提出新算法,让低光下活细胞成像更清晰更快。
Hybrid iterating-averaging low photon budget Gabor holographic microscopy
- 用迭代平均法降低低光照下的噪声和孪生像干扰
- 实验验证可清晰拍摄高速游动的精子细胞
- 适合弱光、薄样本成像,对生物医学研究很实用
活细胞成像面临高对比度与低扰动的双重挑战。定量相位成像(QPI)技术可在无标记情况下实现高对比成像,而低光子预算(LPB)成像可进一步减少光刺激、光毒性及光损伤,支持高速成像。然而,在低光条件下,相机散粒噪声与全息孪生像干扰问题严重制约了数字白光全息显微镜(DIHM)的性能。本文提出一种创新的迭代吉布斯平均(IGA)算法,专为多帧低光条件下的DIHM设计。仿真结果显示,IGA在高噪声环境下显著优于传统算法。实验验证中,该算法成功实现了在极低光照下对动态精子细胞及相位测试靶标的高速成像,并有效重建了光学厚度极小的样本,这些样本即使在高光子预算下也常因信噪比过低难以成像。本方法推动了无光刺激、高速生物样本成像的发展,拓展了生物医学与环境成像的应用边界。
原文摘要 · Abstract (English)
One of the primary challenges in live cell culture observation is achieving high-contrast imaging with minimal impact on sample behavior. Quantitative phase imaging (QPI) techniques address this by providing label-free, high-contrast images of transparent samples. Measurement system influence may be further reduced by imaging samples under low illumination intensity (low photon budget - LPB), thereby minimizing photostimulation, phototoxicity, and photodamage, and enabling high-speed imaging. LPB imaging is challenging in QPI due to significant camera shot noise and quantification noise. Digital in-line holographic microscopy (DIHM), working with or without lenses, is a QPI technique known for its robustness to quantification noise. However, reducing simultaneously the camera shot noise and the inherent in-line holographic twin image disturbances remain a critical, yet unaddressed, challenge. This study introduces an innovative iterative Gabor averaging (IGA) algorithm designed specifically for filling this important scientific gap in multi-frame DIHM under LPB conditions. We evaluated the performance of the IGA on simulated data showing that it outperformed traditional algorithms in terms of reconstruction accuracy under high noise conditions. Those results were corroborated by experimental validation involving high-speed imaging of dynamic sperm cells and a phase test target under significantly reduced illumination power. Additionally, the IGA algorithm proved successful in reconstructing optically thin samples, which typically produce low signal-to-noise ratio holograms even under high photon budget conditions. These advancements facilitate photostimulation-free and high-speed imaging of dynamic biological samples and enhance the capability to image samples with extremely low optical thickness, potentially transforming various applications in biomedical and environmental imaging.
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