Development and characterization of a two-photon fluorescence microscope for collagen tissue based on a nanosecond laser

Tobias Baselt; Robert Scheller; Burkhard Fleck; Peter Hartmann

doi:10.1117/12.2673661

11 August 2023 Development and characterization of a two-photon fluorescence microscope for collagen tissue based on a nanosecond laser

Tobias Baselt, Robert Scheller, Burkhard Fleck, Peter Hartmann

Author Affiliations +

Proceedings Volume 12622, Optical Methods for Inspection, Characterization, and Imaging of Biomaterials VI; 126220H (2023) https://doi.org/10.1117/12.2673661
Event: SPIE Optical Metrology, 2023, Munich, Germany

Abstract

The structural and microscopic examination of cartilage and tendon tissue is of central interest for the characterization of torn ligament healing or pathological tissue changes. Conventional microscopy or optical coherence tomography methods can only partially resolve tissue types and their structural orientation. Two or multiphoton microscopy and second harmonic generation provide information about the material composition or alignment of nanoscaled structural proteins. In most systems, femtosecond lasers are used, which leads to high system costs and requires special system components. The aim of this work is to investigate a narrowband nanosecond laser with a pulse energy several orders of magnitude higher, reducing the required number of laser pulses to be averaged per pixel. Second harmonic generation and two photon excited fluorescence can be used to non-invasively examine deeper tissue structures. High lateral resolution was achieved by scanning the sample. Simultaneous real-time visualization of collagenous and cellular structures was attained. Defined aligned collagenous fibers of a sheep tendon were investigated. The anisotropy of the collagenous structures could be demonstrated. It was possible to realize a two-dimensional imaging method with a maximum point density of 5080 PPI and a numerical aperture of 0.15. The method allows the simultaneous separate observation of collagen fibers through the use of second harmonic generation and cellular tissue using a narrow-band nanosecond laser for two photon excited fluorescence.

Conference Presentation

Citation Download Citation

Tobias Baselt, Robert Scheller, Burkhard Fleck, and Peter Hartmann "Development and characterization of a two-photon fluorescence microscope for collagen tissue based on a nanosecond laser", Proc. SPIE 12622, Optical Methods for Inspection, Characterization, and Imaging of Biomaterials VI, 126220H (11 August 2023); https://doi.org/10.1117/12.2673661

ACCESS THE FULL ARTICLE

INSTITUTIONAL
Select your institution to access the SPIE Digital Library.

SELECT YOUR INSTITUTION

PERSONAL
Sign in with your SPIE account to access your personal subscriptions or to use specific features such as save to my library, sign up for alerts, save searches, etc.

PERSONAL SIGN IN

No SPIE Account? Create one

PURCHASE THIS CONTENT

SUBSCRIBE TO DIGITAL LIBRARY

50 downloads per 1-year subscription

Members: $195

Non-members: $335 ADD TO CART

25 downloads per 1 - year subscription

Members: $145

Non-members: $250 ADD TO CART

PURCHASE SINGLE ARTICLE

Includes PDF, HTML & Video, when available