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Effects of fixed pattern noise on single molecule localization microscopy
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
|September 6, 2014
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.
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.
