Abstract

We introduce a new method of hyperspectral imaging which encodes a varying temporal intensity modulation onto the excitation (or illumination) power spectrum. Functionally, we have moved the spectrometer from the back end of an experiment to the front, where intensity modulations uniquely label wavelengths within the excitation power spectrum at different frequencies, thereby creating a temporal light label which we can identify after subsequent light–matter interactions. To demonstrate this method, we acquire two-dimensional micrographs of background-free absorption spectra by capturing the intensity modulations transferred from the excitation spectrum into the emitted fluorescent intensity. Both the temporal light labeling method and the demonstrated excitation-labeled fluorescence application are readily adaptable to hyperspectral acquisition rates far beyond the frame rates of high-speed cameras.

© 2015 Optical Society of America

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References

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  1. D. G. Ferris, R. A. Lawhead, E. D. Dickman, N. Holtzapple, J. A. Miller, S. Grogan, S. Bambot, A. Agrawal, and M. L. Faupel, J. Low. Genit. Tract Dis. 5, 65 (2001).
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    [Crossref]
  4. T. Vo-Dinh, Biomedical Photonics Handbook: Biomedical Diagnostics (CRC Press, 2014), Vol. 2.
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    [Crossref]
  6. D. Abookasis, C. C. Lay, M. S. Mathews, M. E. Linskey, R. D. Frostig, and B. J. Tromberg, J. Biomed. Opt. 14, 024033 (2009).
    [Crossref]
  7. G. Lu and B. Fei, J. Biomed. Opt. 19, 10901 (2014).
    [Crossref]
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    [Crossref]
  9. D. G. Winters and R. A. Bartels, Opt. Lett. 40, 2774 (2015).
    [Crossref]
  10. D. J. Higley, D. G. Winters, G. L. Futia, and R. A. Bartels, J. Opt. Soc. Am. A 29, 2579 (2012).
    [Crossref]
  11. E. D. Diebold, B. W. Buckley, D. R. Gossett, and B. Jalali, Nat. Photonics 7, 806 (2013).
    [Crossref]
  12. C.-S. Liao, M. N. Slipchenko, P. Wang, J. Li, S.-Y. Lee, R. A. Oglesbee, and J.-X. Cheng, Light Sci. Appl. 4, e265 (2015).
    [Crossref]
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    [Crossref]
  14. S. R. Domingue and R. A. Bartels, Opt. Lett. 40, 253 (2015).
    [Crossref]
  15. E. Frumker, E. Tal, Y. Silberberg, and D. Majer, Opt. Lett. 30, 2796 (2005).
    [Crossref]

2015 (3)

C.-S. Liao, M. N. Slipchenko, P. Wang, J. Li, S.-Y. Lee, R. A. Oglesbee, and J.-X. Cheng, Light Sci. Appl. 4, e265 (2015).
[Crossref]

S. R. Domingue and R. A. Bartels, Opt. Lett. 40, 253 (2015).
[Crossref]

D. G. Winters and R. A. Bartels, Opt. Lett. 40, 2774 (2015).
[Crossref]

2014 (1)

G. Lu and B. Fei, J. Biomed. Opt. 19, 10901 (2014).
[Crossref]

2013 (2)

E. D. Diebold, B. W. Buckley, D. R. Gossett, and B. Jalali, Nat. Photonics 7, 806 (2013).
[Crossref]

Y. Zhang, Y. Chen, Y. Yu, X. Xue, V. V. Tuchin, and D. Zhu, J. Biomed. Opt. 18, 077003 (2013).
[Crossref]

2012 (1)

2011 (2)

G. Futia, P. Schlup, D. G. Winters, and R. A. Bartels, Opt. Express 19, 1626 (2011).
[Crossref]

J. R. Weber, D. J. Cuccia, W. R. Johnson, G. H. Bearman, A. J. Durkin, M. Hsu, A. Lin, D. K. Binder, D. Wilson, and B. J. Tromberg, J. Biomed. Opt. 16, 011015 (2011).
[Crossref]

2009 (1)

D. Abookasis, C. C. Lay, M. S. Mathews, M. E. Linskey, R. D. Frostig, and B. J. Tromberg, J. Biomed. Opt. 14, 024033 (2009).
[Crossref]

2005 (1)

2001 (1)

D. G. Ferris, R. A. Lawhead, E. D. Dickman, N. Holtzapple, J. A. Miller, S. Grogan, S. Bambot, A. Agrawal, and M. L. Faupel, J. Low. Genit. Tract Dis. 5, 65 (2001).

