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Speckle Correlation and Stimulated Raman Scattering: towards Species-Specific 3D Imaging
Luleå University of Technology, Department of Engineering Sciences and Mathematics, Fluid and Experimental Mechanics.ORCID iD: 0000-0002-0306-3010
2025 (English)Doctoral thesis, comprehensive summary (Other academic)Alternative title
Specklekorrelation och stimulerad Raman spridning: mot ämnesspecifik 3D avbildning (Swedish)
Abstract [en]

Light can be used in a wide range of measurement methods, from probing the structure of a molecule to quality control of industrial-produced products. This thesis is the result of the initial work connected to a research project regarding the development of interferometric imaging of stimulated Raman scattering (InFeRa). The combination of two different imaging methods, interferometry and stimulated Raman scattering, into one measurement system would provide images containing information about a sample's three-dimensional structure together with chemical specificity. This would allow for studies of a sample's chemical and structural changes due to environmental stimuli to be investigated simultaneously. 

Raman spectroscopy is a label-free, non-contact, and non-destructive optical measurement method based on inelastic scattering of light from a material illuminated by a laser. The Raman scattered light is shown in a Raman spectrum that provides information about the material's molecular structure. This allows for precise identification of samples and investigation of changes in the molecular structure and bond strength. In \textbf{Paper Paper A, Raman spectroscopy was used to investigate bacterial nano cellulose (BNC) produced by bacteria feed with three different carbon sources: glucose, terephthalic acid (TPA), and ethylene glycol (EG). The results were compared to measurements of commercially produced cellulose from plant matter. The investigation showed changes to the glycidic bond responsible for connecting glucose rings with cellulose chains. 

Stimulated Raman scattering (SRS) is a powerful imaging technique that has become popular during the last decades for its ability to image specific species in a sample with high accuracy. When a sample's Raman spectrum is known, SRS can be used to image targeted molecular vibrations in a sample to distinguish between different compounds or to track molecular changes due to environmental stimuli. In SRS, two laser beams, called the Stokes and the pump beams, respectively, are used. SRS can be achieved if the wavelength difference between the two laser beams corresponds to a Raman molecular vibration mode. SRS images are today commonly produced using scanning microscopes, limiting the type of samples that can be imaged.  Implementing an SRS imaging system without relying on high-numerical-aperture microscope objectives and scanning stages would enable the investigation of samples that do not fit under a microscope. If SRS imaging is to be realized in 3D using cameras, understanding the 3D spatial generation of the signal is important. 

It is also of interest to be able to control where the signal is generated in the sample. The working principle of the experiments was to illuminate a cylindrical volume of the sample with the Stokes beam and to use a lens to focus the pump beam into this volume. In Paper B the spatial generation in ethanol was investigated by experiments and simulations. A dichroic mirror was inserted into the ethanol sample to separate the pump, and Stokes beam at different positions, thus stopping the generation of SRS and allowing the resulting signal to be studied. Further, the pump intensity was varied to study the effect on the spatial width of the SRS signal. The experimental results were further compared to computer simulations. The simulations were based on diffraction theory for the beam propagation, and the interaction between the light beams and the material was modeled with a phase modulation due to the induced Kerr effect caused by the high pump intensity. The results show that most of the SRS generation takes place close to the focus of the pump beam. In Conference contribution A, a phase spatial light modulator (SLM) was integrated into the SRS experimental setup to allow for spatial control of the generated SRS signal. It was shown that the shape and position of the pump beam focus and, thereby, the shape of the generated SRS signal could be controlled.

Interferometric imaging can provide information on a sample's 3D structure in various applications. However, an alternative approach was explored due to certain limitations and requirements of SRS imaging.SRS imaging with cameras requires the capturing of two images: one containing the SRS signal and a reference image without the signal. Ideally, these images should be recorded simultaneously, which is not possible with the system used in Paper B and Conference contribution A. Another limitation is the sensitivity of an interferometric imaging system to disturbances. To address these limitations, a robust dual-camera imaging system that provides phase contrast imaging based on digital image correlation of speckles was demonstrated in Paper C. The system utilizes the light deflection in an image's Fourier plane, which has a linear relation to the refraction strength of a phase object and can be calculated from the speckle correlation between the two cameras. The phase object was reconstructed using digital image correlation for a large number of independent speckle patterns. The system was evaluated with experiments and simulations. The system was shown to produce a quantitative measurement of the phase objectives' refraction strength with sensitivity on par with digital holographic interference measurements. However, some errors in the reconstruction were identified to be caused by an out-of-plane movement of the speckle field. 

The image quality of the reconstructed image is internally linked to the standard deviation in the correlation value, and the number of images used in the image reconstruction, which is investigated in Papper D. The standard deviation in the correlation values for different numbers of images used in the reconstruction was investigated for different f-numbers with experiments and simulations. The results show that the f-number of the camera objective controls the imaging system sensitivity but does not directly impact the precision of the measurements. The standard deviation drops off as 1/N1/2 where is the number of images used in the reconstruction.

