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Laser induced degradation of bacterial spores during micro-Raman spectroscopy
Umeå University, Faculty of Science and Technology, Department of Physics.
Umeå University, Faculty of Science and Technology, Department of Physics.
Umeå University, Faculty of Science and Technology, Department of Physics.
Umeå University, Faculty of Science and Technology, Department of Physics.
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2022 (English)In: Spectrochimica Acta Part A - Molecular and Biomolecular Spectroscopy, ISSN 1386-1425, E-ISSN 1873-3557, Vol. 265, article id 120381Article in journal (Refereed) Published
Abstract [en]

Micro-Raman spectroscopy combined with optical tweezers is a powerful method to analyze how the biochemical composition and molecular structures of individual biological objects change with time. In this work we investigate laser induced effects in the trapped object. Bacillus thuringiensis spores, which are robust organisms known for their resilience to light, heat, and chemicals are used for this study. We trap spores and monitor the Raman peak from CaDPA (calcium dipicolinic acid), which is a chemical protecting the spore core. We see a correlation between the amount of laser power used in the trap and the release of CaDPA from the spore. At a laser power of 5 mW, the CaDPA from spores in water suspension remain intact over the 90 min experiment, however, at higher laser powers an induced effect could be observed. SEM images of laser exposed spores (after loss of CaDPA Raman peak was confirmed) show a notable alteration of the spores' structure. Our Raman data indicates that the median dose exposure to lose the CaDPA peak was ∼60 J at 808 nm. For decontaminated/deactivated spores, i.e., treated in sodium hypochlorite or peracetic acid solutions, the sensitivity on laser power is even more pronounced and different behavior could be observed on spores treated by the two chemicals. Importantly, the observed effect is most likely photochemical since the increase of the spore temperature is in the order of 0.1 K as suggested by our numerical multiphysics model. Our results show that care must be taken when using micro-Raman spectroscopy on biological objects since photoinduced effects may substantially affect the results.

Place, publisher, year, edition, pages
Elsevier, 2022. Vol. 265, article id 120381
Keywords [en]
Bacteria, Decontamination, Multiphysics modelling, Optical tweezers, Raman spectroscopy, Spores
National Category
Biophysics Atom and Molecular Physics and Optics Cell Biology
Identifiers
URN: urn:nbn:se:umu:diva-187284DOI: 10.1016/j.saa.2021.120381ISI: 000709268600014PubMedID: 34562861Scopus ID: 2-s2.0-85115389622OAI: oai:DiVA.org:umu-187284DiVA, id: diva2:1591732
Funder
Swedish Research Council, 2019-04016The Kempe Foundations, JCK-1916.2Swedish Armed Forces, 470-A400821Available from: 2021-09-07 Created: 2021-09-07 Last updated: 2025-09-30Bibliographically approved
In thesis
1. KNOW YOUR ENEMY: Characterizing Pathogenic Biomaterials Using Laser Tweezers
Open this publication in new window or tab >>KNOW YOUR ENEMY: Characterizing Pathogenic Biomaterials Using Laser Tweezers
2022 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Diseases caused by pathogenic agents such as bacteria and viruses result in devastating costs on personal and societal levels. However, it is not just the emergence of new diseases that is problematic. Antibiotic resistance among bacteria makes uncomplicated infections difficult and lethal. Resilient disease-causing spores spread in hospitals, the food industry, and water supplies requiring effective detection and disinfection methods. Further, we face complex neurological diseases where no effective treatment or diagnostic methods exist. Thus, we must increase our fundamental understanding of these diseases to develop effective diagnostic, detection, disinfection, and treatment methods.

Classically, the methods used for detecting and studying the underlying mechanics of pathogenic agents work on a large scale, measuring the average macroscopic behavior and properties of these pathogens. However, just as with humans, the average behavior is not always representative of individual behavior. Therefore, it is also essential to investigate the characteristics of these pathogens on a single cell or particle level. 

This thesis develops and applies optical techniques to characterize pathogenic biomaterial on a single cell or particle level. At the heart of all these studies is our Optical Tweezers (OT) instrument. OT are a tool that allows us to reach into the microscopic world and interact with it. Finally, by combining OT with other experimental techniques, we can chemically characterize biomaterials and develop assays that mimic different biological settings. Using these tools, we investigate bacterial adhesion, disinfection, and detection of pathogenic spores and proteins.