1999 (1)

1991 (1)

J. S. Sanders, R. G. Driggers, C. E. Halford, and S. T. Griffin, Opt. Eng. 30, 1720 (1991).
[Crossref]

Abookasis, D.

D. Abookasis, C. C. Lay, M. S. Mathews, M. E. Linskey, R. D. Frostig, and B. J. Tromberg, J. Biomed. Opt. 14, 024033 (2009).
[Crossref]

Agrawal, A.

D. G. Ferris, R. A. Lawhead, E. D. Dickman, N. Holtzapple, J. A. Miller, S. Grogan, S. Bambot, A. Agrawal, and M. L. Faupel, J. Low. Genit. Tract Dis. 5, 65 (2001).

Bambot, S.

D. G. Ferris, R. A. Lawhead, E. D. Dickman, N. Holtzapple, J. A. Miller, S. Grogan, S. Bambot, A. Agrawal, and M. L. Faupel, J. Low. Genit. Tract Dis. 5, 65 (2001).

Bartels, R. A.

Bearman, G. H.

J. R. Weber, D. J. Cuccia, W. R. Johnson, G. H. Bearman, A. J. Durkin, M. Hsu, A. Lin, D. K. Binder, D. Wilson, and B. J. Tromberg, J. Biomed. Opt. 16, 011015 (2011).
[Crossref]

Binder, D. K.

J. R. Weber, D. J. Cuccia, W. R. Johnson, G. H. Bearman, A. J. Durkin, M. Hsu, A. Lin, D. K. Binder, D. Wilson, and B. J. Tromberg, J. Biomed. Opt. 16, 011015 (2011).
[Crossref]

Buckley, B. W.

E. D. Diebold, B. W. Buckley, D. R. Gossett, and B. Jalali, Nat. Photonics 7, 806 (2013).
[Crossref]

Carder, K. L.

Chen, Y.

Y. Zhang, Y. Chen, Y. Yu, X. Xue, V. V. Tuchin, and D. Zhu, J. Biomed. Opt. 18, 077003 (2013).
[Crossref]

Cheng, J.-X.

C.-S. Liao, M. N. Slipchenko, P. Wang, J. Li, S.-Y. Lee, R. A. Oglesbee, and J.-X. Cheng, Light Sci. Appl. 4, e265 (2015).
[Crossref]

Cuccia, D. J.

J. R. Weber, D. J. Cuccia, W. R. Johnson, G. H. Bearman, A. J. Durkin, M. Hsu, A. Lin, D. K. Binder, D. Wilson, and B. J. Tromberg, J. Biomed. Opt. 16, 011015 (2011).
[Crossref]

Dickman, E. D.

D. G. Ferris, R. A. Lawhead, E. D. Dickman, N. Holtzapple, J. A. Miller, S. Grogan, S. Bambot, A. Agrawal, and M. L. Faupel, J. Low. Genit. Tract Dis. 5, 65 (2001).

Diebold, E. D.

E. D. Diebold, B. W. Buckley, D. R. Gossett, and B. Jalali, Nat. Photonics 7, 806 (2013).
[Crossref]

Domingue, S. R.

Driggers, R. G.

J. S. Sanders, R. G. Driggers, C. E. Halford, and S. T. Griffin, Opt. Eng. 30, 1720 (1991).
[Crossref]

Durkin, A. J.

J. R. Weber, D. J. Cuccia, W. R. Johnson, G. H. Bearman, A. J. Durkin, M. Hsu, A. Lin, D. K. Binder, D. Wilson, and B. J. Tromberg, J. Biomed. Opt. 16, 011015 (2011).
[Crossref]

Faupel, M. L.

