Development of a second harmonic generation microscope optimized for biomaterial studies
-
Imee Su Martinez
,
Allen Marbert S. Lee
Abstract
A Second harmonic generation (SHG) microscope optimized for biomaterials was developed. The set-up has a non-collinear counter propagating geometry, that allows for the polarization tuning of incoming and outgoing beams. The developed microscope has a resolution of 1.1 µm/line pair. It makes use of a highly sensitive and high-resolution CCD camera that allows for the imaging of delicate biomaterial samples in short acquisition times without burning. This is the first and only functional SHG microscope set-up in the Philippines.
Funding source: Philippine Council for Industry, Energy, and Emerging Technology Research and Development
Funding source: Philippine Council for Agriculture, Aquatic and Natural Resources Research and Development
Award Identifier / Grant number: CHE-18-1-05
Funding source: Industrial Technology Development Institute
Funding source: Department of Science and Technology-Accelerated Science and Technology Human Resource Development Program
Funding source: University of the Philippines Diliman Office of the Vice Chancellor for Research and Development Outright Research Grant
Award Identifier / Grant number: ORG 232323
Acknowledgments
The authors would like to acknowledge funding provided by the Natural Sciences Research Institute (CHE-18-1-05), the Department of Science and Technology (DOST)–PCAARRD, DOST-PCIEERD, the DOST–Industrial Technology Development Institute, the DOST-Accelerated Science and Technology Human Resource Development Programꟷfor personnel support, and the OVCRD Outright Research Grant 232323. We would also like to acknowledge the generosity of Dr. Ricky Nellas for lending his upright microscope, and Dr. Maria Antonette Juinio-Meñez for providing the samples from the Marine Science Institute Bolinao Marine Laboratory.
-
Research ethics: Not applicable.
-
Informed consent: Not applicable.
-
Author contributions: All authors have accepted responsibility for the entire content of this manuscript and approved its submission. Imee Su Martinez is the principal investigator of the study, and wrote the manuscript. Allen Marbert Lee built the first prototype of the SHG microscope in laboratory. Kent Andew Llamson optimized the microscope for biomaterials and improved the microscope by equipping it for future polarization mapping and chirality studies.
-
Use of Large Language Models, AI and Machine Learning Tools: None declared.
-
Conflict of interest: The authors state no conflict of interest.
-
Research funding: Funding was provided by the Natural Sciences Research Institute (CHE-18-1-05), the Department of Science and Technology (DOST)–PCAARRD, DOST-PCIEERD, the DOST–Industrial Technology Development Institute, the DOST-Accelerated Science and Technology Human Resource Development Programꟷfor personnel support, and the OVCRD Outright Research Grant 232323.
-
Data availability: Not applicable.
-
Supporting Information: A video of dry collagen film second harmonic generation, as it was exposed to the atmosphere.
References
1. Franken, P. A.; Hill, A. E.; Peters, C. W.; Weinreich, G. Generation of Optical Harmonics. Phys. Rev. Lett. 1961, 7, 118–20; https://doi.org/10.1103/physrevlett.7.118.Suche in Google Scholar
2. Schawlow, A. L.; Townes, C. H. Infrared and Optical Masers. Phys. Rev. 1958, 112, 1940–9; https://doi.org/10.1103/physrev.112.1940.Suche in Google Scholar
3. Freund, I.; Deutsch, M.; Sprecher, A. Connective Tissue Polarity. Biophys. J. 1986, 50, 693–712; https://doi.org/10.1016/s0006-3495(86)83510-x.Suche in Google Scholar
4. Flörsheimer, M. Second-harmonic Microscopy – New Tool for the Remote Sensing of Interfaces. Phys. Stat. Sol. A 1999, 173, 15–27; https://doi.org/10.1002/(sici)1521-396x(199905)173:1<15::aid-pssa15>3.0.co;2-8.10.1002/(SICI)1521-396X(199905)173:1<15::AID-PSSA15>3.0.CO;2-8Suche in Google Scholar
5. Peterson, M. D.; Hayes, P. L.; Martinez, I. S.; Cass, L. C.; Achtyl, J. L.; Weiss, E. A.; Geiger, F. M. Second Harmonic Generation Imaging with a KHz Amplifier. Opt. Mater. Express 2011, 1, 57–66; https://doi.org/10.1364/ome.1.000057.Suche in Google Scholar
6. Kriech, M. A.; Conboy, J. C. Counterpropagating Second-harmonic Generation: A New Technique for the Invetigation of Molecular Chirality at the Surfaces. J. Opt. Soc. Am. B 2004, 21, 1013–22; https://doi.org/10.1364/josab.21.001013.Suche in Google Scholar