Hopefully, the insights of these studies can lessen the burden on society caused by diseases by helping others develop effective treatment, diagnostic, detection, and disinfection methods in the future. 

Place, publisher, year, edition, pages
Umeå: Umeå universitet, 2022. p. 73
Keywords
Optical Tweezers, Laser Tweezers, Raman Spectroscopy, Bacterial Adhesion, Biophysics, Pili, Bacterial Spores, Endospores, Oocysts, Cryptosporidium, Optics
National Category
Biophysics Atom and Molecular Physics and Optics
Research subject
biology; Physics
Identifiers
urn:nbn:se:umu:diva-192471 (URN)978-91-7855-726-4 (ISBN)978-91-7855-727-1 (ISBN)
Public defence
2022-03-11, NAT.D.410, Naturvetarhuset, Umeå, 09:00 (English)
Opponent
Supervisors
Available from: 2022-02-18 Created: 2022-02-14 Last updated: 2025-02-20Bibliographically approved
2. Spotlight the killer: detecting harmful chemical and biological agents using optical spectroscopy
Open this publication in new window or tab >>Spotlight the killer: detecting harmful chemical and biological agents using optical spectroscopy
2025 (English)Doctoral thesis, comprehensive summary (Other academic)
Alternative title[sv]
Lyset på mördaren : detektion av skadliga kemiska och biologiska ämnen med hjälp av optisk spektroskopi
Abstract [en]

Harmful chemical and biological agents are a significant threat to health and prosperity worldwide. Recent years have seen an increase in wars and conflicts around the globe, raising concerns about the potential deployment of chemical and biological warfare agents. On a less speculative level, harmful chemicals such as narcotic substances cause immense humanitarian and economic damage through overdoses and associated healthcare costs, while microbes such as pathogenic bacteria and parasites cause hospital-acquired infections and food spoilage at a cost of approximately 1 trillion euros every year. To combat the threat of these harmful agents, we must thus develop rapid and effective detection and diagnostic methods for harmful agents, allowing us to effectively deploy specific treatments and preventative measures.

Classically, while there exist numerous methods for the detection of both harmful chemical and biological agents, they often come with limitations that inhibit their effectiveness. These inhibitions often take the form of bulky equipment that is difficult to apply in the field or time-consuming preparation and measurement processes.

In this thesis we will explore an alternative category of assays for detecting and characterizing harmful materials – optical spectroscopy. Optical spectroscopy is a category of material characterization methods that use light to probe a material. While probing the material, we receive a signal characteristic of the molecules, chemical, and biological structure of our material. These optical spectroscopic methods, such as Raman spectroscopy and fluorescence spectroscopy, can be used to characterize a material within the span of minutes or even seconds, making them ideal for detection applications. Furthermore, they can often be made portable or even handheld, making them a great tool for initial field indication of harmful materials, ahead of thorough lab analysis.

I sincerely hope the studies presented herein can serve as a stepping stone to future technologies and detection assays, capable of saving both money and lives. 

Place, publisher, year, edition, pages
Umeå: Umeå University, 2025. p. 72
Keywords
Sensing, Raman spectroscopy, SERS, Fluorescence spectroscopy, CWA, nerve agents, bacterial spores, Cryptosporidium
National Category
Atom and Molecular Physics and Optics
Identifiers
urn:nbn:se:umu:diva-244830 (URN)978-91-8070-780-0 (ISBN)978-91-8070-779-4 (ISBN)
Public defence
2025-10-24, Aula Anatomica, Biologihuset, 907 36, Umeå, Umeå, 13:00 (English)
Opponent
Supervisors
Note

This work was done in collaboration with, and with support from, the Swedish Defece Research Agency (FOI).

Available from: 2025-10-03 Created: 2025-09-30 Last updated: 2025-10-22Bibliographically approved

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Malyshev, DmitryÖberg, RasmusDahlberg, TobiasWiklund, KristerAndersson, Magnus

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Spectrochimica Acta Part A - Molecular and Biomolecular Spectroscopy
BiophysicsAtom and Molecular Physics and OpticsCell Biology

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