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.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$259.00 per year
only $21.58 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Flusberg, B.A. et al. Nat. Methods 2, 941–950 (2005).
Levene, M.J., Dombeck, D.A., Kasischke, K.A., Molloy, R.P. & Webb, W.W. J. Neurophysiol. 91, 1908–1912 (2004).
Alencar, H., Mahmood, U., Kawano, Y., Hirata, T. & Weissleder, R. Neoplasia 7, 977–983 (2005).
Flusberg, B.A., Jung, J.C., Cocker, E.D., Anderson, E.P. & Schnitzer, M.J. Opt. Lett. 30, 2272–2274 (2005).
Engelbrecht, C.J., Johnston, R.S., Seibel, E.J. & Helmchen, F. Opt. Express 16, 5556–5564 (2008).
Flusberg, B.A. et al. Nat. Methods 5, 935–938 (2008).
Hsiung, P.L. et al. Nat. Med. 14, 454–458 (2008).
Llewellyn, M.E., Barretto, R.P., Delp, S.L. & Schnitzer, M.J. Nature 454, 784–788 (2008).
Lin, K.Y., Maricevich, M., Bardeesy, N., Weissleder, R. & Mahmood, U. Transl. Oncol. 1, 84–94 (2008).
Roulet, J.C. et al. Anal. Chem. 74, 3400–3407 (2002).
Messerschmidt, B., McIntyre, B.L. & Houde-Walter, S.N. Appl. Opt. 35, 5670–5676 (1996).
Sickenger, H., Falkenstoerfer, O.R., Lindlein, N. & Schwider, J. Opt. Eng. 33, 2680–2686 (1994).
Born, M. & Wolf, E. Principles of Optics (Cambridge University Press, Cambridge, UK, 1999).
Jung, J.C., Mehta, A.D., Aksay, E., Stepnoski, R. & Schnitzer, M.J. J. Neurophysiol. 92, 3121–3133 (2004).
Knott, G. & Holtmaat, A. Brain Res. Rev. 58, 282–289 (2008).
Jung, J.C. & Schnitzer, M.J. Opt. Lett. 28, 902–904 (2003).
Messerschmidt, B., Possner, U. & Houde-Walter, S.N. Appl. Opt. 36, 8145–8152 (1997).
Messerschmidt, B., Possner, T. & Goering, R. Appl. Opt. 34, 7825–7830 (1995).
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.
Author information
Authors and Affiliations
Corresponding author
Supplementary information
Supplementary Text and Figures
Supplementary Figures 1–2 (PDF 289 kb)
Rights and permissions
About this article
Cite this article
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
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/nmeth.1339
This article is cited by
-
Imagining the future of optical microscopy: everything, everywhere, all at once
Communications Biology (2023)
-
Two-Photon Endoscopy: State of the Art and Perspectives
Molecular Imaging and Biology (2023)
-
Bi-coloured enhanced luminescence imaging by targeted switch on/off laser MEF coupling for synthetic biosensing of nanostructured human serum albumin
Photochemical & Photobiological Sciences (2023)
-
High-speed light-sheet microscopy for the in-situ acquisition of volumetric histological images of living tissue
Nature Biomedical Engineering (2022)
-
Deep tissue multi-photon imaging using adaptive optics with direct focus sensing and shaping
Nature Biotechnology (2022)