TY - JOUR
T1 - Method to identify and minimize artifacts induced by fluorescent impurities in single-molecule localization microscopy
AU - Davis, Janel L.
AU - Dong, Biqin
AU - Sun, Cheng
AU - Zhang, Hao F.
N1 - Funding Information:
This work was supported by NSF Grant Nos. CBET-1706642, EFRI-1830969, and EEC-1530734 and Northwestern University Innovative Initiative Incubator (I3) Award. JLD was supported by the NSF Graduate Research Fellowship 1000231682 and NIH predoctoral Biotechnology Training Grant No. T32GM008449. Plasma cleaned and UV-Ozone cleaned coverslips were prepared in the EPIC facility of Northwestern University’s NUANCE Center, which received support from the Soft and Hybrid Nanotechnology Experimental (SHyNE) Resource (NSF ECCS-1542205), the MRSEC program (NSF DMR-1121262) at the Materials Research Center, the International Institute for Nanotechnology (IIN), the Keck Foundation, and the State of Illinois.
Publisher Copyright:
© 2018 The Authors.
PY - 2018/10/1
Y1 - 2018/10/1
N2 - The existence of fluorescent impurities has been a long-standing obstacle in single-molecule imaging, which results in sample misidentification and higher localization uncertainty. Spectroscopic single-molecule localization microscopy can record the full fluorescent spectrum of every stochastic single-molecule emission event. This capability allows us to quantify the spatial and spectral characteristics of fluorescent impurities introduced by sample preparation steps, based on which we developed a method to effectively separate fluorescent impurities from target molecules.
AB - The existence of fluorescent impurities has been a long-standing obstacle in single-molecule imaging, which results in sample misidentification and higher localization uncertainty. Spectroscopic single-molecule localization microscopy can record the full fluorescent spectrum of every stochastic single-molecule emission event. This capability allows us to quantify the spatial and spectral characteristics of fluorescent impurities introduced by sample preparation steps, based on which we developed a method to effectively separate fluorescent impurities from target molecules.
KW - fluorescence spectral imaging
KW - fluorescent impurities
KW - single-molecule localization microscopy
KW - single-molecule spectroscopy
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U2 - 10.1117/1.JBO.23.10.106501
DO - 10.1117/1.JBO.23.10.106501
M3 - Article
C2 - 30334394
AN - SCOPUS:85055071573
SN - 1083-3668
VL - 23
JO - Journal of Biomedical Optics
JF - Journal of Biomedical Optics
IS - 10
M1 - 106501
ER -