Full Text:   <1755>

Summary:  <882>

CLC number: O439

On-line Access: 2021-10-08

Received: 2020-08-12

Revision Accepted: 2020-11-06

Crosschecked: 2020-12-24

Cited: 0

Clicked: 2866

Citations:  Bibtex RefMan EndNote GB/T7714

 ORCID:

Ruheng Shi

https://orcid.org/0000-0001-7993-708X

Lingjie Kong

https://orcid.org/0000-0002-8250-7547

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Frontiers of Information Technology & Electronic Engineering  2021 Vol.22 No.10 P.1289-1298

http://doi.org/10.1631/FITEE.2000410


Enhanced collection of scattered photons in nonlinear fluorescence microscopy by extended epi-detection with a silicon photomultiplier array


Author(s):  Ruheng Shi, Cheng Jin, Chi Liu, Lingjie Kong

Affiliation(s):  State Key Laboratory of Precision Measurement Technology and Instruments, Department of Precision Instrument, Tsinghua University, Beijing 100084, China; more

Corresponding email(s):   konglj@tsinghua.edu.cn

Key Words:  Extended epi-detection, Enhanced collection, Nonlinear fluorescence microscopy, Silicon photomultiplier array


Ruheng Shi, Cheng Jin, Chi Liu, Lingjie Kong. Enhanced collection of scattered photons in nonlinear fluorescence microscopy by extended epi-detection with a silicon photomultiplier array[J]. Frontiers of Information Technology & Electronic Engineering, 2021, 22(10): 1289-1298.

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publisher="Zhejiang University Press & Springer",
doi="10.1631/FITEE.2000410"
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Abstract: 
To maximize signal collection in nonlinear optical microscopy, non-descanned epi-detection is generally adopted for in vivo imaging. However, because of severe scattering in biological samples, most of the emitted fluorescence photons go beyond the collection angles of objectives and thus cannot be detected. Here, we propose an extended detection scheme to enhance the collection of scattered photons in nonlinear fluorescence microscopy using a silicon photomultiplier array ahead of the front apertures of objectives. We perform numerical simulations to demonstrate the enhanced fluorescence collection via extended epi-detection in the multi-photon fluorescence imaging of human skin and mouse brain through craniotomy windows and intact skulls. For example, with red fluorescence emission at a depth of 600 μm in human skin, the increased collection can be as much as about 150% with a 10×, 0.6-NA objective. We show that extended epi-detection is a generally applicable, feasible technique for use in nonlinear fluorescence microscopy to enhance signal detection.

采用硅光电倍增管阵列扩展探测实现非线性荧光显微技术中散射光子探测信号的增强

施汝恒1,靳程1,刘驰1,孔令杰1,2
1清华大学精密仪器系精密测试技术与仪器国家重点实验室,中国北京市,100084
2清华大学IDG/麦戈文脑科学研究院,中国北京市,100084
摘要:为提高非线性荧光显微技术在活体成像中的信号探测能力,人们常采用基于非退扫描的背向探测策略。然而,由于生物样本中存在严重散射,物镜前孔径处大部分荧光光子辐射角度超出物镜收集角范围,因而无法被探测。本文提出一种将硅光电倍增管阵列放置在物镜前孔径处以增强非线性显微镜对散射光子探测能力的扩展探测方案。通过数值仿真说明了扩展探测在对人体皮肤和小鼠大脑(透过颅窗及完整颅骨)进行多光子荧光成像的信号增强情况。例如,在人类皮肤60 µm成像深度处使用红色染料标记情况下,使用10×,0.6NA物镜进行成像,扩展探测方案引入的信号增强可达约150%。本文论证了扩展探测是一种灵活、广泛适用于非线性荧光显微镜增强探测信号的技术。

关键词:扩展背向探测;增强探测;非线性荧光显微镜;硅光电倍增管阵列

Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article

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