Design of a fiber-optic multiphoton microscopy handheld probe.

Yuan Zhao,Lin Huang,Mingyu Sheng,Shuo Tang

doi:10.1364/boe.7.003425

Abstract

We have developed a fiber-optic multiphoton microscopy (MPM) system with handheld probe using femtosecond fiber laser. Here we present the detailed optical design and analysis of the handheld probe. The optical systems using Lightpath 352140 and 352150 as objective lens were analyzed. A custom objective module that includes Lightpath 355392 and two customized corrective lenses was designed. Their performances were compared by wavefront error, field curvature, astigmatism, F-θ error, and tolerance in Zemax simulation. Tolerance analysis predicted the focal spot size to be 1.13, 1.19 and 0.83 µm, respectively. Lightpath 352140 and 352150 were implemented in experiment and the measured lateral resolution was 1.22 and 1.3 µm, respectively, which matched with the prediction. MPM imaging by the handheld probe were conducted on leaf, fish scale and rat tail tendon. The MPM resolution can potentially be improved by the custom objective module.

Highlights

Multiphoton microscopy (MPM) is an important biological imaging modality
Requirement to expand the applications of MPM by handheld probe or endoscope is urgent, where light can be delivered in flexible fiber and images can be captured using a miniaturized probe
Two-photon excited fluorescence (TPEF) images of 0.1-μm diameter fluorescent beads (Fluoresbrite Calibration Grade Size Range Kit, Polysciences, Inc.) are acquired. 2D image is acquired by scanning the MEMS mirror in X and Y directions

Summary

Introduction

Multiphoton microscopy (MPM) is an important biological imaging modality. It can be applied to non-invasive study of biological samples to obtain sub-micron resolution imaging in three dimensions. MPM utilizes multiple incident photons to excite one emission photon. This nonlinear imaging modality generates sharp contrast and high resolution images [1,2,3,4,5]. Requirement to expand the applications of MPM by handheld probe or endoscope is urgent, where light can be delivered in flexible fiber and images can be captured using a miniaturized probe. To meet the above requirement, many research groups focus on endoscope design and establishment [6,7,8,9]

Methods

Results

Conclusion