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Related Experiment Video

Updated: Apr 24, 2026

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Effects of fixed pattern noise on single molecule localization microscopy.

F Long1, S Q Zeng, Z L Huang

  • 1Britton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics-Huazhong University of Science and Technology, Wuhan 430074, China. leo@mail.hust.edu.cn.

Physical Chemistry Chemical Physics : PCCP
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Summary

Scientific complementary metal oxide semiconductor (sCMOS) cameras offer fast imaging for single molecule localization microscopy. Fixed pattern noise (FPN) minimally impacts localization precision but introduces negligible bias (<2 nm) in most applications.

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Area of Science:

  • Biophysics
  • Optical Microscopy
  • Super-resolution Imaging

Background:

  • Scientific complementary metal oxide semiconductor (sCMOS) cameras enable faster imaging in single molecule localization microscopy (SMLM) compared to charge-coupled device (CCD) cameras.
  • Fixed pattern noise (FPN) in sCMOS cameras can compromise image uniformity, potentially hindering their widespread adoption in SMLM.

Purpose of the Study:

  • To quantitatively assess the impact of FPN on the performance of sCMOS cameras in SMLM.
  • To evaluate the effects of FPN on localization precision and localization bias in SMLM.

Main Methods:

  • Simulations and experimental measurements were used to investigate FPN effects.
  • Localization precision and bias were analyzed under varying FPN conditions.

Main Results:

  • FPN was found to have a negligible effect on localization precision in SMLM.
  • FPN introduces a small localization bias, typically less than 2 nm for a commercial sCMOS camera.

Conclusions:

  • The localization bias introduced by FPN in sCMOS cameras is generally negligible for most SMLM applications.
  • This study alleviates concerns about FPN, promoting the use of sCMOS cameras in SMLM.