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High-magnification super-resolution FINCH microscopy using birefringent crystal lens interferometers

Nature Photonics volume 10, pages 802–808 (2016)Cite this article

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Abstract

Fresnel incoherent correlation holography (FINCH) microscopy is a promising approach for high-resolution biological imaging but has so far been limited to use with low-magnification, low-numerical-aperture configurations. We report the use of in-line incoherent interferometers made from uniaxial birefringent α-barium borate (α-BBO) or calcite crystals that overcome the aberrations and distortions present with previous implementations that employed spatial light modulators or gradient refractive index lenses. FINCH microscopy incorporating these birefringent elements and high-numerical-aperture oil immersion objectives could outperform standard wide-field fluorescence microscopy, with, for example, a 149 nm lateral point spread function at a wavelength of 590 nm. Enhanced resolution was confirmed with sub-resolution fluorescent beads. Taking the Golgi apparatus as a biological example, three different proteins labelled with GFP and two other fluorescent dyes in HeLa cells were resolved with an image quality that is comparable to similar samples captured by structured illumination microscopy.

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Figure 1: Conceptual schematic of a FINCH microscope.
Figure 2: Optical arrangement of birefringent lens incoherent interferometers.
Figure 3: Holograms from a birefringent lens-based calcite and GRIN lens FINCH system after expanded laser beam illumination.
Figure 4: Resolution comparison of wide-field and α-BBO FINCH imaging of fluorescent beads.
Figure 5: Comparative imaging of three different Golgi apparatus proteins in HeLa cells by wide-field and α-BBO FINCH.

References

  1. Gabor, D. A new microscopic principle. Nature 161, 777–778 (1948).

    Article  ADS  Google Scholar 

  2. Poon, T.-C. et al. Three-dimensional fluorescence microscopy by optical scanning holography. Opt. Eng. 34, 1338–1344 (1995).

    Article  ADS  Google Scholar 

  3. Rosen, J. & Brooker, G. Digital spatially incoherent Fresnel holography. Opt. Lett. 32, 912–914 (2007).

    Article  ADS  Google Scholar 

  4. Rosen, J. & Brooker, G. Non-scanning motionless fluorescence three-dimensional holographic microscopy. Nat. Photon. 2, 190–195 (2008).

    Article  ADS  Google Scholar 

  5. Brooker, G., Siegel, N., Wang, V. & Rosen, J. Optimal resolution in Fresnel incoherent correlation holographic fluorescence microscopy. Opt. Express 19, 5047–5062 (2011).

    Article  ADS  Google Scholar 

  6. Rosen, J., Siegel, N. & Brooker, G. Theoretical and experimental demonstration of resolution beyond the Rayleigh limit by FINCH fluorescence microscopic imaging. Opt. Express 19, 26249–26268 (2011).

    Article  ADS  Google Scholar 

  7. Katz, B., Rosen, J., Kelner, R. & Brooker, G. Enhanced resolution and throughput of Fresnel incoherent correlation holography (FINCH) using dual diffractive lenses on a spatial light modulator (SLM). Opt. Express 20, 9109–9121 (2012).

    Article  ADS  Google Scholar 

  8. Brooker, G. et al. In-line FINCH super resolution digital holographic fluorescence microscopy using a high efficiency transmission liquid crystal GRIN lens. Opt. Lett. 38, 5264–5267 (2013).

    Article  ADS  Google Scholar 

  9. Siegel, N. & Brooker, G. Improved axial resolution of FINCH fluorescence microscopy when combined with spinning disk confocal microscopy. Opt. Express 22, 22298–22307 (2014).

    Article  ADS  Google Scholar 

  10. Siegel, N., Rosen, J. & Brooker, G. Reconstruction of objects above and below the objective focal plane with dimensional fidelity by FINCH fluorescence microscopy. Opt. Express 20, 19822–19835 (2012).

    Article  ADS  Google Scholar 

  11. Siegel, N., Rosen, J. & Brooker, G. Faithful reconstruction of digital holograms captured by FINCH using a Hamming window function in the Fresnel propagation. Opt. Lett. 38, 3922–3925 (2013).

