4Pi microscopy of type A with 1-photon excitation in biological fluorescence imaging
Optics Express, Vol. 15, Issue 5, pp. 2459-2467 (2007)
http://dx.doi.org/10.1364/OE.15.002459
Enhanced HTML
Acrobat PDF (570 KB)
Abstract
We demonstrate that oil immersion lenses with a semi-aperture angle ≥ 74° enable 4Pi confocal fluorescence microscopy of type A with 1-photon excitation. The axial sidelobes amount to < 50 % of the main diffraction maximum, implying that lobe induced artifacts can be removed from the image data. The advancement reported herein enables a relative inexpensive implementation of 4Pi microscopy, providing axially superresolved 3D-imaging in transparent samples. As an example, we show dual-color 4Pi images of double stained Golgi stacks in a mammalian cell with 110 nm axial resolution. The resolution can be further enhanced to values slightly below 100 nm by image deconvolution.
© 2007 Optical Society of America
OCIS Codes
(100.6640) Image processing : Superresolution
(180.1790) Microscopy : Confocal microscopy
(180.2520) Microscopy : Fluorescence microscopy
(180.6900) Microscopy : Three-dimensional microscopy
ToC Category:
Microscopy
History
Original Manuscript: January 10, 2007
Manuscript Accepted: February 19, 2007
Published: March 5, 2007
Virtual Issues
Vol. 2, Iss. 4 Virtual Journal for Biomedical Optics
Citation
Marion Lang, Tobias Müller, Johann Engelhardt, and Stefan W. Hell, "4Pi microscopy of type A with 1-photon excitation in biological fluorescence imaging," Opt. Express 15, 2459-2467 (2007)
http://www.opticsinfobase.org/oe/abstract.cfm?URI=oe-15-5-2459
Sort: Year | Journal | Reset
References
- S. Hell and E. H. K. Stelzer, "Properties of a 4Pi-confocal fluorescence microscope", J. Opt. Soc. Am. A 9, 2159-2166 (1992). [CrossRef]
- S. W. Hell, "Double-scanning confocal microscope," European Patent 0491289 (1990).
- M. G. L. Gustafsson et al., "Sevenfold improvement of axial resolution in 3D widefield microscopy using two objective lenses," Proc. SPIE 2412, 147-156 (1995). [CrossRef]
- M. G. L. Gustafsson et al., "I5M: 3D widefield light microscopy with better than 100 nm axial resolution", J. Microsc. 195, 10-16 (1999). [CrossRef] [PubMed]
- S. W. Hell and E. H. K. Stelzer, "Fundamental improvement of resolution with a 4Pi-confocal fluorescence microscope using two-photon excitation," Opt. Commun. 93, 277-282 (1992). [CrossRef]
- S. W. Hell, "Toward fluorescence nanoscopy," Nat. Biotechnol. 21, 1347-1355 (2003). [CrossRef] [PubMed]
- R. Medda et al., "4Pi Microscopy of Quantum Dot-Labeled Cellular Structures," J. Struct. Biol. 156, 517-523 (2006). [CrossRef] [PubMed]
- T. Staudt et al., "2, 2'-Thiodiethanol: A new water soluble mounting medium for high resolution optical microscopy," Microsc. Res. Tech. 70, 1-9 (2007). [CrossRef]
- A. Egner et al., "Fast 100-nm resolution 3D-microscope reveals structural plasticity of mitochondria in live yeast," Proc. Natl. Acad. Sci. USA 99, 3370-3375 (2002). [CrossRef] [PubMed]
- B. Richards and E. Wolf, "Electromagnetic diffraction in optical systems II. Structure of the image field in an aplanatic system," Proc. R. Soc. Lond. A 253, 358-379 (1959). [CrossRef]
- G. Hildenbrand et al., "Nano-sizing of specific gene domains in intact human cell nuclei by spatially modulated illumination light microscopy," Biophys. J. 88, 4312-4318 (2005). [CrossRef] [PubMed]
- H. Gugel et al., "Cooperative 4Pi excitation and detection yields 7-fold sharper optical sections in live cell microscopy," Biophys. J., 87, 4146-4152 (2004). [CrossRef] [PubMed]
- M. Nagorni and S. W. Hell, "Coherent use of opposing lenses for axial resolution increase in fluorescence microscopy. I. Comparative study of concepts," J. Opt. Soc. Am. A 18, 36-48 (2001). [CrossRef]
- W. H. Richardson, "Bayesian-based iterative method of image restoration," J. Opt. Soc. Am. 62, 55-59 (1972). [CrossRef]
- M. Nagorni and S. W. Hell, "Coherent use of opposing lenses for axial resolution increase in fluorescence microscopy. II. Power and limitation of nonlinear image restoration," J. Opt. Soc. Am. A 18, 49-54 (2001). [CrossRef]
Cited By |
OSA is able to provide readers links to articles that cite this paper by participating in CrossRef's Cited-By Linking service. CrossRef includes content from more than 3000 publishers and societies. In addition to listing OSA journal articles that cite this paper, citing articles from other participating publishers will also be listed.





OSA is a member of 