Adaptive optics in an oblique plane microscope

IF 2.9 2区 医学 Q2 BIOCHEMICAL RESEARCH METHODS Biomedical optics express Pub Date : 2024-06-28 DOI:10.1364/boe.524013
Conor Mcfadden, Zach Marin, Bingying Chen, Stephan Daetwyler, Xiaoding Wang, Divya Rajendran, Kevin M. Dean, Reto Fiolka
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Abstract

Adaptive optics (AO) can restore diffraction-limited performance when imaging beyond superficial cell layers in vivo and in vitro, and as such, is of interest for advanced 3D microscopy methods such as light-sheet fluorescence microscopy (LSFM). In a typical LSFM system, the illumination and detection paths are separate and subject to different optical aberrations. To achieve optimal microscope performance, it is necessary to sense and correct these aberrations in both light paths, resulting in a complex microscope system. Here, we show that in an oblique plane microscope (OPM), a type of LSFM with a single primary objective lens, the same deformable mirror can correct both illumination and fluorescence detection. Besides reducing the complexity, we show that AO in OPM also restores the relative alignment of the light-sheet and focal plane, and that a projection imaging mode can stabilize and improve the wavefront correction in a sensorless AO format. We demonstrate OPM with AO on fluorescent nanospheres and by imaging the vasculature and cancer cells in zebrafish embryos embedded in a glass capillary, restoring diffraction limited resolution and improving the signal strength twofold.
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斜面显微镜中的自适应光学技术
自适应光学(AO)可以恢复体内和体外浅表层细胞成像时的衍射限制性能,因此对光片荧光显微镜(LSFM)等先进的三维显微方法很有意义。在典型的 LSFM 系统中,照明和检测路径是分开的,并受到不同光学像差的影响。为了获得最佳的显微镜性能,有必要感测和校正两条光路中的这些像差,从而形成一个复杂的显微镜系统。在这里,我们展示了在斜面显微镜(OPM)(一种具有单主物镜的 LSFM)中,同一可变形镜可以同时校正照明和荧光检测。除了降低复杂性外,我们还展示了 OPM 中的自动光学还能恢复光片和焦平面的相对对齐,而且投影成像模式可以稳定和改进无传感器自动光学格式中的波前校正。我们演示了在荧光纳米球上使用 AO 的 OPM,并通过对嵌入玻璃毛细管中的斑马鱼胚胎的血管和癌细胞进行成像,恢复了衍射受限的分辨率,并将信号强度提高了两倍。
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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.
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