Super Resolution Measurement of Collagen Fibers in Biological Samples: Validation of a Commercial Solution for Multiphoton Microscopy

Overview

Journal PLoS One

Specialties General Medicine
Science

Date 2020 Feb 15

PMID 32059025

Citations 9

Authors

Aaron M Barlow

Leila B Mostaco-Guidolin

Emmanuel T Osei

Steven Booth

Tillie-Louise Hackett

Affiliations

Soon will be listed here.

Abstract

Multiphoton microscopy is a powerful, non-invasive technique to image biological specimens. One current limitation of multiphoton microscopy is resolution as many of the biological molecules and structures investigated by research groups are similar in size or smaller than the diffraction limit. To date, the combination of multiphoton and super-resolution imaging has proved technically challenging for biology focused laboratories to implement. Here we validate that the commercial super-resolution Airyscan detector from ZEISS, which is based on image scanning microscopy, can be integrated under warranty with a pulsed multi-photon laser to enable multiphoton microscopy with super-resolution. We demonstrate its biological application in two different imaging modalities, second harmonic generation (SHG) and two-photon excited fluorescence (TPEF), to measure the fibre thicknesses of collagen and elastin molecules surpassing the diffraction limit by a factor of 1.7±0.3x and 1.4±0.3x respectively, in human heart and lung tissues, and 3-dimensional in vitro models. We show that enhanced resolution and signal-to-noise of SHG using the Airyscan compared to traditional GaAs detectors allows for automated and precise measurement of collagen fibres using texture analysis in biological tissues.

Citing Articles

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