A quantitative analysis of various patterns applied in lattice light sheet microscopy.
A quantitative analysis of various patterns applied in lattice light sheet microscopy.
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DOI：
10.1038/s41467-022-32341-w
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发表时间：
2022-08-08
影响因子：
16.6
通讯作者：
中科院分区：
文献类型：
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与宽场和共焦显微镜相比，光片显微镜可降低光毒性和背景，并提高成像速度。然而，当配备高斯光束时，光片显微镜的轴向分辨率和可观察视野成反比。与高斯光束相比，基于抖动光学晶格的光片通过使用轴向结构化照明图案提高了轴向分辨率和光束均匀性。然而，这些优点是以增加对样本的总照明和减少照明图案的轴向限制为代价的。通过在固定细胞和活细胞中进行模拟和实验测量，我们量化了高斯光片和晶格光片在光束均匀性、轴向分辨率、横向分辨率和光漂白方面的差异。我们演示了如何调整不同的光学晶格照明模式以优先考虑轴向分辨率或光学切片。最后，我们介绍了一种对具有互补光学特性的不同晶格光片图案进行光谱融合的连续采集的方法，以实现高分辨率和低背景图像。与宽场和共焦显微镜相比，光片显微镜可降低光毒性和背景，同时提高成像速度。在这里，作者利用固定细胞和活细胞中的模拟和实验数据量化了高斯光片和晶格光片之间的差异。
Light sheet microscopes reduce phototoxicity and background and improve imaging speed compared to widefield and confocal microscopes. However, when equipped with Gaussian beams, the axial resolving power of a light sheet microscope and the observable field of view are inversely related. Light sheets based on dithered optical lattices improve axial resolution and beam uniformity compared Gaussian beams by using axially structured illumination patterns. However, these advantages come at the expense of an increased total illumination to the specimen and a decreased axial confinement of the illumination pattern. Using simulations and experimental measurements in fixed and live cells, we quantify the differences between Gaussian and lattice light sheets on beam uniformity, axial resolution, lateral resolution, and photobleaching. We demonstrate how different optical lattice illumination patterns can be tuned to prioritize either axial resolution or optical sectioning. Finally, we introduce an approach to spectrally fuse sequential acquisitions of different lattice light sheet patterns with complementary optical properties to achieve both high resolution and low background images. Light sheet microscopes reduce phototoxicity and background while improving imaging speed compared to widefield and confocal microscopes. Here the authors quantify the differences between Gaussian and lattice light sheets using simulations and experimental data in fixed and live cells.
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DOI：
10.1126/science.aaq1392
发表时间：
2018-04-20
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Liu TL;Upadhyayula S;Milkie DE;Singh V;Wang K;Swinburne IA;Mosaliganti KR;Collins ZM;Hiscock TW;Shea J;Kohrman AQ;Medwig TN;Dambournet D;Forster R;Cunniff B;Ruan Y;Yashiro H;Scholpp S;Meyerowitz EM;Hockemeyer D;Drubin DG;Martin BL;Matus DQ;Koyama M;Megason SG;Kirchhausen T;Betzig E
通讯作者：
Betzig E
影响因子：
48
作者：
Tomer, Raju;Khairy, Khaled;Keller, Philipp J.
通讯作者：
Keller, Philipp J.
影响因子：
3.8
作者：
Chang, Bo-Jui;Dean, Kevin M.;Fiolka, Reto
通讯作者：
Fiolka, Reto
影响因子：
7.7
作者：
Boothe, Tobias;Hilbert, Lennart;Rink, Jochen C.
通讯作者：
Rink, Jochen C.
影响因子：
48
作者：
Grimm JB;Muthusamy AK;Liang Y;Brown TA;Lemon WC;Patel R;Lu R;Macklin JJ;Keller PJ;Ji N;Lavis LD
通讯作者：
Lavis LD