7. Kriech, M. A.; Conboy, J. C. Imaging Chirality with Surface Second Harmonic Generation Spectroscopy. J. Am. Chem. Soc. 2005, 127, 2834–5; https://doi.org/10.1021/ja0430649.Suche in Google Scholar
8. Aghigh, A.; Bancelin, S.; Rivard, M.; Pinsard, M.; Ibrahim, H.; Légaré, F. Second Harmonic Generation Microscopy: A Powerful Tool for Bio-imaging. Biophys. Rev. 2023, 15, 43–70; https://doi.org/10.1007/s12551-022-01041-6.Suche in Google Scholar
9. Hristu, R.; Stanciu, S. G.; Dumitru, A.; Lucian, G.; Eftimie, L. G.; Paun, B.; Tranca, D. E.; Gheorghita, P.; Costache, M.; Stanciu, G. A. PSHG-tISS: A Collection of Polarization-resolved Second Harmonic Generation Microscopy Images of Fixed Tissues. Sci. Data 2022, 9, 1–12; https://doi.org/10.1038/s41597-022-01477-1.Suche in Google Scholar
10. Karunendiran, A.; Mirsanaye, K.; Stewart, B. A.; Barzda, V. Second Harmonic Generation Properties in Chiral Sarcomeres of Drosophila Larval Muscles. Front. Phys. 2022, 10, 1–8; https://doi.org/10.3389/fphy.2022.758709.Suche in Google Scholar
11. Mirsanaye, K.; Castaño, L. U.; Kamaliddin, Y.; Golaraei, A.; Augulis, R.; Kontenis, L.; Done, S.J.; Žurauskas, E.; Stambolic, V.; Wilson, B. C.; Barzda, V. Machine Learning-enabled Cancer Diagnostics with Widefield Polarimetric Second-harmonic Generation Microscopy. Sci. Rep. 2022, 12, 1–14; https://doi.org/10.1038/s41598-022-13623-1.Suche in Google Scholar PubMed PubMed Central
12. Johnson, P. B.; Karvounis, A.; Singh, H. J.; Brereton, C. J.; Bourdakos, K. N.; Lunn, K.; Roberts, J. J. W.; Davies, D. E.; Muskens, O. L.; Jones, M. G.; Sumeet, M. Super-resolved Polarization-enhanced Second-harmonic Generation for Direct Imaging of Nanoscale Changes in Collagen Architecture. Optica 2021, 8, 674–85; https://doi.org/10.1364/optica.411325.Suche in Google Scholar
13. Zhang, C.; Lin, F.; Zhang, Y.; Yang, H.; Lin, D.; He, J.; Liao, C.; Weng, X.; Liu, L.; Wang, Y.; Yu, B.; Qu, J. Super-Resolution Second-Harmonic Generation Imaging with Multifocal Structured Illumination Microscopy. Nano Lett. 2023, 23, 7975–82; https://doi.org/10.1021/acs.nanolett.3c01903.Suche in Google Scholar PubMed
14. Shen, Y. R. Surface Properties Probed by Second-harmonic and Sum-frequency Generation. Nature 1989, 337, 519–25; https://doi.org/10.1038/337519a0.Suche in Google Scholar
15. Boyd, R. Nonlinear Optics; Academic Press: San Diego, 2003.Suche in Google Scholar
16. Lambert, A. G.; Davies, P. B.; Neivandt, D. J. Implementing the Theory of Sum Frequency Generation Vibrational Spectroscopy: A Tutorial Review. Appl. Spec. Rev. 2005, 40, 103–45; https://doi.org/10.1081/asr-200038326.Suche in Google Scholar
17. Sipe, J.; Moss, D.; van Driel, H. Phenomenological Theory of Optical Second- and Third-harmonic Generation from Cubic Centrosymmetric Crystals. Phys. Rev. B 1987, 35, 1129–41; https://doi.org/10.1103/physrevb.35.1129.Suche in Google Scholar PubMed
18. Xia, J.; Wei, Z.; Zhang, J. Demonstration of High Conversion Efficiency to Second Harmonic in a Wide Tuning Range. Opt. Laser Tech. 2000, 32, 241–4; https://doi.org/10.1016/s0030-3992(00)00050-5.Suche in Google Scholar
19. Jeon, Y.; Min, H.; Kim, D.; Oh-e, M. Determination of the Crystalline x-Axis of Quartz by Second-Harmonic Phase Measurement. J. Kor. Phys. Soc. 2005, 46, S159–S62.Suche in Google Scholar
20. Vallejo Ramirez, P. P.; Zammit, J.; Vanderpoorten, O.; Riche, F.; Blé, F-X.; Zhou, X-H.; Spiridon, B.; Valentine, C.; Spasov, S. E.; Oluwasanya, P. W.; Goodfellow, G.; Fantham, M. J.; Siddiqui, O.; Alimagham, F.; Robbins, M.; Stretton, A.; Simatos, D.; Hadeler, O.; Rees, E. J.; Ströhl, F.; Laine, R. F.; Kaminski, C. F. Open-source Optical Projection Tomography of Large Organ Samples. Sci. Rep. 2019, 9, 1–9; https://doi.org/10.1038/s41598-019-52065-0.Suche in Google Scholar PubMed PubMed Central
21. Schneider, C. A.; Rasband, W. S.; Eliceiri, K. W. NIH Image to ImageJ: 25 Years of Image Analysis. Nat. Methods 2012, 9, 671–5; https://doi.org/10.1038/nmeth.2089.Suche in Google Scholar PubMed PubMed Central
22. Sisican, K. M. D.; Torreno, V. P. M.; Yu, E. T.; Conato, M. T. Physicochemical and Biochemical Characterization of Collagen from Stichopus cf. Horrens Tissues for Use as Stimuli-Responsive Thin Films. ACS Omega 2023, 8, 35791–9; https://doi.org/10.1021/acsomega.3c03299.Suche in Google Scholar PubMed PubMed Central
