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Frequency domain optical resolution photoacoustic and fluorescence microscopy using a modulated laser diode

Langer, Gregor ; Langer, Andreas LU orcid ; Buchegger, Bianca ; Jacak, Jaroslaw ; Klar, Thomas A. and Berer, Thomas (2017) Photons Plus Ultrasound: Imaging and Sensing 2017 In Progress in Biomedical Optics and Imaging - Proceedings of SPIE 10064.
Abstract

In this paper a multimodal optical-resolution photoacoustic and fluorescence microscope in frequency domain is presented. Photoacoustic waves and modulated fluorescence are generated in chromophores by using a modulated diode laser. The photoacoustic waves, recorded with a hydrophone, and the fluorescence signals, acquired with an avalanche photodiode, are simultaneously measured using a lock-in technique. Two possibilities to optimize the signal-to-noise ratio are discussed. The first method is based on the optimization of the excitation waveform and it is argued why square-wave excitation is best. The second way to enhance the SNR is to optimize the modulation frequency. For modulation periods that are much shorter than the relaxation... (More)

In this paper a multimodal optical-resolution photoacoustic and fluorescence microscope in frequency domain is presented. Photoacoustic waves and modulated fluorescence are generated in chromophores by using a modulated diode laser. The photoacoustic waves, recorded with a hydrophone, and the fluorescence signals, acquired with an avalanche photodiode, are simultaneously measured using a lock-in technique. Two possibilities to optimize the signal-to-noise ratio are discussed. The first method is based on the optimization of the excitation waveform and it is argued why square-wave excitation is best. The second way to enhance the SNR is to optimize the modulation frequency. For modulation periods that are much shorter than the relaxation times of the excited chromophores, the photoacoustic signal scales linearly with the modulation frequency. We come to the conclusion that frequency-domain photoacoustic microscopy performed with modulation frequencies in the range of 100 MHz can compete with time-domain photoacoustic microscopy regarding the signal-to-noise ratio. The theoretical predictions are confirmed by experimental results. Additionally, images of stained and unstained biological samples are presented in order to demonstrate the capabilities of the multimodal imaging system.

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author
; ; ; ; and
publishing date
type
Chapter in Book/Report/Conference proceeding
publication status
published
subject
keywords
Diode laser, Fluorescence microscopy, Frequency domain, Lock-in technique, Multimodal imaging, Optical resolution photoacoustic microscopy, Photoacoustic microscopy, Square wave excitation
host publication
Photons Plus Ultrasound : Imaging and Sensing 2017 - Imaging and Sensing 2017
series title
Progress in Biomedical Optics and Imaging - Proceedings of SPIE
editor
Oraevsky, Alexander A. and Wang, Lihong V.
volume
10064
article number
1006426
publisher
SPIE
conference name
Photons Plus Ultrasound: Imaging and Sensing 2017
conference location
San Francisco, United States
conference dates
2017-01-29 - 2017-02-01
external identifiers
  • scopus:85018926035
ISSN
1605-7422
ISBN
9781510605695
DOI
10.1117/12.2250861
language
English
LU publication?
no
additional info
Publisher Copyright: © 2017 SPIE. Copyright: Copyright 2018 Elsevier B.V., All rights reserved.
id
62c3d921-f8f9-4a8a-94e8-ce2833d2db9e
date added to LUP
2021-03-15 22:33:43
date last changed
2024-07-13 11:09:06
@inproceedings{62c3d921-f8f9-4a8a-94e8-ce2833d2db9e,
  abstract     = {{<p>In this paper a multimodal optical-resolution photoacoustic and fluorescence microscope in frequency domain is presented. Photoacoustic waves and modulated fluorescence are generated in chromophores by using a modulated diode laser. The photoacoustic waves, recorded with a hydrophone, and the fluorescence signals, acquired with an avalanche photodiode, are simultaneously measured using a lock-in technique. Two possibilities to optimize the signal-to-noise ratio are discussed. The first method is based on the optimization of the excitation waveform and it is argued why square-wave excitation is best. The second way to enhance the SNR is to optimize the modulation frequency. For modulation periods that are much shorter than the relaxation times of the excited chromophores, the photoacoustic signal scales linearly with the modulation frequency. We come to the conclusion that frequency-domain photoacoustic microscopy performed with modulation frequencies in the range of 100 MHz can compete with time-domain photoacoustic microscopy regarding the signal-to-noise ratio. The theoretical predictions are confirmed by experimental results. Additionally, images of stained and unstained biological samples are presented in order to demonstrate the capabilities of the multimodal imaging system.</p>}},
  author       = {{Langer, Gregor and Langer, Andreas and Buchegger, Bianca and Jacak, Jaroslaw and Klar, Thomas A. and Berer, Thomas}},
  booktitle    = {{Photons Plus Ultrasound : Imaging and Sensing 2017}},
  editor       = {{Oraevsky, Alexander A. and Wang, Lihong V.}},
  isbn         = {{9781510605695}},
  issn         = {{1605-7422}},
  keywords     = {{Diode laser; Fluorescence microscopy; Frequency domain; Lock-in technique; Multimodal imaging; Optical resolution photoacoustic microscopy; Photoacoustic microscopy; Square wave excitation}},
  language     = {{eng}},
  publisher    = {{SPIE}},
  series       = {{Progress in Biomedical Optics and Imaging - Proceedings of SPIE}},
  title        = {{Frequency domain optical resolution photoacoustic and fluorescence microscopy using a modulated laser diode}},
  url          = {{http://dx.doi.org/10.1117/12.2250861}},
  doi          = {{10.1117/12.2250861}},
  volume       = {{10064}},
  year         = {{2017}},
}