Low Efficiency Upconversion Nanoparticles for High-Resolution Coalignment of Near-Infrared and Visible Light Paths on a Light Microscope
| Autor: | Andrew C. Kummel, Julie C. Canman, Adrian Garcia Badaracco, Sophia M. Hirsch, Mimi Shirasu-Hiza, Tim Davies, Jun Hong Park, Julien Dumont, Sriramkumar Sundaramoorthy |
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| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Photon Materials science business.industry Near-infrared spectroscopy High resolution 02 engineering and technology 021001 nanoscience & nanotechnology Article law.invention 03 medical and health sciences 030104 developmental biology Optics Optical microscope law Excited state Microscopy Optoelectronics General Materials Science 0210 nano-technology business Luminescence Visible spectrum |
| Zdroj: | ACS Applied Materials & Interfaces. 9:7929-7940 |
| ISSN: | 1944-8252 1944-8244 |
| DOI: | 10.1021/acsami.6b15322 |
| Popis: | The combination of near infrared (NIR) and visible wavelengths in light microscopy for biological studies is increasingly common. For example, many fields of biology are developing the use of NIR for optogenetics, in which an NIR laser induces a change in gene expression and/or protein function. One major technical barrier in working with both NIR and visible light on an optical microscope is obtaining their precise coalignment at the imaging plane position. Photon upconverting particles (UCPs) can bridge this gap as they are excited by NIR light but emit in the visible range via an anti-Stokes luminescence mechanism. Here, two different UCPs have been identified, high-efficiency micro540-UCPs and lower efficiency nano545-UCPs, that respond to NIR light and emit visible light with high photostability even at very high NIR power densities (>25,000 Suns). Both of these UCPs can be rapidly and reversibly excited by visible and NIR light and emit light at visible wavelengths detectable with standard emission settings used for Green Fluorescent Protein (GFP), a commonly used genetically-encoded fluorophore. However, the high efficiency micro540-UCPs were suboptimal for NIR and visible light coalignment, due to their larger size and spatial broadening from particle-to-particle energy transfer consistent with a long lived excited state and saturated power dependence. In contrast, the lower efficiency nano-UCPs were superior for precise coalignment of the NIR beam with the visible light path (~2 µm versus ~8 µm beam broadening respectively) consistent with limited particle-to-particle energy transfer, superlinear power dependence for emission, and much smaller particle size. Furthermore, the nano-UCPs were superior to a traditional two-camera method for NIR and visible light path alignment in an in vivo Infrared-Laser-Evoked Gene Operator (IR-LEGO) optogenetics assay in the budding yeast S. cerevisiae. In summary, nano-UCPs are powerful new tools for coaligning NIR and visible light paths on a light microscope. |
| Databáze: | OpenAIRE |
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