23. Saito, M.; Kunisaki, N.; Urano, N.; Kimura, S. Collagen as the Major Edible Component of Sea Cucumber (Stichopus japonicus). J. Food Sci. 2006, 67, 1319–22; https://doi.org/10.1111/j.1365-2621.2002.tb10281.x.Suche in Google Scholar
24. Campagnola, P. Second Harmonic Generation Imaging Microscopy: Applications to Diseases Diagnostics. Anal. Chem. 2011, 83, 3224–32; https://doi.org/10.1021/ac1032325.Suche in Google Scholar PubMed PubMed Central
25. Bonneel, M.; Hennebert, E.; Arankoc, A. S.; Hwang, D. S.; Lefevre, M.; Pommier, V.; Wattiez, R.; Delroisse, J.; Flammang, P. Molecular Mechanisms Mediating Stiffening in the Mechanically Adaptable Connective Tissues of Sea Cucumbers. Matrix Bio 2022, 108, 39–54; https://doi.org/10.1016/j.matbio.2022.02.006.Suche in Google Scholar PubMed
Supplementary Material
This article contains supplementary material (https://doi.org/10.1515/pac-2024-0383).
© 2025 IUPAC & De Gruyter
Artikel in diesem Heft
- Frontmatter
- In this issue
- Research Articles
- Efficient degradation of 1,2-dichlorobenzene using heterogeneous catalytic ozonation over metal loaded gamma alumina catalysts
- Effect of chemical modification using glyoxylic acid on the stability of α-amylase from Aspergillus fumigatus
- A new, innovative, simple method to determine the concentration of phosphate and sulphate ions in an aqueous extract of plants using conductometric titration
- Metalloporphyrin-mediated oxidative degradation of risperidone under mild conditions: an LC-MS/MS study
- Synthesis of novel ternary herbicide-layered double hydroxide hybrids via the ion exchange method
- Water Quality Index and the quality of freshwater resource uMhlathuze river, Kwazulu-Natal, South Africa: A Review
- Experimental ‘in-Vitro’ investigation on bio-chemical constituents, radical scavenging activity, and reducing power assay of cow urine
- Enhanced biofilm disruption in ESKAPE pathogens through synergistic activity of EPS degrading enzymes
- Green synthesis of silver nanoparticles using a bioflocculant produced by a kombucha tea yeast isolate for antimicrobial and biosafety testing
- Characterization of metabolite compounds from endophytic fungi associated with white turi plant (Sesbania grandiflora) and their antibacterial activity
- Enhanced photocatalytic degradation of methylene blue dye using TiO2 nanoparticles obtained via chemical and green synthesis: a comparative analysis
- Characterization of electrocatalysts for the oxygen evolution reaction OER: a bi-metal study IrM oxides (Ru, and Au)
- Development of a second harmonic generation microscope optimized for biomaterial studies
Artikel in diesem Heft
- Frontmatter
- In this issue
- Research Articles
- Efficient degradation of 1,2-dichlorobenzene using heterogeneous catalytic ozonation over metal loaded gamma alumina catalysts
- Effect of chemical modification using glyoxylic acid on the stability of α-amylase from Aspergillus fumigatus
- A new, innovative, simple method to determine the concentration of phosphate and sulphate ions in an aqueous extract of plants using conductometric titration
- Metalloporphyrin-mediated oxidative degradation of risperidone under mild conditions: an LC-MS/MS study
- Synthesis of novel ternary herbicide-layered double hydroxide hybrids via the ion exchange method
- Water Quality Index and the quality of freshwater resource uMhlathuze river, Kwazulu-Natal, South Africa: A Review
- Experimental ‘in-Vitro’ investigation on bio-chemical constituents, radical scavenging activity, and reducing power assay of cow urine
- Enhanced biofilm disruption in ESKAPE pathogens through synergistic activity of EPS degrading enzymes
- Green synthesis of silver nanoparticles using a bioflocculant produced by a kombucha tea yeast isolate for antimicrobial and biosafety testing
- Characterization of metabolite compounds from endophytic fungi associated with white turi plant (Sesbania grandiflora) and their antibacterial activity
- Enhanced photocatalytic degradation of methylene blue dye using TiO2 nanoparticles obtained via chemical and green synthesis: a comparative analysis
- Characterization of electrocatalysts for the oxygen evolution reaction OER: a bi-metal study IrM oxides (Ru, and Au)
- Development of a second harmonic generation microscope optimized for biomaterial studies
