High contrast, isotropic, and uniform 3D-imaging of centimeter-scale scattering samples using structured illumination light-sheet microscopy with axial sweeping

被引:4
|
作者
Frantz, David [1 ]
Karamahmutoglu, Tugba [2 ]
Schaser, Allison J. [3 ]
Kirik, Deniz [2 ]
Berrocal, Edouard [1 ]
机构
[1] Lund Univ, Dept Phys, Div Combust Phys, Lund, Sweden
[2] Lund Univ, Dept Expt Med Sci, Brain Repair & Imaging Neural Syst BRAINS Unit, BMC D11, S-22184 Lund, Sweden
[3] Purdue Univ, Purdue Inst Integrat Neurosci, Dept Speech Language & Hearing Sci, 207 S Martin Jischke Dr DLR 335, W Lafayette, IN 47907 USA
来源
BIOMEDICAL OPTICS EXPRESS | 2022年 / 13卷 / 09期
基金
美国国家卫生研究院;
关键词
LASER ILLUMINATION; PLANE ILLUMINATION; TURBID MEDIA; RECONSTRUCTION; SUPPRESSION;
D O I
10.1364/BOE.464039
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Light-sheet fluorescent microscopy (LSFM) has, in recent years, allowed for rapid 3D-imaging of cleared biomedical samples at larger and larger scale. However, even in cleared samples, multiple light scattering often degrades the imaging contrast and widens the optical sectioning. Accumulation of scattering intensifies these negative effects as light propagates inside the tissue, which accentuates the issues when imaging large samples. With axially swept light-sheet microscopy (ASLM), centimeter-scale samples can be scanned with a uniform micrometric optical sectioning. But to fully utilize these benefits for 3D-imaging in biomedical tissue samples, suppression of scattered light is needed. Here, we address this by merging ASLM with light-sheet based structured illumination into Structured Illumination Light-sheet Microscopy with Axial Sweeping (SILMAS). The SILMAS method thus enables high-contrast imaging, isotropic micrometric resolution and uniform optical sectioning in centimeter-scale scattering samples, creating isotropic 3D-volumes of e.g., whole mouse brains without the need for any computation-heavy post-processing. We demonstrate the effectiveness of the approach in agarose gel phantoms with fluorescent beads, and in an PFF injected alpha-synuclein transgenic mouse model tagged with a green fluorescent protein (SynGFP). SILMAS imaging is compared to standard ASLM imaging on the same samples and using the same optical setup, and is shown to increase contrast by as much as 370% and reduce widening of optical sectioning by 74%. With these results, we show that SILMAS improves upon the performance of current state-of-the-art light-sheet microscopes for large and imperfectly cleared tissue samples and is a valuable addition to the LSFM family.
引用
收藏
页码:4907 / 4925
页数:19
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