Place, publisher, year, edition, pages
Luleå: Luleå University of Technology, 2025.
Series
Doctoral thesis / Luleå University of Technology, ISSN 1402-1544
Keywords [en]
Stimulated Raman scattering, phase contrast imaging, spatial light modulator, speckle correlation imaging, bacterial nanocellulosa, imaging
National Category
Atom and Molecular Physics and Optics
Research subject
Experimental Mechanics
Identifiers
URN: urn:nbn:se:ltu:diva-112186ISBN: 978-91-8048-805-1 (print)ISBN: 978-91-8048-806-8 (electronic)OAI: oai:DiVA.org:ltu-112186DiVA, id: diva2:1948778
Public defence
2025-06-04, E632, Luleå University of Technology, Luleå, 09:00 (English)
Opponent
Supervisors
Available from: 2025-04-01 Created: 2025-03-31 Last updated: 2025-10-21Bibliographically approved
List of papers
1. Characterization of bacterial nanocellulose cultivated on polyethylene terephthalate (PET) monomers via raman and fourier transform infrared spectroscopy
Open this publication in new window or tab >>Characterization of bacterial nanocellulose cultivated on polyethylene terephthalate (PET) monomers via raman and fourier transform infrared spectroscopy
2026 (English)In: Scientific Reports, E-ISSN 2045-2322, Vol. 16, article id 13133Article in journal (Refereed) Published
Abstract [en]

The potential of Komagataeibacter (K.) sucrofermentans to valorize polyethylene terephthalate (PET) monomers - ethylene glycol (EG) & disodium terephthalate (Na2-TPA) - and glucose into bacterial nanocellulose (BNC) membranes was investigated using Raman and Fourier transform infrared (FT-IR) spectroscopy. Gravimetric analysis indicated higher BNC yields for K. sucrofermentans nurtured with EG (2.54 g L− 1) compared to pure glucose (2.00 g L− 1) and TPA (1.54 g L− 1). Scanning Electron Microscopy (SEM) images showed that BNC derived from glucose had a dense interconnected fibrillar matrix with uniform porosity, whereas EG and TPA displayed a network structure with less uniform fiber packing. The Segal approach from X-ray diffraction (XRD) showed a lower crystallinity for EG (CrI 62.3%) and substantially lower for TPA (CrI of 18%) than for the commonly known values of glucose (CrI around 70%). Raman spectra of BNC showed that bands between 1050 cm− 1 and 1150 cm− 1 were shifted towards lower wavenumbers compared to AVICEL cellulose, indicating an increased strain on the glycosidic linkages. The Raman spectrum of BNC with TPA as a feedstock showed additional bands at 629, 861, 1125, and 1425 cm− 1 indicating that not all TPA was valorized. FT-IR spectra of the BNC with EG and TPA as feedstocks showed minor transmission and bands at 447, 560, 687, 730, 862, plus between 1500 and 1600 cm− 1 that are typically assigned to PET monomers. The absorption ratio A ~ 1430/ A ~ 897, a measure of crystallinity, gave the values glucose = 1.02, TPA = 0.63, EG = 0.53 showing that glucose as a feedstock produced the highest BNC crystallinity. In the case of glucose and EG, well-structured cellulose membranes were produced, whereas for TPA a composite membrane was produced, paving the way to use PET monomers as a feedstock for biodegradable BNC production.

Place, publisher, year, edition, pages
Springer Nature, 2026
Keywords
Bacterial nanocellulose, Carbon sources, Ethylene glycol (EG), Disodium terephthalate (Na2 TPA), Raman spectroscopy, Fourier Transform Infrared (FTIR) spectroscopy
National Category
Atom and Molecular Physics and Optics
Research subject
Experimental Mechanics; Biochemical Process Engineering
Identifiers
urn:nbn:se:ltu:diva-112184 (URN)10.1038/s41598-026-46886-z (DOI)001747022600008 ()42020601 (PubMedID)2-s2.0-105036797104 (Scopus ID)
Funder
Bio4Energy, 2024–2025, B4E3-TM-3-09
Note

Full text license: CC BY 4.0

Available from: 2025-03-31 Created: 2025-03-31 Last updated: 2026-05-12Bibliographically approved
2. Investigation of the Spatial Generation of Stimulated Raman Scattering Using Computer Simulation and Experimentation
Open this publication in new window or tab >>Investigation of the Spatial Generation of Stimulated Raman Scattering Using Computer Simulation and Experimentation
2022 (English)In: Applied Spectroscopy, ISSN 0003-7028, E-ISSN 1943-3530, Vol. 76, no 11, p. 1307-1316Article in journal (Refereed) Published
Abstract [en]

