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In vivo fluorescence imaging with high-resolution microlenses

Abstract

Micro-optics are increasingly used for minimally invasive in vivo imaging, in miniaturized microscopes and in lab-on-a-chip devices. Owing to optical aberrations and lower numerical apertures, a main class of microlens, gradient refractive index lenses, has not achieved resolution comparable to conventional microscopy. Here we describe high-resolution microlenses, and illustrate two-photon imaging of dendritic spines on hippocampal neurons and dual-color nonlinear optical imaging of neuromuscular junctions in live mice.

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Figure 1: Aberration-corrected micro-objectives enable high-resolution imaging.
Figure 2: In vivo two-photon imaging with high-resolution micro-objectives permits superior resolution than with uncorrected GRIN lenses and enables visualization of neuronal dendritic spines.

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References

  1. Flusberg, B.A. et al. Nat. Methods 2, 941–950 (2005).

    Article  CAS  Google Scholar 

  2. Levene, M.J., Dombeck, D.A., Kasischke, K.A., Molloy, R.P. & Webb, W.W. J. Neurophysiol. 91, 1908–1912 (2004).

    Article  Google Scholar 

  3. Alencar, H., Mahmood, U., Kawano, Y., Hirata, T. & Weissleder, R. Neoplasia 7, 977–983 (2005).

    Article  Google Scholar 

  4. Flusberg, B.A., Jung, J.C., Cocker, E.D., Anderson, E.P. & Schnitzer, M.J. Opt. Lett. 30, 2272–2274 (2005).

    Article  Google Scholar 

  5. Engelbrecht, C.J., Johnston, R.S., Seibel, E.J. & Helmchen, F. Opt. Express 16, 5556–5564 (2008).

    Article  CAS  Google Scholar 

  6. Flusberg, B.A. et al. Nat. Methods 5, 935–938 (2008).

    Article  CAS  Google Scholar 

  7. Hsiung, P.L. et al. Nat. Med. 14, 454–458 (2008).

    Article  CAS  Google Scholar 

  8. Llewellyn, M.E., Barretto, R.P., Delp, S.L. & Schnitzer, M.J. Nature 454, 784–788 (2008).

    Article  CAS  Google Scholar 

  9. Lin, K.Y., Maricevich, M., Bardeesy, N., Weissleder, R. & Mahmood, U. Transl. Oncol. 1, 84–94 (2008).

    Article  Google Scholar 

  10. Roulet, J.C. et al. Anal. Chem. 74, 3400–3407 (2002).

    Article  CAS  Google Scholar 

  11. Messerschmidt, B., McIntyre, B.L. & Houde-Walter, S.N. Appl. Opt. 35, 5670–5676 (1996).

    Article  CAS  Google Scholar 

  12. Sickenger, H., Falkenstoerfer, O.R., Lindlein, N. & Schwider, J. Opt. Eng. 33, 2680–2686 (1994).

    Article  Google Scholar 

  13. Born, M. & Wolf, E. Principles of Optics (Cambridge University Press, Cambridge, UK, 1999).

    Book  Google Scholar 

  14. Jung, J.C., Mehta, A.D., Aksay, E., Stepnoski, R. & Schnitzer, M.J. J. Neurophysiol. 92, 3121–3133 (2004).

    Article  Google Scholar 

  15. Knott, G. & Holtmaat, A. Brain Res. Rev. 58, 282–289 (2008).

    Article  Google Scholar 

  16. Jung, J.C. & Schnitzer, M.J. Opt. Lett. 28, 902–904 (2003).

    Article  Google Scholar 

  17. Messerschmidt, B., Possner, U. & Houde-Walter, S.N. Appl. Opt. 36, 8145–8152 (1997).

    Article  CAS  Google Scholar 

  18. Messerschmidt, B., Possner, T. & Goering, R. Appl. Opt. 34, 7825–7830 (1995).

    Article  CAS  Google Scholar 

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Acknowledgements

We thank the US National Institute of Neurological Disorders and Stroke (M.J.S.), GRINTech GmbH (B.M.) and the Stanford Biophysics (R.P.J.B.) training grant from the US National Institutes of Health for support of our work. We thank A. Attardo, L.D. Burns, E. Ho, J. Kobelke, S. Plochowietz and D. Profitt for technical assistance.

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Correspondence to Mark J Schnitzer.

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Barretto, R., Messerschmidt, B. & Schnitzer, M. In vivo fluorescence imaging with high-resolution microlenses. Nat Methods 6, 511–512 (2009). https://doi.org/10.1038/nmeth.1339

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