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Nucleic acid and protein mass mapping by live-cell deep-ultraviolet microscopy

Nature Methods volume 4, pages 567–569 (2007)Cite this article

Abstract

We developed a deep-ultraviolet (UV) microscope capable of imaging cell mitosis and motility at 280 nm for 45 min with minimal UV-induced toxicity, and for 6 h before the onset of visible cell death in cultured human and mouse cells. Combined with computational methods that convert the intensity of each pixel into an estimate of mass, deep-UV microscopy images generate maps of nucleic acid mass, protein mass and fluorescence yield in unlabeled cells.

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Figure 1: Comparison of UV and visible-wavelength imaging.
Figure 2: Quantitative imaging of a fixed, unlabeled IC-21 cell.
Figure 3: Live-cell 280 nm UV imaging.

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Acknowledgements

We thank S. Inoué, M. Lang, D. Lauffenburger, S. Manalis and P. So for advice and encouragement, and J. Evans, Y. Freyzon, J. Hou, R. Lam, B. Sculimbrene and N. Watson for assistance with implementation. This work was supported by grants from the National Institutes of Health (P.M. and D.J.E.), and the Air Force Office of Scientific Research (D.J.E.), as well as support (to B.J.Z.) through the MIT Department of Biological Engineering including the Bernard E. Proctor Memorial Fellowship and the Viterbi Family Foundation Fund Fellowship.

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Authors and Affiliations

  1. WI-MIT BioImaging Center, 500 Technology Square NE47-287, Cambridge, 02142, Massachusetts, USA

    Benjamin J Zeskind, Caroline D Jordan, Winston Timp, Linda Trapani, Victor Horodincu & Paul Matsudaira

  2. Whitehead Institute for Biomedical Research, 9 Cambridge Center, Cambridge, 02142, Massachusetts, USA

    Benjamin J Zeskind, Winston Timp, Guichy Waller, Daniel J Ehrlich & Paul Matsudaira

  3. Department of Biological Engineering, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, 02139, Massachusetts, USA

    Benjamin J Zeskind & Paul Matsudaira

  4. Department of Mathematics, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, 02139, Massachusetts, USA

    Caroline D Jordan

  5. Departments of Electrical Engineering & Computer Science, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, 02139, Massachusetts, USA

    Winston Timp

  6. Department of Biology, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, 02139, Massachusetts, USA

    Paul Matsudaira

  7. Division of Graduate Medical Sciences, Boston University, 715 Albany Street L-317, Boston, 02118, Massachusetts, USA

    Linda Trapani

Authors
  1. Benjamin J Zeskind
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  7. Daniel J Ehrlich
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Corresponding author

Correspondence to Benjamin J Zeskind.

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Competing interests

Our institutions have applied for patents covering the methods and apparatus described in the manuscript.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–2, Supplementary Methods, Supplementary Discussion (PDF 1036 kb)

Supplementary Movie 1

Mitosis of a live HT-1080 (human epithelial fibrosarcoma) cell imaged in 280 nm transmission using 100 ms exposures separated by 1 minute of dark time. (MOV 1630 kb)

Supplementary Movie 2

Motility of a live IC-21(mouse macrophage) cell imaged in 280 nm transmission using 100 ms exposures separated by 1 minute of dark time. (MOV 2110 kb)

Supplementary Movie 3

Live HT-1080 (mouse macrophage) imaged in 280 nm transmission using 100 ms exposures separated by 1 minute of dark time, to illustrate dynamics of filopodia at high temporal resolution. (MOV 2197 kb)

Supplementary Movie 4

Live IC-21 (mouse macrophage) cell imaged for over 6 hours with 100 ms exposures separated by 1 minute of dark time. (MOV 2415 kb)

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Zeskind, B., Jordan, C., Timp, W. et al. Nucleic acid and protein mass mapping by live-cell deep-ultraviolet microscopy. Nat Methods 4, 567–569 (2007). https://doi.org/10.1038/nmeth1053

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