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Materials and Methods
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Sheet–Based Fluorescence

Profile image of Herbert SchneckenburgerHerbert Schneckenburger

2016

https://doi.org/10.1117/1.JBO.17.10.101518
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Abstract

Preparation strategy and illumination of three-dimensional cell cultures in light

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References (25)

  1. J. Pawley, Handbook of biological confocal microscopy, Plenum Press, New York (1990).
  2. R. H. Webb, "Confocal optical microscopy," Rep. Prog. Phys. 59(3), 427-471 (1996).
  3. W. Denk, J. H. Strickler, and W. W. Webb, "Two-photon laser scanning fluorescence microscopy," Science 248(4951), 73-76 (1990).
  4. K. König, "Multiphoton microscopy in life sciences," J. Microsc. 200(Pt. 2), 83-104 (2000).
  5. M. A. Neil, R. Juskaitis, and T. Wilson, "Method of obtaining optical sectioning by using structured light in a conventional microscope," Opt. Lett. 22(24), 1905-1907 (1997).
  6. M. G. L. Gustafsson et al., "Three-dimensional resolution doubling in wide-field fluorescence microscopy by structured illumination," Biophys. J. 94(12), 4957-4970 (2008).
  7. M. Wagner et al., "Light dose is a limiting factor to maintain cell via- bility in fluorescence microscopy and single molecule detection," Int. J. Mol. Sci. 11(3), 956-966 (2010).
  8. H. Schneckenburger et al., "Light exposure and cell viability in fluor- escence microscopy," J. Microsc. 245(3), 311-318 (2012).
  9. J. Huisken and D. Y. R. Stainier, "Selective plane illumination microscopy techniques in development biology," Development 136(12), 1963-1975 (2009).
  10. P. A. Santi, "Light sheet fluorescence microscopy: a review," J. Histo- chem. Cytochem. 59(2), 129-138 (2011).
  11. J. G. Ritter et al., "Light sheet microscopy for single molecule tracking in living tissue," PLoS One 5(7), e11639 (2010).
  12. Y. Wu et al., "Inverted selective plane illumination microscopy (iSPIM) enables coupled cell identity lineaging and neurodevelopmental ima- ging in Caenorhabdidits elegans," PNAS 108(43), 17708-17713 (2011).
  13. J. Mertz and J. Kim, "Scanning light-sheet microscopy in the whole mouse brain with HiLo background rejection," J. Biomed. Opt. 15(1), 016027 (2010).
  14. J. Huisken et al., "Optical sectioning deep inside live embryos by SPIM," Science 305(5686), 1007-1009 (2004).
  15. F. O. Fahrbach and A. Rohrbach, "A line-scanned light-sheet micro- scope with phase shaped self-reconstructing beams," Opt. Express 18(23), 24229-24244 (2010).
  16. L. A. Kunz-Schughart et al., "The use of 3-D cultures for high through- put screening: the multi-cellular spheroid model," J. Biomol. Screen. 9(4), 273-285 (2004).
  17. H. Schneckenburger et al., "Multi-dimensional fluorescence micro- scopy of living cells," J. Biophotonics 4(3), 143-149 (2011).
  18. C. Dunsby, "Optically sectioned imaging by oblique plane microscopy," Opt. Express 16(25), 20306-20316 (2008). Journal of Biomedical Optics 101518-4 October 2012 • Vol. 17(10)
  19. Bruns et al.: Preparation strategy and Illumination of three-dimensional cell cultures : : : Downloaded From: https://www.spiedigitallibrary.org/journals/Journal-of-Biomedical-Optics on 13 Jan 2022 Terms of Use: https://www.spiedigitallibrary.org/terms-of-use
  20. K. Greger, J. Swoger, and E. H. K. Stelzer, "Basic building units and properties of a fluorescence single plane illumination microscope," Rev. Sci. Instrum. 78(2), 023705 (2007).
  21. M. Gutscher et al., "Real-time imaging of the intracellular glutathione redox potential," Nat. Methods 5(6), 553-559 (2008).
  22. M. Gutscher et al., "Proximity-based protein thiol oxidation by H 2 O 2 - scavenging peroxidases," J. Biol. Chem. 284(46), 31532-31540 (2009).
  23. B. Angres et al., "A membrane-bound FRET-based caspase sensor for detection of apoptosis using fluorescence lifetime and total internal reflection microscopy," Cytometry 75A(5), 420-427 (2009).
  24. H. Hardelauf et al., "Microarrays for the scalable production of metabolically relevant tumour spheroids: a tool for modulating chemo- sensitivity traits," Lab Chip 11(3), 419-428 (2011). Journal of Biomedical Optics 101518-5 October 2012 • Vol. 17(10)
  25. Bruns et al.: Preparation strategy and Illumination of three-dimensional cell cultures : : : Downloaded From: https://www.spiedigitallibrary.org/journals/Journal-of-Biomedical-Optics on 13 Jan 2022 Terms of Use: https://www.spiedigitallibrary.org/terms-of-use

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