High-contrast single-particle tracking by selective focal plane illumination microscopy

doi:10.1364/OE.16.007142

High-contrast single-particle tracking by selective focal plane illumination microscopy

Jörg G. Ritter, Roman Veith, Jan-Peter Siebrasse, and Ulrich Kubitscheck »View Author Affiliations

Optics Express, Vol. 16, Issue 10, pp. 7142-7152 (2008)
http://dx.doi.org/10.1364/OE.16.007142

View Full Text Article

Enhanced HTML Acrobat PDF (2235 KB)

Journal Search
Article Lookup

Article Tools

Citations

Alert me when this article is cited
Export Citation/Save

Abstract
Article Info
References (17)
Cited By
Figures (6)
Metrics
Related Content

Abstract

Wide-field single molecule microscopy is a versatile tool for analyzing dynamics and molecular interactions in biological systems. In extended three-dimensional systems, however, the method suffers from intrinsic out-of-focus fluorescence. We constructed a high-resolution selective plane illumination microscope (SPIM) to efficiently solve this problem. The instrument is an optical sectioning microscope featuring the high speed and high sensitivity of a video microscope. We present theoretical calculations and quantitative measurements of the illumination light sheet thickness yielding 1.7 µm (FWHM) at 543 nm, 2.0 µm at 633 nm, and a FWHM of the axial point spread function of 1.13 µm. A direct comparison of selective plane and epi-illumination of model samples with intrinsic background fluorescence illustrated the clear advantage of SPIM for such samples. Single fluorescent quantum dots in aqueous solution are readily visualized and tracked proving the suitability of our setup for the study of fast and dynamic processes in spatially extended biological specimens.

OCIS Codes
(170.3880) Medical optics and biotechnology : Medical and biological imaging
(170.6920) Medical optics and biotechnology : Time-resolved imaging
(180.0180) Microscopy : Microscopy
(180.2520) Microscopy : Fluorescence microscopy
(180.6900) Microscopy : Three-dimensional microscopy

ToC Category:
Microscopy

History
Original Manuscript: January 30, 2008
Revised Manuscript: March 7, 2008
Manuscript Accepted: April 24, 2008
Published: May 2, 2008

Virtual Issues
Vol. 3, Iss. 6 Virtual Journal for Biomedical Optics

Citation
Jörg G. Ritter, Roman Veith, Jan-Peter Siebrasse, and Ulrich Kubitscheck, "High-contrast single-particle tracking by selective focal plane illumination microscopy," Opt. Express 16, 7142-7152 (2008)
http://www.opticsinfobase.org/oe/abstract.cfm?URI=oe-16-10-7142

Sort: Author | Year | Journal | Reset

References

J. P. Siebrasse, D. Grunwald, and U. Kubitscheck, "Single-molecule tracking in eukaryotic cell nuclei," Anal. Bioanal. Chem. (2006). [CrossRef] [PubMed]
H. Siedentopf and R. Zsigmondy, "�?ber Sichtbarmachung und Grössenbestimmung ultramikroskopischer Teilchen," Annalen der Physik 4, 1-39 (1903).
A. H. Voie, D. H. Burns, and F. A. Spelman, "Orthogonal-Plane Fluorescence Optical Sectioning - 3-Dimensional Imaging of Macroscopic Biological Specimens," J. Microsc. 170, 229-236 (1993). [CrossRef] [PubMed]
J. Huisken, J. Swoger, F. Del Bene, J. Wittbrodt, and E. H. Stelzer, "Optical sectioning deep inside live embryos by selective plane illumination microscopy," Science 305, 1007-1009 (2004). [CrossRef] [PubMed]
J. Huisken and D. Y. Stainier, "Even fluorescence excitation by multidirectional selective plane illumination microscopy (mSPIM)," Opt. Lett. 32, 2608-2610 (2007). [CrossRef] [PubMed]
J. A. Buytaert and J. J. Dirckx, "Design and quantitative resolution measurements of an optical virtual sectioning three-dimensional imaging technique for biomedical specimens, featuring two-micrometer slicing resolution," J Biomed. Opt. 12, 014039 (2007). [CrossRef] [PubMed]
H. U. Dodt, U. Leischner, A. Schierloh, N. Jahrling, C. P. Mauch, K. Deininger, J. M. Deussing, M. Eder, W. Zieglgansberger, and K. Becker, "Ultramicroscopy: three-dimensional visualization of neuronal networks in the whole mouse brain," Nat. Methods 4, 331-336 (2007). [CrossRef] [PubMed]
W. Alt, "An objective lens for efficient fluorescence detection of single atoms," Optik 113, 142-144 (2002). [CrossRef]
B. E. A. Saleh and M. C. Teich, Fundamentals of Photonics (Wiley, 2007).
C. J. Engelbrecht and E. H. Stelzer, "Resolution enhancement in a light-sheet-based microscope (SPIM)," Opt. Lett. 31, 1477-1479 (2006). [CrossRef] [PubMed]
J. H. E. Peter W. Millonni, Lasers (Wiley & Sons, 1989).
M. Born and E. Wolf, Principles of Optics (Cambridge University Press, Cambridge, 1997).
D. Wright, P. Greve, J. Fleischer, and L. Austin, "Laser beam width, divergence and beam propagation factor - an international standardization approach," Opt. Quantum Electron. 24, S993-S1000 (1992). [CrossRef]
T. Soop, B. Ivarsson, B. Bjorkroth, N. Fomproix, S. Masich, V. C. Cordes, and B. Daneholt, "Nup153 affects entry of messenger and ribosomal ribonucleoproteins into the nuclear basket during export," Mol. Biol. Cell 16, 5610-5620 (2005). [CrossRef] [PubMed]
M. Abramowitz, P. J. Magelhaes, and S. J. Ram, "Image Processing with ImageJ," Biophot. Int. 11, 36-42 (2004).
T. Kues, A. Dickmanns, R. Lührmann, R. Peters, and U. Kubitscheck, "High intranuclear mobility and dynamic clustering of the splicing factor U1 snRNP observed by single particle tracking," Proc. Natl. Acad. Sci. U S A 98, 12021-12026 (2001). [CrossRef] [PubMed]
D. Grünwald, A. Hoekstra, T. Dange, V. Buschmann, and U. Kubitscheck, "Direct observation of single protein molecules in aqueous solution," ChemPhysChem 7, 812-815 (2006). [CrossRef] [PubMed]

