Fluorescence microendoscopy imaging based on GRIN lenses with one- and two-photon excitation modes

doi:10.1007/s12200-015-0503-1

Front. Optoelectron.

2015, Vol. 8

Issue (2) : 177-182 https://doi.org/10.1007/s12200-015-0503-1

RESEARCH ARTICLE

Fluorescence microendoscopy imaging based on GRIN lenses with one- and two-photon excitation modes

Wei YAN^1,²,Xiao PENG¹,Danying LIN¹,Qi WANG¹,Jian GAO¹,Teng LUO¹,Jie ZHOU¹,Tong YE^2,^*(

),Junle QU^1,^*(

),Hanben NIU¹

1. Key Laboratory of Optoelectronic Devices and Systems of Guangdong Province, College of Optoelectronic Engineering, Shenzhen University, Shenzhen 518060, China
2. Department of Bioengineering, Clemson University, CU-MUSC Bioengineering Program, Charleston 29425, USA

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Abstract

With the rapid development of life sciences, there is an increasing demand for intravital fluorescence imaging of small animals. However, large dimensions and limited working distances of objective lenses in traditional fluorescence microscopes have limited their imaging applications mostly to superficial tissues. To overcome these disadvantages, researchers have developed the graded-index (GRIN) probes with small diameters for imaging internal organs of small animals in a minimally invasive fashion. However, dynamic imaging based on GRIN lens has not been studied extensively. Here, this paper presented a fluorescence endoscopic imaging system based on GRIN lenses using one-photon and two-photon excitation. GRIN lenses with 1.15 mm diameter and 7.65 mm length were used in the system. The images were acquired by a compact laser scanning imaging system with a resonant galvo-mirror system to scan the laser beam and a photomultiplier tube (PMT) to detect fluorescence signals. Experimental results showed that this system using two-photon excitation could implement dynamic fluorescence microendoscopic imaging and monitor the movement of blood flow beneath the skin in anesthetized mice while producing images with higher contrast and signal to noise ratio (SNR) than those using one photon excitation. It would be a useful tool for studying biological processes of small animals or plants in vivo.

Keywords one-photon excitation two-photon excitation graded-index (GRIN) lens fluorescence micorendoscopy dynamic imaging

Corresponding Author(s): Tong YE,Junle QU

Just Accepted Date: 15 May 2015 Online First Date: 01 June 2015 Issue Date: 24 June 2015

Cite this article:

Wei YAN,Xiao PENG,Danying LIN, et al. Fluorescence microendoscopy imaging based on GRIN lenses with one- and two-photon excitation modes[J]. Front. Optoelectron., 2015, 8(2): 177-182.

URL:

https://academic.hep.com.cn/foe/EN/10.1007/s12200-015-0503-1
https://academic.hep.com.cn/foe/EN/Y2015/V8/I2/177

Fig.1 Experimental setup of the fluorescence microendoscopy imaging system using GRIN lenses. The right panel is the GRIN lens and its parameters. Laser: Spectra-Physics, Mai Tai DeepSee, or a semiconductor laser; DM: dichroic mirror; F: blocking filter; PMT: photomultiplier tube, Hamamatsu H7422-40; Objective: Olympus 10 × /NA 0.3 dry objective

Fig.2 One-photon excited fluorescence microendoscopic images of cells in (a) a Convallaria rhizome slide; (b) inside of an Aloe stem; (c) normal breast cells and (d) breast cancer cells in slides

Fig.3 Two-photon excited fluorescence images of cells in (a) a Convallaria rhizome slide; (b) inside of an Aloe stem

Fig.4 Dynamic fluorescence images (imaging speed 20 frames/s, 512 × 512 pixels per frame) of blood flow beneath the skin of an anesthetized mouse. The blood flow was enabled to be visible by staining with fluorescein isothiocyanate-dextran (average mol wt 2000000). The images were captured at t = 0 s (a), 0.05 s (b), 0.10 s (c) and 0.15 s (d), respectively. The marked red spot is an aggregate of the dye whose movement was clearly observed during the dynamic imaging process

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