D. G. Ferris, R. A. Lawhead, E. D. Dickman, N. Holtzapple, J. A. Miller, S. Grogan, S. Bambot, A. Agrawal, and M. L. Faupel, J. Low. Genit. Tract Dis. 5, 65 (2001).

Fei, B.

G. Lu and B. Fei, J. Biomed. Opt. 19, 10901 (2014).
[Crossref]

Ferris, D. G.

D. G. Ferris, R. A. Lawhead, E. D. Dickman, N. Holtzapple, J. A. Miller, S. Grogan, S. Bambot, A. Agrawal, and M. L. Faupel, J. Low. Genit. Tract Dis. 5, 65 (2001).

Frostig, R. D.

D. Abookasis, C. C. Lay, M. S. Mathews, M. E. Linskey, R. D. Frostig, and B. J. Tromberg, J. Biomed. Opt. 14, 024033 (2009).
[Crossref]

Frumker, E.

Futia, G.

Futia, G. L.

Gossett, D. R.

E. D. Diebold, B. W. Buckley, D. R. Gossett, and B. Jalali, Nat. Photonics 7, 806 (2013).
[Crossref]

Griffin, S. T.

J. S. Sanders, R. G. Driggers, C. E. Halford, and S. T. Griffin, Opt. Eng. 30, 1720 (1991).
[Crossref]

Grogan, S.

D. G. Ferris, R. A. Lawhead, E. D. Dickman, N. Holtzapple, J. A. Miller, S. Grogan, S. Bambot, A. Agrawal, and M. L. Faupel, J. Low. Genit. Tract Dis. 5, 65 (2001).

Halford, C. E.

J. S. Sanders, R. G. Driggers, C. E. Halford, and S. T. Griffin, Opt. Eng. 30, 1720 (1991).
[Crossref]

Higley, D. J.

Holtzapple, N.

D. G. Ferris, R. A. Lawhead, E. D. Dickman, N. Holtzapple, J. A. Miller, S. Grogan, S. Bambot, A. Agrawal, and M. L. Faupel, J. Low. Genit. Tract Dis. 5, 65 (2001).

Hsu, M.

J. R. Weber, D. J. Cuccia, W. R. Johnson, G. H. Bearman, A. J. Durkin, M. Hsu, A. Lin, D. K. Binder, D. Wilson, and B. J. Tromberg, J. Biomed. Opt. 16, 011015 (2011).
[Crossref]

Jalali, B.

E. D. Diebold, B. W. Buckley, D. R. Gossett, and B. Jalali, Nat. Photonics 7, 806 (2013).
[Crossref]

Johnson, W. R.

J. R. Weber, D. J. Cuccia, W. R. Johnson, G. H. Bearman, A. J. Durkin, M. Hsu, A. Lin, D. K. Binder, D. Wilson, and B. J. Tromberg, J. Biomed. Opt. 16, 011015 (2011).
[Crossref]

Lawhead, R. A.

D. G. Ferris, R. A. Lawhead, E. D. Dickman, N. Holtzapple, J. A. Miller, S. Grogan, S. Bambot, A. Agrawal, and M. L. Faupel, J. Low. Genit. Tract Dis. 5, 65 (2001).

Lay, C. C.

D. Abookasis, C. C. Lay, M. S. Mathews, M. E. Linskey, R. D. Frostig, and B. J. Tromberg, J. Biomed. Opt. 14, 024033 (2009).
[Crossref]

Lee, S.-Y.

C.-S. Liao, M. N. Slipchenko, P. Wang, J. Li, S.-Y. Lee, R. A. Oglesbee, and J.-X. Cheng, Light Sci. Appl. 4, e265 (2015).
[Crossref]

Lee, Z.

Li, J.

C.-S. Liao, M. N. Slipchenko, P. Wang, J. Li, S.-Y. Lee, R. A. Oglesbee, and J.-X. Cheng, Light Sci. Appl. 4, e265 (2015).
[Crossref]

Liao, C.-S.

C.-S. Liao, M. N. Slipchenko, P. Wang, J. Li, S.-Y. Lee, R. A. Oglesbee, and J.-X. Cheng, Light Sci. Appl. 4, e265 (2015).
[Crossref]

Lin, A.