Abramowitz, M.
Alt, W.
Austin, L.
Becker, K.
Bjorkroth, B.
Burns, D. H.
Buschmann, V.
Buytaert, J. A.
Cordes, V. C.
Daneholt, B.
Dange, T.
Deininger, K.
Del Bene, F.
Deussing, J. M.
Dickmanns, A.
Dirckx, J. J.
Dodt, H. U.
Eder, M.
Engelbrecht, C. J.
Fleischer, J.
Fomproix, N.
Greve, P.
Grünwald, D.
Hoekstra, A.
Huisken, J.
Ivarsson, B.
Jahrling, N.
Kubitscheck, U.
Kues, T.
Leischner, U.
Lührmann, R.
Magelhaes, P. J.
Masich, S.
Mauch, C. P.
Peters, R.
Ram, S. J.
Schierloh, A.
Siedentopf, H.
Soop, T.
Spelman, F. A.
Stainier, D. Y.
Stelzer, E. H.
Swoger, J.
Voie, A. H.
Wittbrodt, J.
Wright, D.
Zieglgansberger, W.
Zsigmondy, R.

Annalen der Physik

H. Siedentopf and R. Zsigmondy, "�?ber Sichtbarmachung und Grössenbestimmung ultramikroskopischer Teilchen," Annalen der Physik 4, 1-39 (1903).

Biophot. Int.

M. Abramowitz, P. J. Magelhaes, and S. J. Ram, "Image Processing with ImageJ," Biophot. Int. 11, 36-42 (2004).

ChemPhysChem

D. Grünwald, A. Hoekstra, T. Dange, V. Buschmann, and U. Kubitscheck, "Direct observation of single protein molecules in aqueous solution," ChemPhysChem 7, 812-815 (2006). [CrossRef] [PubMed]

J Biomed. Opt.

J. A. Buytaert and J. J. Dirckx, "Design and quantitative resolution measurements of an optical virtual sectioning three-dimensional imaging technique for biomedical specimens, featuring two-micrometer slicing resolution," J Biomed. Opt. 12, 014039 (2007). [CrossRef] [PubMed]

J. Microsc.

A. H. Voie, D. H. Burns, and F. A. Spelman, "Orthogonal-Plane Fluorescence Optical Sectioning - 3-Dimensional Imaging of Macroscopic Biological Specimens," J. Microsc. 170, 229-236 (1993). [CrossRef] [PubMed]

Mol. Biol. Cell

T. Soop, B. Ivarsson, B. Bjorkroth, N. Fomproix, S. Masich, V. C. Cordes, and B. Daneholt, "Nup153 affects entry of messenger and ribosomal ribonucleoproteins into the nuclear basket during export," Mol. Biol. Cell 16, 5610-5620 (2005). [CrossRef] [PubMed]

Nat. Methods

H. U. Dodt, U. Leischner, A. Schierloh, N. Jahrling, C. P. Mauch, K. Deininger, J. M. Deussing, M. Eder, W. Zieglgansberger, and K. Becker, "Ultramicroscopy: three-dimensional visualization of neuronal networks in the whole mouse brain," Nat. Methods 4, 331-336 (2007). [CrossRef] [PubMed]

Opt. Lett.

Opt. Quantum Electron.

D. Wright, P. Greve, J. Fleischer, and L. Austin, "Laser beam width, divergence and beam propagation factor - an international standardization approach," Opt. Quantum Electron. 24, S993-S1000 (1992). [CrossRef]

Optik

W. Alt, "An objective lens for efficient fluorescence detection of single atoms," Optik 113, 142-144 (2002). [CrossRef]

Proc. Natl. Acad. Sci. U S A

T. Kues, A. Dickmanns, R. Lührmann, R. Peters, and U. Kubitscheck, "High intranuclear mobility and dynamic clustering of the splicing factor U1 snRNP observed by single particle tracking," Proc. Natl. Acad. Sci. U S A 98, 12021-12026 (2001). [CrossRef] [PubMed]

Science

J. Huisken, J. Swoger, F. Del Bene, J. Wittbrodt, and E. H. Stelzer, "Optical sectioning deep inside live embryos by selective plane illumination microscopy," Science 305, 1007-1009 (2004). [CrossRef] [PubMed]

Other

J. P. Siebrasse, D. Grunwald, and U. Kubitscheck, "Single-molecule tracking in eukaryotic cell nuclei," Anal. Bioanal. Chem. (2006). [CrossRef] [PubMed]
B. E. A. Saleh and M. C. Teich, Fundamentals of Photonics (Wiley, 2007).
J. H. E. Peter W. Millonni, Lasers (Wiley & Sons, 1989).
M. Born and E. Wolf, Principles of Optics (Cambridge University Press, Cambridge, 1997).

Cited By

Alert me when this paper is cited

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.

Click here to see a list of articles that cite this paper