
KW - expansion microscopy
KW - fluorescence lifetime imaging microscopy
KW - fluorescence probes
KW - semiconductor polymer dots
KW - super-resolution microscopy
UR - http://www.scopus.com/inward/record.url?scp=85188732069&partnerID=8YFLogxK
U2 - 10.1021/cbmi.3c00058
DO - 10.1021/cbmi.3c00058
M3 - Journal article
AN - SCOPUS:85188732069
SN - 2832-3637
VL - 1
SP - 550
EP - 557
JO - Chemical and Biomedical Imaging
JF - Chemical and Biomedical Imaging
IS - 6
ER -