Taneja Chetna, George Jerin Geogy, Corsetti Stella, Wijesinghe Philip, Bruce Graham D, Zwart Maarten F, Bhattacharya Shanti, Dholakia Kishan
SUPA School of Physics and Astronomy, University of St Andrews, North Haugh, St Andrews, Fife KY16 9SS, UK.
Department of Electrical Engineering, IIT Madras, Chennai, India.
Biomed Opt Express. 2024 Oct 4;15(11):6183-6197. doi: 10.1364/BOE.538253. eCollection 2024 Nov 1.
The Bessel beam (BB) has found widespread adoption in various forms of light-sheet microscopy. However, for one-photon fluorescence, the transverse profile of the beam poses challenges due to the detrimental effect of the sidelobes. Here, we mitigate this issue by using a computer-generated phase element for generating a sidelobe suppressed Bessel beam (SSBB). We then progress to perform a comparison of biological imaging using SSBB to standard BB in a light-sheet geometry. The SSBB peak intensity is more than an order of magnitude higher than the first sidelobe. In contrast to a standard BB light-sheet, an SSBB does not need deconvolution. The SSBB propagates to depths exceeding 400 m in phantom samples maintaining a transverse size of 5 m. Finally, we demonstrate the advantage of using an SSBB light-sheet for biological applications by imaging fixed early-stage zebrafish larvae. In comparison to the standard BB, we observe a two-fold increase in contrast-to-noise ratio (CNR) when imaging the labelled cellular eye structures and the notochords. Our results provide an effective approach to generating and using SSBB light-sheets to enhance contrast for one-photon light-sheet microscopy.
贝塞尔光束（BB）已在各种形式的光片显微镜中得到广泛应用。然而，对于单光子荧光，由于旁瓣的有害影响，光束的横向轮廓带来了挑战。在这里，我们通过使用计算机生成的相位元件来生成旁瓣抑制贝塞尔光束（SSBB），从而缓解了这个问题。然后，我们在光片几何结构中，对使用SSBB进行生物成像与标准BB进行比较。SSBB的峰值强度比第一旁瓣高出一个数量级以上。与标准BB光片不同，SSBB不需要去卷积。SSBB在模拟样品中传播到超过400μm的深度，同时保持5μm的横向尺寸。最后，我们通过对固定的早期斑马鱼幼虫进行成像，展示了使用SSBB光片用于生物应用的优势。与标准BB相比，在对标记的细胞眼结构和脊索进行成像时，我们观察到对比度噪声比（CNR）提高了两倍。我们的结果提供了一种生成和使用SSBB光片以增强单光子光片显微镜对比度的有效方法。
