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.
Sidelobe suppressed Bessel beams for one-photon light-sheet microscopy.
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作者:Taneja Chetna, George Jerin Geogy, Corsetti Stella, Wijesinghe Philip, Bruce Graham D, Zwart Maarten F, Bhattacharya Shanti, Dholakia Kishan
| 期刊: | Biomedical Optics Express | 影响因子: | 3.200 |
| 时间: | 2024 | 起止号: | 2024 Oct 4; 15(11):6183-6197 |
| doi: | 10.1364/BOE.538253 | ||
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