Stimulated Raman scattering is a phenomenon with potential use in providing real-time molecular information in three-dimensions (3D) of a sample using imaging. For precise imaging, the knowledge about the spatial generation of stimulated Raman scattering is essential. To investigate the spatial behavior in an idealized case, computer simulations and experiments were performed. For the computer simulations, diffraction theory was used for the beam propagation complemented with nonlinear phase modulation describing the interaction between the light and matter. For the experiments, a volume of ethanol was illuminated by an expanded light beam and a plane inside the volume was imaged in transmission. For generating stimulated Raman scattering, a pump beam was focused into this volume and led to a beam dump after passing the volume. The pulse duration of the two beams were 6 ns and the pump beam energy ranged from 1 to 27 mJ. The effect of increasing pump power on the spatial distribution of the Raman gain and the spatial growth of the signal at different interaction lengths between the beam and the sample was investigated. The spatial width of the region where the stimulated Raman scattering signal was generated for experiments and simulation was 0.21 and 0.09 mm, respectively. The experimental and simulation results showed that most of the stimulated Raman scattering is generated close to the pump beam focus and the maximum peak of the Stokes intensity spatially comes shortly after the peak of the pump intensity.

Place, publisher, year, edition, pages
Sage Publications, 2022
Keywords
Stimulated Raman scattering, simulations, experiments, spatial distribution, spatial rate
National Category
Atom and Molecular Physics and Optics Applied Mechanics
Research subject
Experimental Mechanics
Identifiers
urn:nbn:se:ltu:diva-90344 (URN)10.1177/00037028221123593 (DOI)000872145200003 ()36281542 (PubMedID)2-s2.0-85140561101 (Scopus ID)
Funder
Luleå University of TechnologyThe Kempe FoundationsSwedish Foundation for Strategic Research, ITM17-0056
Note

Validerad;2022;Nivå 2;2022-11-03 (sofila)

Available from: 2022-04-21 Created: 2022-04-21 Last updated: 2025-10-21Bibliographically approved
3. Two camera phase contrast imaging using digital correlation imaging of speckles
Open this publication in new window or tab >>Two camera phase contrast imaging using digital correlation imaging of speckles
2024 (English)In: Applied Optics, ISSN 1559-128X, E-ISSN 2155-3165, Vol. 63, no 36, p. 9254-9260Article in journal (Refereed) Published
Place, publisher, year, edition, pages
Optica Publishing Group, 2024
National Category
Applied Mechanics
Research subject
Experimental Mechanics
Identifiers
urn:nbn:se:ltu:diva-111093 (URN)10.1364/ao.540283 (DOI)001379778300006 ()2-s2.0-85212764229 (Scopus ID)
Funder
Swedish Foundation for Strategic Research, ITM17-0056The Kempe Foundations
Note

Validerad;2025;Nivå 2;2025-03-20 (u2);

Available from: 2024-12-17 Created: 2024-12-17 Last updated: 2025-10-21Bibliographically approved
4. Reliability of the image formation in the phase-contrast speckle correlation imaging technique
Open this publication in new window or tab >>Reliability of the image formation in the phase-contrast speckle correlation imaging technique
2025 (English)In: Applied Optics, ISSN 1559-128X, Vol. 64, no 15, p. 4235-4240Article in journal (Refereed) Published
Place, publisher, year, edition, pages
Optica Publishing Group, 2025
National Category
Atom and Molecular Physics and Optics
Research subject
Experimental Mechanics
Identifiers
urn:nbn:se:ltu:diva-112185 (URN)10.1364/AO.561039 (DOI)001504644300011 ()40793134 (PubMedID)2-s2.0-105005504728 (Scopus ID)
Funder
Swedish Foundation for Strategic Research, ITM17-0056The Kempe Foundations
Note

Validerad;2025;Nivå 2;2025-05-26 (u2);

This article has previously appeared as a manuscript in a thesis.

Available from: 2025-03-31 Created: 2025-03-31 Last updated: 2026-02-12Bibliographically approved
5. 3D spatial control and the spatial generation of stimulated Raman scattering in ethanol
Open this publication in new window or tab >>3D spatial control and the spatial generation of stimulated Raman scattering in ethanol
2022 (English)Conference paper, Oral presentation only (Other academic)
Keywords
Stimulated Raman scattering
National Category
Atom and Molecular Physics and Optics
Research subject
Experimental Mechanics
Identifiers
urn:nbn:se:ltu:diva-97135 (URN)
Conference
ECONOS European Conference on Non-linear Optical Spectroscopy, September 25-28, 2022, Kiruna, Sweden
Available from: 2023-05-12 Created: 2023-05-12 Last updated: 2025-10-21Bibliographically approved

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