Self‐referenced axial chromatic dispersion measurement in multiphoton microscopy through 2‐color third‐harmonic generation imaging. Issue 9 (8th June 2018)

• Record Type: Journal Article
• Authors: Du, Yu; Zhuang, Ziwei; He, Jiexing; Liu, Hongji; Qiu, Ping; Wang, Ke
• Is Part Of: Journal of biophotonics. Volume 11:Issue 9(2018)
• Page Start: n/a
• Page End: n/a
• Publication Date: 2018-06-08
• Subjects: 2‐color -- axial chromatic dispersion -- multiphoton microscopy; Photonics -- Periodicals; Optical materials -- Periodicals; Optics -- Periodicals; Medical instruments and apparatus -- Periodicals; 621.3605
• DOI: 10.1002/jbio.201800071
• Languages: English
• ISSNs: 1864-063X

Abstract

Abstract : With tunable excitation light, multiphoton microscopy is widely used for imaging biological structures at subcellular resolution. Axial chromatic dispersion, present in virtually every transmissive optical system including the multiphoton microscope, leads to focal (and the resultant image) plane separation. Here, we experimentally demonstrate a technique to measure the axial chromatic dispersion in a multiphoton microscope, using simultaneous 2‐color third‐harmonic generation imaging excited by a 2‐color soliton source with tunable wavelength separation. Our technique is self‐referenced, eliminating potential measurement error when 1‐color tunable excitation light is used which necessitates reciprocating motion of the mechanical translation stage. Using this technique, we demonstrate measured axial chromatic dispersion with 2 different objective lenses in a multiphoton microscope. Further measurement in a biological sample also indicates that this axial chromatic dispersion, in combination with 2‐color imaging, may open up opportunity for simultaneous imaging of 2 different axial planes. Abstract : This study demonstrates self‐referenced axial chromatic dispersion measurement in multiphoton microscopy through simultaneous 2‐color THG imaging. A 2‐color soliton source was built as the excitation source, with each soliton wavelength continuously tunable. Using this technique, axial chromatic dispersions of different objective lenses are shown. Furthermore, 2‐color THG imaging in a biological sample using the dispersive multiphoton microscope demonstrates its potential applicability to visualizing two different axial layers simultaneously.