Reflective ultrathin light-sheet microscopy with isotropic 3D resolutions

Yue Wang; Dashan Dong; Wenkai Yang; Renxi He; Ming Lei; Kebin Shi

doi:10.1364/PRJ.500618

Journals >Photonics Research >Volume 12 >Issue 2 >Page 271 > Article

Photonics Research
Vol. 12, Issue 2, 271 (2024)

Reflective ultrathin light-sheet microscopy with isotropic 3D resolutions | Spotlight on Optics

Yue Wang¹, Dashan Dong¹, Wenkai Yang¹, Renxi He², Ming Lei³, and Kebin Shi^{1、4、5、*}

Author Affiliations

¹State Key Laboratory for Mesoscopic Physics, Department of Physics, Peking University, Beijing 100871, China

²State Key Laboratory of Membrane Biology, School of Life Sciences, and Biomedical Pioneering Innovation Center (BIOPIC), Peking University, Beijing 100871, China

³School of Physics, Xi’an Jiaotong University, Xi’an 710049, China

⁴Collaborative Innovation Center of Extreme Optics, Shanxi University, Taiyuan 030006, China

⁵Peking University Yangtze Delta Institute of Optoelectronics, Nantong 226010, China

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DOI: 10.1364/PRJ.500618 Cite this Article Set citation alerts

Yue Wang, Dashan Dong, Wenkai Yang, Renxi He, Ming Lei, Kebin Shi. Reflective ultrathin light-sheet microscopy with isotropic 3D resolutions[J]. Photonics Research, 2024, 12(2): 271 Copy Citation Text

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Abstract

Light-sheet fluorescence microscopy (LSFM) has played an important role in bio-imaging due to its advantages of high photon efficiency, fast speed, and long-term imaging capabilities. The perpendicular layout between LSFM excitation and detection often limits the 3D resolutions as well as their isotropy. Here, we report on a reflective type light-sheet microscope with a mini-prism used as an optical path reflector. The conventional high NA objectives can be used both in excitation and detection with this design. Isotropic resolutions in 3D down to 300 nm could be achieved without deconvolution. The proposed method also enables easy transform of a conventional fluorescence microscope to high performance light-sheet microscopy.

\nabla^{2} E (\vec{r}) + \frac{4 π^{2}}{λ_{0}^{2}} n^{2} (\vec{r}) E (\vec{r}) = 0 .

(1)

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E (\vec{r}) = U (\vec{r}) \exp (j \frac{2 π}{λ_{0}} n_{0} z),

(2)

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U_{k + 1} = \exp [\frac{j λ_{0} d z}{4 π n_{0}} \nabla_{⊥} + \frac{j 2 π}{λ_{0}} δ n (\vec{r}) d z] U_{k} .

(3)

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U_{k + 1} = [\frac{j 2 π}{λ_{0}} δ n (\vec{r}) d z] \times IFT {FT {U_{k}} \exp [- \frac{j π λ_{0} d z}{n_{0}} (f_{x}^{2} + f_{y}^{2})]} .

(4)

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U_{k + 1} = [\frac{j 2 π}{λ_{0}} δ n (\vec{r}) d z] \times IFT {FT {U_{k}} \exp [- \frac{j λ_{0} d z}{2 π n_{0} + \sqrt{4 π^{2} n_{0}^{2} - λ_{0}^{2} (f_{x}^{2} + f_{y}^{2})}} (f_{x}^{2} + f_{y}^{2})]} .

(5)

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{PSF}_{o} (x, y, z) = {PSF}_{e} (x, y, z) \times {PSF}_{d} (x, y, z) .

(6)

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{PSF}_{o} (x, y, z) = I_{e} (x, y, z) \times {PSF}_{d} (x, y, z) .

(7)

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2 z_{0} = \frac{2 π n ω_{0}^{2}}{λ} .

(C1)

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Yue Wang, Dashan Dong, Wenkai Yang, Renxi He, Ming Lei, Kebin Shi. Reflective ultrathin light-sheet microscopy with isotropic 3D resolutions[J]. Photonics Research, 2024, 12(2): 271

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