Superior performance with sCMOS over EMCCD in super-resolution optical fluctuation imaging

Area of Science:

• Optical microscopy
• Nanoscopy
• Biophysics

Background:

• Super-resolution optical fluctuation imaging (SOFI) extracts subdiffraction details from fluorescence intensity fluctuations.
• SOFI's reliance on fluctuation statistics presents unique detector requirements.
• Systematic evaluation of detectors for SOFI is lacking.

Purpose of the Study:

• To systematically evaluate the influence of detector parameters on SOFI performance.
• To compare scientific-grade complementary metal oxide semiconductor (sCMOS) cameras with other detectors for SOFI applications.
• To demonstrate the utility of sCMOS cameras for live-cell super-resolution imaging using SOFI.

Main Methods:

• Simulations and experimental tests were conducted to analyze detector influences.
• Key parameters investigated include pixel size, frame rate, noise levels, and gain.
• Performance was assessed based on spatiotemporal resolution and field-of-view.

Main Results:

• Smaller pixel size and faster readout speed of sCMOS cameras significantly improve SOFI's spatiotemporal resolution and field-of-view.
• SOFI performance is relatively insensitive to signal-to-noise ratio (SNR).
• The advantages of sCMOS in pixel size and readout speed outweigh SNR limitations compared to electron multiplying charge coupled devices (EMCCDs).

Conclusions:

• sCMOS cameras are superior to EMCCDs for SOFI, especially in live-cell super-resolution imaging.
• The large pixel count and fast frame rate of sCMOS enable high-quality, large field-of-view SOFI.
• This work provides crucial insights for selecting optimal detectors for advanced SOFI applications.