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Multimodal spectral focusing CARS and SFG microscopy with a tailored coherent continuum from a microstructured fiber

Multimodal spectral focusing CARS and SFG microscopy with a tailored coherent continuum from a microstructured fiber
Multimodal spectral focusing CARS and SFG microscopy with a tailored coherent continuum from a microstructured fiber
We report a technologically novel microscopy system for bioimaging based on 100 fs Titanium:Sapphire (Ti:Sa) laser coherent continuum from a tailored, 9 cm long All Normal Dispersion (ANDi) fiber, enabling concurrent image contrast with a) Spectral Focusing Coherent Anti-Stokes Raman Scattering (SF-CARS) (spanning 900-3200 cm-1) and b) sum-frequency-generation (SFG). Both modalities were efficiently excited with power levels at the microscope focus compatible with biological samples. Moreover, using the continuum, images were recorded in the back-scattering (epi-detection) geometry, without the necessity for an expensive, computer-controlled, Spatial Light Modulator (SLM), clearly demonstrating the strong signal levels achieved. Image contrast from the multiple modalities provided greater chemical and structural insights than imaging with any single technique in isolation. Numerical simulations supported these developments in regard to both the optimum fiber length for SC generation and the achievement of high spectral resolution in SF-CARS via careful group-delay-dispersion matching across the pump and Stokes pulses using just an inexpensive sequence of short glass blocks inserted into the Stokes beam. We show bio-images of mouse tissue recorded concurrently via label/stain-free contrast from multiple modalities: CARS, Two Photon auto-Fluorescence (TPaF) and Second Harmonic/Sum Frequency Generation (SHG/SFG). Overall our approach delivers optimum performance in back-scattered (epi-) detection configuration, suited for thick samples, at reduced complexity and cost. The addition of this simple fiber add-on to lasers already widely used for TPF microscopy can thus extend the capabilities of a significant number of existing microscopy laboratories.
0946-2171
Herdzik, Krzysztof, Pawel
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Bourdakos, Konstantinos
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Johnson, Peter
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Lister, Adam, Paul
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Pitera, Aleksandra, Paulina
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Guo, Chun-Yu
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Horak, Peter
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Richardson, David
ebfe1ff9-d0c2-4e52-b7ae-c1b13bccdef3
Price, Jonathan
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Mahajan, Sumeet
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Herdzik, Krzysztof, Pawel
c94de69f-61c5-4ba0-bed5-dd13c5d9ec0b
Bourdakos, Konstantinos
83f6fc3a-db12-476b-9a78-4aad8756f82f
Johnson, Peter
862b092d-ac8b-4c9b-804f-10d42d8b0d21
Lister, Adam, Paul
a79ccf69-a17b-4d0a-9fe2-28287e3eddf3
Pitera, Aleksandra, Paulina
6cf28d67-483b-494b-8323-c01a64c24f71
Guo, Chun-Yu
9d003bc2-a710-402d-b59c-3c8cdf126b5f
Horak, Peter
520489b5-ccc7-4d29-bb30-c1e36436ea03
Richardson, David
ebfe1ff9-d0c2-4e52-b7ae-c1b13bccdef3
Price, Jonathan
fddcce17-291b-4d01-bd38-8fb0453abdc8
Mahajan, Sumeet
b131f40a-479e-4432-b662-19d60d4069e9

Herdzik, Krzysztof, Pawel, Bourdakos, Konstantinos, Johnson, Peter, Lister, Adam, Paul, Pitera, Aleksandra, Paulina, Guo, Chun-Yu, Horak, Peter, Richardson, David, Price, Jonathan and Mahajan, Sumeet (2020) Multimodal spectral focusing CARS and SFG microscopy with a tailored coherent continuum from a microstructured fiber. Applied Physics B, 126, [84]. (doi:10.1007/s00340-020-7406-6).

Record type: Article

Abstract

We report a technologically novel microscopy system for bioimaging based on 100 fs Titanium:Sapphire (Ti:Sa) laser coherent continuum from a tailored, 9 cm long All Normal Dispersion (ANDi) fiber, enabling concurrent image contrast with a) Spectral Focusing Coherent Anti-Stokes Raman Scattering (SF-CARS) (spanning 900-3200 cm-1) and b) sum-frequency-generation (SFG). Both modalities were efficiently excited with power levels at the microscope focus compatible with biological samples. Moreover, using the continuum, images were recorded in the back-scattering (epi-detection) geometry, without the necessity for an expensive, computer-controlled, Spatial Light Modulator (SLM), clearly demonstrating the strong signal levels achieved. Image contrast from the multiple modalities provided greater chemical and structural insights than imaging with any single technique in isolation. Numerical simulations supported these developments in regard to both the optimum fiber length for SC generation and the achievement of high spectral resolution in SF-CARS via careful group-delay-dispersion matching across the pump and Stokes pulses using just an inexpensive sequence of short glass blocks inserted into the Stokes beam. We show bio-images of mouse tissue recorded concurrently via label/stain-free contrast from multiple modalities: CARS, Two Photon auto-Fluorescence (TPaF) and Second Harmonic/Sum Frequency Generation (SHG/SFG). Overall our approach delivers optimum performance in back-scattered (epi-) detection configuration, suited for thick samples, at reduced complexity and cost. The addition of this simple fiber add-on to lasers already widely used for TPF microscopy can thus extend the capabilities of a significant number of existing microscopy laboratories.

Text
20190805_SF-CARS ANDi fibre - revision vfinal - Accepted Manuscript
Restricted to Repository staff only until 21 February 2021.
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Accepted/In Press date: 22 February 2020
e-pub ahead of print date: 17 April 2020

Identifiers

Local EPrints ID: 438391
URI: http://eprints.soton.ac.uk/id/eprint/438391
ISSN: 0946-2171
PURE UUID: 3c6a240c-1f9f-4984-9720-d15d8f033f97
ORCID for Peter Horak: ORCID iD orcid.org/0000-0002-8710-8764
ORCID for David Richardson: ORCID iD orcid.org/0000-0002-7751-1058
ORCID for Jonathan Price: ORCID iD orcid.org/0000-0003-0256-9172
ORCID for Sumeet Mahajan: ORCID iD orcid.org/0000-0001-8923-6666

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Date deposited: 09 Mar 2020 17:30
Last modified: 19 May 2020 00:36

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