Variable immersion microscopy with a high numerical aperture
| Autor: | Ishida, Keita, Naruse, Kanta, Mizouchi, Yuta, Ogawa, Y., Matsushita, Michio, Shimi, Takeshi, Kimura, Hiroshi, FUJIYOSHI, SATORU |
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| Rok vydání: | 2021 |
| Předmět: |
Microscopy
Materials science Microscope business.industry 02 engineering and technology 021001 nanoscience & nanotechnology 01 natural sciences Refraction Atomic and Molecular Physics and Optics law.invention Numerical aperture 010309 optics Lens (optics) Wavelength Refractometry Optics Optical microscope law 0103 physical sciences Immersion 0210 nano-technology business Refractive index |
| Zdroj: | Optics letters. 46(4) |
| ISSN: | 1539-4794 |
| Popis: | Three-dimensional (3D) optical microscopy with a high numerical aperture (NA) remains challenging for thick biological specimens owing to aberrations arising from interface refractions. We developed a variable immersion lens (VIL) to passively minimize these aberrations. A VIL is a high-NA concentric meniscus lens and was used in combination with an aberration-corrected high-NA reflecting objective (TORA-FUJI mirror). Wave-optics simulation at a wavelength of 488 nm showed that a VIL microscope enables diffraction-limited 1.2-NA imaging in water (refractive index of 1.34) at a depth of 0.3 mm by minimizing aberrations due to refraction of a sample interface. Another aberration due to the refractive index mismatching between a mounting medium, and an object can also be corrected by the VIL system, because various fluids with different refractive indices can be used as mounting media for the VIL. As a result of correcting the two aberrations at the same time, we experimentally demonstrated that a 6 µm diameter fluorescent bead can be imaged to the true dimensions in 3D. |
| Databáze: | OpenAIRE |
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