用于单光子光片显微镜的侧射抑制贝塞尔光束。

IF 2.9 2区 医学 Q2 BIOCHEMICAL RESEARCH METHODS Biomedical optics express Pub Date : 2024-10-04 eCollection Date: 2024-11-01 DOI:10.1364/BOE.538253
Chetna Taneja, Jerin Geogy George, Stella Corsetti, Philip Wijesinghe, Graham D Bruce, Maarten F Zwart, Shanti Bhattacharya, Kishan Dholakia
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引用次数: 0

摘要

贝塞尔光束(BB)已被广泛应用于各种形式的光片显微镜。然而,对于单光子荧光来说,光束的横向剖面会因侧叶的不利影响而带来挑战。在这里,我们通过使用计算机生成的相位元件来生成侧叶抑制贝塞尔光束(SSBB),从而缓解了这一问题。然后,我们将使用 SSBB 与光片几何中的标准 BB 进行生物成像比较。SSBB 的峰值强度比第一个侧叶高出一个数量级以上。与标准 BB 光片相比,SSBB 不需要解卷积。在保持横向尺寸为 5 μm 的幻影样本中,SSBB 的传播深度超过 400 μm。最后,我们通过对固定的早期斑马鱼幼体进行成像,证明了在生物应用中使用 SSBB 光片的优势。与标准 BB 相比,我们观察到在对标记的细胞眼结构和notochords 进行成像时,对比度-噪声比(CNR)提高了两倍。我们的研究结果为生成和使用 SSBB 光片来增强单光子光片显微镜的对比度提供了一种有效的方法。
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Sidelobe suppressed Bessel beams for one-photon light-sheet microscopy.

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 μm in phantom samples maintaining a transverse size of 5 μm. 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.

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来源期刊
Biomedical optics express
Biomedical optics express BIOCHEMICAL RESEARCH METHODS-OPTICS
CiteScore
6.80
自引率
11.80%
发文量
633
审稿时长
1 months
期刊介绍: The journal''s scope encompasses fundamental research, technology development, biomedical studies and clinical applications. BOEx focuses on the leading edge topics in the field, including: Tissue optics and spectroscopy Novel microscopies Optical coherence tomography Diffuse and fluorescence tomography Photoacoustic and multimodal imaging Molecular imaging and therapies Nanophotonic biosensing Optical biophysics/photobiology Microfluidic optical devices Vision research.
期刊最新文献
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