J. R. Weber, D. J. Cuccia, W. R. Johnson, G. H. Bearman, A. J. Durkin, M. Hsu, A. Lin, D. K. Binder, D. Wilson, and B. J. Tromberg, J. Biomed. Opt. 16, 011015 (2011).
[Crossref]

Linskey, M. E.

D. Abookasis, C. C. Lay, M. S. Mathews, M. E. Linskey, R. D. Frostig, and B. J. Tromberg, J. Biomed. Opt. 14, 024033 (2009).
[Crossref]

Lu, G.

G. Lu and B. Fei, J. Biomed. Opt. 19, 10901 (2014).
[Crossref]

Majer, D.

Mathews, M. S.

D. Abookasis, C. C. Lay, M. S. Mathews, M. E. Linskey, R. D. Frostig, and B. J. Tromberg, J. Biomed. Opt. 14, 024033 (2009).
[Crossref]

Miller, J. A.

D. G. Ferris, R. A. Lawhead, E. D. Dickman, N. Holtzapple, J. A. Miller, S. Grogan, S. Bambot, A. Agrawal, and M. L. Faupel, J. Low. Genit. Tract Dis. 5, 65 (2001).

Mobley, C. D.

Oglesbee, R. A.

C.-S. Liao, M. N. Slipchenko, P. Wang, J. Li, S.-Y. Lee, R. A. Oglesbee, and J.-X. Cheng, Light Sci. Appl. 4, e265 (2015).
[Crossref]

Patch, J. S.

Sanders, J. S.

J. S. Sanders, R. G. Driggers, C. E. Halford, and S. T. Griffin, Opt. Eng. 30, 1720 (1991).
[Crossref]

Schlup, P.

Silberberg, Y.

Slipchenko, M. N.

C.-S. Liao, M. N. Slipchenko, P. Wang, J. Li, S.-Y. Lee, R. A. Oglesbee, and J.-X. Cheng, Light Sci. Appl. 4, e265 (2015).
[Crossref]

Steward, R. G.

Tal, E.

Tromberg, B. J.

J. R. Weber, D. J. Cuccia, W. R. Johnson, G. H. Bearman, A. J. Durkin, M. Hsu, A. Lin, D. K. Binder, D. Wilson, and B. J. Tromberg, J. Biomed. Opt. 16, 011015 (2011).
[Crossref]

D. Abookasis, C. C. Lay, M. S. Mathews, M. E. Linskey, R. D. Frostig, and B. J. Tromberg, J. Biomed. Opt. 14, 024033 (2009).
[Crossref]

Tuchin, V. V.

Y. Zhang, Y. Chen, Y. Yu, X. Xue, V. V. Tuchin, and D. Zhu, J. Biomed. Opt. 18, 077003 (2013).
[Crossref]

Vo-Dinh, T.

T. Vo-Dinh, Biomedical Photonics Handbook: Biomedical Diagnostics (CRC Press, 2014), Vol. 2.

Wang, P.

C.-S. Liao, M. N. Slipchenko, P. Wang, J. Li, S.-Y. Lee, R. A. Oglesbee, and J.-X. Cheng, Light Sci. Appl. 4, e265 (2015).
[Crossref]

Weber, J. R.

J. R. Weber, D. J. Cuccia, W. R. Johnson, G. H. Bearman, A. J. Durkin, M. Hsu, A. Lin, D. K. Binder, D. Wilson, and B. J. Tromberg, J. Biomed. Opt. 16, 011015 (2011).
[Crossref]

Wilson, D.

J. R. Weber, D. J. Cuccia, W. R. Johnson, G. H. Bearman, A. J. Durkin, M. Hsu, A. Lin, D. K. Binder, D. Wilson, and B. J. Tromberg, J. Biomed. Opt. 16, 011015 (2011).
[Crossref]

Winters, D. G.

Xue, X.

Y. Zhang, Y. Chen, Y. Yu, X. Xue, V. V. Tuchin, and D. Zhu, J. Biomed. Opt. 18, 077003 (2013).
[Crossref]

Yu, Y.