    Article  ADS  Google Scholar 

  12. Rosen, J. & Brooker, G. Fluorescence incoherent color holography. Opt. Express 15, 2244–2250 (2007).

    Article  ADS  Google Scholar 

  13. Kim, M. K. Adaptive optics by incoherent digital holography. Opt. Lett. 37, 2694 (2012).

    Article  ADS  Google Scholar 

  14. Yamaguchi, I. & Zhang, T. Phase-shifting digital holography. Opt. Lett. 22, 1268–1270 (1997).

    Article  ADS  Google Scholar 

  15. Dyson, J. Common-path interferometer for testing purposes. J. Opt. Soc. Am. A 47, 386–390 (1957).

    Article  ADS  Google Scholar 

  16. Goto, K., Sasaki, M., Okuma, S. & Hane, K. A double-focus lens interferometer for scanning force microscopy. Rev. Sci. Instrum. 66, 3182–3185 (1995).

    Article  ADS  Google Scholar 

  17. York, A. G. et al. Instant super-resolution imaging in live cells and embryos via analog image processing. Nat. Methods 10, 1122–1126 (2013).

    Article  Google Scholar 

  18. Cotte, Y. et al. Marker-free phase nanoscopy. Nat. Photon. 7, 113–117 (2013).

    Article  ADS  Google Scholar 

  19. Weng, J., Clark, D. C. & Kim, M. K. Compressive sensing sectional imaging for single-shot in-line self-interference incoherent holography. Opt. Commun. 366, 88–93 (2016).

    Article  ADS  Google Scholar 

  20. Pokrovskaya, I. D. et al. Conserved oligomeric Golgi complex specifically regulates the maintenance of Golgi glycosylation machinery. Glycobiology 21, 1554–1569 (2011).

    Article  Google Scholar 

Download references

Acknowledgements

This work was supported by CellOptic, Inc. and the National Institutes of Health under National Cancer Institute Award Number R44CA192299 and National Institute of General Medical Sciences Award Number U54GM105814. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. Patents have been applied for. We thank M. Bruce and M. Butte for assistance with the deconvolution.

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Authors and Affiliations

  1. Department of Biomedical Engineering, Johns Hopkins University, 9605 Medical Center Drive Suite 240, Rockville, 20850, Maryland, USA

    Nisan Siegel & Gary Brooker

  2. Microscopy Center, Johns Hopkins University Montgomery County Campus, Rockville, 20850, Maryland, USA

    Nisan Siegel & Gary Brooker

  3. CellOptic, Inc., 9605 Medical Center Drive Suite 224, Rockville, 20850, Maryland, USA

    Nisan Siegel & Gary Brooker

  4. Department of Physiology and Biophysics, University of Arkansas for Medical Science, 4301 West Markham Street, Little Rock, 72205, Arkansas, USA

    Vladimir Lupashin & Brian Storrie

Authors
  1. Nisan Siegel
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  2. Vladimir Lupashin
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  3. Brian Storrie
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  4. Gary Brooker
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Contributions

G.B. and N.S. designed and developed the concept for the birefringent lens interferometer and its implementation into the FINCH microscope, produced the hardware and software, conducted the FINCH and wide-field experiments and wrote the manuscript. V.L. prepared the HeLa cell immunolabelled fluorescent samples and took the image using the commercial SIM microscope shown in Supplementary Fig. 2. B.S. and V.L. provided various fluorescently labelled Golgi proteins in live cells and fixed samples during the course of this research that were important in the development of this new microscope. G.B. organized and supervised this work.

Corresponding author

Correspondence to Gary Brooker.

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Competing interests

The FINCH technology used in the study is owned by CellOptic, Inc. Additionally, this work was funded in part by CellOptic, Inc. G.B. is the founder of CellOptic, Inc., owns equity in the company and serves as the company's president and CEO. This arrangement has been reviewed and approved by the Johns Hopkins University in accordance with its conflict of interest policies. N.S. receives support from CellOptic, Inc. and B.S. is a consultant for CellOptic, Inc.

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Siegel, N., Lupashin, V., Storrie, B. et al. High-magnification super-resolution FINCH microscopy using birefringent crystal lens interferometers. Nature Photon 10, 802–808 (2016). https://doi.org/10.1038/nphoton.2016.207

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