Abstract
The rapid development of scientific CMOS (sCMOS) technology has greatly advanced optical microscopy for biomedical research with superior sensitivity, resolution, field-of-view, and frame rates. However, for sCMOS sensors, the parallel charge-voltage conversion and different responsivity at each pixel induces extra readout and pattern noise compared to charge-coupled devices (CCD) and electron-multiplying CCD (EM-CCD) sensors. This can produce artifacts, deteriorate imaging capability, and hinder quantification of fluorescent signals, thereby compromising strategies to reduce photo-damage to live samples. Here, we propose a content-adaptive algorithm for the automatic correction of sCMOS-related noise (ACsN) for fluorescence microscopy. ACsN combines camera physics and layered sparse filtering to significantly reduce the most relevant noise sources in a sCMOS sensor while preserving the fine details of the signal. The method improves the camera performance, enabling fast, low-light and quantitative optical microscopy with video-rate denoising for a broad range of imaging conditions and modalities.
Highlights
The rapid development of scientific CMOS technology has greatly advanced optical microscopy for biomedical research with superior sensitivity, resolution, field-of-view, and frame rates
automatic correction of scientific complementary metal-oxide semiconductor (sCMOS)-related noise (ACsN) combines camera calibration, noise estimation and sparse filtering to correct the most relevant noise sources generated by a sCMOS camera (Fig. 1a and Supplementary Notes 1 and 2.1)
ACsN first corrects the fixed-pattern noise using a map of the offset and gain of the sCMOS pixels
Summary
The rapid development of scientific CMOS (sCMOS) technology has greatly advanced optical microscopy for biomedical research with superior sensitivity, resolution, field-of-view, and frame rates. 1234567890():,; The accurate acquisition of diverse anatomical and dynamic traits within a cell that span spatiotemporal scales provides insights into the fundamentals of living organisms In this context, scientific complementary metal-oxide semiconductor (sCMOS) cameras have rapidly been gaining popularity in optical microscopy for their higher frame rates, wider field-of-view, and lower electrical noise, compared to charge-coupled devices (CCD) or electron-multiplying CCDs (EM-CCD) cameras[1,2]. Scientific complementary metal-oxide semiconductor (sCMOS) cameras have rapidly been gaining popularity in optical microscopy for their higher frame rates, wider field-of-view, and lower electrical noise, compared to charge-coupled devices (CCD) or electron-multiplying CCDs (EM-CCD) cameras[1,2] Both CCD and CMOS cameras accumulate a signal charge in each pixel proportional to the local illumination intensity. These methods do not effectively remove the camera noise in many practical cases, either because of a tradeoff between noise correction and detail preservation[15] or the lack of a precise knowledge concerning the imaging system or the noise statistics[18,19]
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