Development of Two Photon Confocal Laser Scanning System

So, Peter T.

Abstract

The aim of this project is to construct a two-photon confocal microscope with optical ablation and trapping capabilities. The ability to non-invasively monitor cellular systems before and after optical manipulation is crucial. This project will proceed in two stages. First, a two-photon microscope will be construct in LAMMP, then laser tweezers and scissors will be implemented in the new microscope. We are now working toward the completion the first stage. We have purchased a Titanium-Sapphire laser. This is the most important element in the two-photon system because the high peak power femtosecond laser ensures efficient two-photon excitation. We are also in the process of evaluating the effect of two-photon micro-irradiation on biological system. Significant progress has been made in the characterization of two-photon irradiation damage using Chinese hamster ovary (CHO) cells. We found that two-photon irradiation above a threshold dosage can cause auto-fluorescence increase in CHO cell indicating cellular damage. The observed auto-fluorescence increase is providing valuable clues in understanding the mechanism of two-photon damage and will be further studied in the near future.

Funding Agency

• Agency: National Institute of Health (NIH)
• Institute: National Center for Research Resources (NCRR)
• Type: Biotechnology Resource Grants (P41)
• Project #: 5P41RR001192-17
• Application #: 5222590
• Support Year: 17
• Fiscal Year: 1996

Publications

Paugh, Jerry R; Alfonso-Garcia, Alba; Nguyen, Andrew Loc et al. (2018) Characterization of expressed human meibum using hyperspectral stimulated Raman scattering microscopy. Ocul Surf :

Verdel, Nina; Lentsch, Griffin; Balu, Mihaela et al. (2018) Noninvasive assessment of skin structure by combined photothermal radiometry and optical spectroscopy: coregistration with multiphoton microscopy. Appl Opt 57:D117-D122

Friedman, Jacob E; Dobrinskikh, Evgenia; Alfonso-Garcia, Alba et al. (2018) Pyrroloquinoline quinone prevents developmental programming of microbial dysbiosis and macrophage polarization to attenuate liver fibrosis in offspring of obese mice. Hepatol Commun 2:313-328

Kennedy, Gordon T; Lentsch, Griffin R; Trieu, Brandon et al. (2017) Solid tissue simulating phantoms having absorption at 970 nm for diffuse optics. J Biomed Opt 22:76013

Takesh, Thair; Sargsyan, Anik; Lee, Matthew et al. (2017) Evaluating the Whitening and Microstructural Effects of a Novel Whitening Strip on Porcelain and Composite Dental Materials. Dentistry (Sunnyvale) 7:

Jonscher, Karen R; Stewart, Michael S; Alfonso-Garcia, Alba et al. (2017) Early PQQ supplementation has persistent long-term protective effects on developmental programming of hepatic lipotoxicity and inflammation in obese mice. FASEB J 31:1434-1448

Alfonso-García, Alba; Paugh, Jerry; Farid, Marjan et al. (2017) A machine learning framework to analyze hyperspectral stimulated Raman scattering microscopy images of expressed human meibum. J Raman Spectrosc 48:803-812

Takesh, Thair; Sargsyan, Anik; Anbarani, Afarin et al. (2017) Effects of a Novel Whitening Formulation on Dental Enamel. Dentistry (Sunnyvale) 7:

Malacrida, Leonel; Astrada, Soledad; Briva, Arturo et al. (2016) Spectral phasor analysis of LAURDAN fluorescence in live A549 lung cells to study the hydration and time evolution of intracellular lamellar body-like structures. Biochim Biophys Acta 1858:2625-2635

Choi, Bernard; Tan, Wenbin; Jia, Wangcun et al. (2016) The Role of Laser Speckle Imaging in Port-Wine Stain Research: Recent Advances and Opportunities. IEEE J Sel Top Quantum Electron 2016: