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Localization-based super-resolution microscopy with an sCMOS camera part II: experimental methodology for comparing
Fan Long1, Shaoqun Zeng, Zhen-Li Huang
1Britton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics-Huazhong University of Science and Technology, Wuhan 430074, China.
Optics Express
|October 6, 2012
Summary
Scientific CMOS cameras show superior performance for super-resolution microscopy compared to EMCCD cameras. This study provides a methodology to quantitatively compare low-light camera performance in single molecule detection and localization.
Area of Science:
- Optical Microscopy
- Biophysics
- Imaging Technology
Background:
- Localization-based super-resolution microscopy demands high-performance low-light cameras.
- Debate exists on whether scientific CMOS (sCMOS) cameras are suitable for this application.
- Electron Multiplying Charge Coupled Device (EMCCD) cameras are a popular choice.
Purpose of the Study:
- To develop and apply a quantitative methodology for comparing low-light camera performance.
- To evaluate the suitability of sCMOS cameras for localization-based super-resolution microscopy.
- To compare sCMOS and EMCCD camera performance in single molecule detection and localization.
Main Methods:
- Established an experimental methodology for quantitative camera performance comparison.
- Characterized single molecule detection using image signal-to-noise ratio (SNR).
- Assessed localization performance via localization accuracy.
Main Results:
- A newly launched sCMOS camera demonstrated superior imaging performance.
- The sCMOS camera outperformed a popular EMCCD camera.
- Superior performance was observed within a signal range (15-12000 photons/pixel) relevant for super-resolution microscopy.
Conclusions:
- Scientific CMOS cameras offer advantages for localization-based super-resolution microscopy.
- The developed methodology provides a reliable framework for camera evaluation.
- sCMOS cameras are a viable and potentially superior alternative to EMCCD cameras for this technique.
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