Y. Zhang, Y. Chen, Y. Yu, X. Xue, V. V. Tuchin, and D. Zhu, J. Biomed. Opt. 18, 077003 (2013).
[Crossref]

Zhang, Y.

Y. Zhang, Y. Chen, Y. Yu, X. Xue, V. V. Tuchin, and D. Zhu, J. Biomed. Opt. 18, 077003 (2013).
[Crossref]

Zhu, D.

Y. Zhang, Y. Chen, Y. Yu, X. Xue, V. V. Tuchin, and D. Zhu, J. Biomed. Opt. 18, 077003 (2013).
[Crossref]

Appl. Opt. (1)

J. Biomed. Opt. (4)

Y. Zhang, Y. Chen, Y. Yu, X. Xue, V. V. Tuchin, and D. Zhu, J. Biomed. Opt. 18, 077003 (2013).
[Crossref]

J. R. Weber, D. J. Cuccia, W. R. Johnson, G. H. Bearman, A. J. Durkin, M. Hsu, A. Lin, D. K. Binder, D. Wilson, and B. J. Tromberg, J. Biomed. Opt. 16, 011015 (2011).
[Crossref]

D. Abookasis, C. C. Lay, M. S. Mathews, M. E. Linskey, R. D. Frostig, and B. J. Tromberg, J. Biomed. Opt. 14, 024033 (2009).
[Crossref]

G. Lu and B. Fei, J. Biomed. Opt. 19, 10901 (2014).
[Crossref]

J. Low. Genit. Tract Dis. (1)

D. G. Ferris, R. A. Lawhead, E. D. Dickman, N. Holtzapple, J. A. Miller, S. Grogan, S. Bambot, A. Agrawal, and M. L. Faupel, J. Low. Genit. Tract Dis. 5, 65 (2001).

J. Opt. Soc. Am. A (1)

Light Sci. Appl. (1)

C.-S. Liao, M. N. Slipchenko, P. Wang, J. Li, S.-Y. Lee, R. A. Oglesbee, and J.-X. Cheng, Light Sci. Appl. 4, e265 (2015).
[Crossref]

Nat. Photonics (1)

E. D. Diebold, B. W. Buckley, D. R. Gossett, and B. Jalali, Nat. Photonics 7, 806 (2013).
[Crossref]

Opt. Eng. (1)

J. S. Sanders, R. G. Driggers, C. E. Halford, and S. T. Griffin, Opt. Eng. 30, 1720 (1991).
[Crossref]

Opt. Express (1)

Opt. Lett. (3)

Other (1)

T. Vo-Dinh, Biomedical Photonics Handbook: Biomedical Diagnostics (CRC Press, 2014), Vol. 2.

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Figures (3)

Fig. 1.
Fig. 1. Concept figure. The intensity modulation encoded onto the excitation power spectrum is transferred to the fluorescence via the absorbed spectrum. A Fourier transform of the temporal dynamics of the emission intensity, across all emission wavelengths, recovers the absorbed spectrum through the unique labeling of excitation wavelength to modulation frequency.
Fig. 2.
Fig. 2. (a) HI-ELF experimental schematic. (b) The mapping between spatial location in the focal plane of the Martinez stretcher to wavelength (black) and an analogous mapping with modulation frequency (blue), both are compared to theory (red circles), with the modulation frequency curve offset by 1 mm for visibility. (c) Excitation power spectrum entering the microscope as captured by a spectrometer with the reticle set to an angle with high modulation (blue) and by temporal LiLa (black).
Fig. 3.
Fig. 3. Image processing: (a) image hypercube in time domain, (b) single-pixel temporal data (blue) and gating filter (black), (c) fast Fourier transform of single-pixel temporal data (black) compared to the excitation spectrum (blue), and (d) single-pixel unabsorbed spectrum (black) compared to the reference unabsorbed spectra (Alexa Fluor 514 and 546, blue-dashed and blue-solid, respectively). Wide field micrographs of two lens tissues dyed with Alexa Fluor 514 and 546 using (e) fluorescence intensity only and (f) HI-ELF, Alexa Fluor 514 and 546 in blue and red, respectively.

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