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Frontiers of Optoelectronics

Front. Optoelectron.    2015, Vol. 8 Issue (2) : 177-182     DOI: 10.1007/s12200-015-0503-1
Fluorescence microendoscopy imaging based on GRIN lenses with one- and two-photon excitation modes
Wei YAN1,2,Xiao PENG1,Danying LIN1,Qi WANG1,Jian GAO1,Teng LUO1,Jie ZHOU1,Tong YE2,*(),Junle QU1,*(),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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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 Authors: 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.
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Danying LIN
Jian GAO
Teng LUO
Tong YE
Junle QU
Hanben NIU
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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