Abstract

Applying image subtraction strategy together with time-gated detection, we can reveal finer details in a fixed cell sample using moderate light intensities. The subtractive imaging is obtained from the subtraction of a suitably weighted short lifetime intensity-image from a long lifetime one, while the weight coefficient can be selected using the analysis of an optical transfer function. Compared with other subtractive methods, ours is regarded as an optimized variation of gated stimulated emission depletion microscopy and can be implemented in a simpler manner, providing lateral super-resolution, and it is theoretically possible to further expand its application scope.

© 2013 Optical Society of America

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References

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  1. J. B. Pawley, Handbook of Biological Confocal Microscopy (Plenum, 1990).
  2. M. Minsky, “Microscopy apparatus,” U. S. patent 3,013,467 (19December1961).
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    [CrossRef]
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    [CrossRef]
  5. M. Martinez-Corral, M. Caballero, E. H. K. Stelzer, and J. Swoger, Opt. Express 10, 98 (2002).
    [CrossRef]
  6. H. Okugawa, Proc. SPIE 6860, 68600K (2008).
    [CrossRef]
  7. C. B. Müller and J. Enderlein, Phys. Rev. Lett. 104, 198101 (2010).
    [CrossRef]
  8. E. Sánchez-Ortiga, C. J. R. Sheppard, G. Saavedra, M. Martínez-Corral, A. Doblas, and A. Calatayud, Opt. Lett. 37, 1280 (2012).
    [CrossRef]
  9. G. Vicidomini, G. Moneron, K. Y. Han, V. Westphal, H. Ta, M. Reuss, J. Engelhardt, C. Eggeling, and S. W. Hell, Nat. Methods 8, 571 (2011).
    [CrossRef]
  10. G. Vicidomini, A. Schönle, H. Ta, K. Y. Han, G. Moneron, C. Eggeling, and S. W. Hell, PLoS ONE 8, e54421 (2013).
    [CrossRef]
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    [CrossRef]
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    [CrossRef]
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    [CrossRef]
  14. R. Heintzmann, V. Sarafis, P. Munroe, J. Nailon, Q. S. Hanley, and T. M. Jovin, Micron 34, 293 (2003).
    [CrossRef]
  15. Z. Gu, C. Kuang, S. Li, Y. Xue, Z. Zheng, and X. Liu, Technical Digest (Optical Society of America, 2012), ATh2E.4.
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    [CrossRef]
  17. X. Hao, C. F. Kuang, T. T. Wang, and X. Liu, J. Opt. 12, 115707 (2010).
    [CrossRef]

2013 (1)

G. Vicidomini, A. Schönle, H. Ta, K. Y. Han, G. Moneron, C. Eggeling, and S. W. Hell, PLoS ONE 8, e54421 (2013).
[CrossRef]

2012 (3)

2011 (2)

T. Wilson, J. Microsc. 244, 113 (2011).
[CrossRef]

G. Vicidomini, G. Moneron, K. Y. Han, V. Westphal, H. Ta, M. Reuss, J. Engelhardt, C. Eggeling, and S. W. Hell, Nat. Methods 8, 571 (2011).
[CrossRef]

2010 (3)

X. Hao, C. F. Kuang, T. T. Wang, and X. Liu, J. Opt. 12, 115707 (2010).
[CrossRef]

L. Schermelleh, R. Heintzmann, and H. Leonhardt, J. Cell Biol. 190, 165 (2010).
[CrossRef]

C. B. Müller and J. Enderlein, Phys. Rev. Lett. 104, 198101 (2010).
[CrossRef]

2008 (1)

H. Okugawa, Proc. SPIE 6860, 68600K (2008).
[CrossRef]

2003 (2)

W. B. Amos and J. White, Mol. Biol. Cell 95, 335 (2003).
[CrossRef]

R. Heintzmann, V. Sarafis, P. Munroe, J. Nailon, Q. S. Hanley, and T. M. Jovin, Micron 34, 293 (2003).
[CrossRef]

2002 (1)

1959 (1)

B. Richards and E. Wolf, Proc. R. Soc. Lond. Ser. A Math. Phys. Eng. Sci. 253, 358 (1959).
[CrossRef]

Amos, W. B.

W. B. Amos and J. White, Mol. Biol. Cell 95, 335 (2003).
[CrossRef]

Benfenati, F.

Bianchini, P.

Caballero, M.

Calatayud, A.

Diaspro, A.

Doblas, A.

Eggeling, C.

G. Vicidomini, A. Schönle, H. Ta, K. Y. Han, G. Moneron, C. Eggeling, and S. W. Hell, PLoS ONE 8, e54421 (2013).
[CrossRef]

G. Vicidomini, G. Moneron, K. Y. Han, V. Westphal, H. Ta, M. Reuss, J. Engelhardt, C. Eggeling, and S. W. Hell, Nat. Methods 8, 571 (2011).
[CrossRef]

Enderlein, J.

C. B. Müller and J. Enderlein, Phys. Rev. Lett. 104, 198101 (2010).
[CrossRef]

Engelhardt, J.

G. Vicidomini, G. Moneron, K. Y. Han, V. Westphal, H. Ta, M. Reuss, J. Engelhardt, C. Eggeling, and S. W. Hell, Nat. Methods 8, 571 (2011).
[CrossRef]

Fišerová, E.

E. Fišerová and M. Kubala, J. Lumin. 132, 2059 (2012).
[CrossRef]

Galiani, S.

Gu, Z.

Z. Gu, C. Kuang, S. Li, Y. Xue, Z. Zheng, and X. Liu, Technical Digest (Optical Society of America, 2012), ATh2E.4.

Han, K. Y.

G. Vicidomini, A. Schönle, H. Ta, K. Y. Han, G. Moneron, C. Eggeling, and S. W. Hell, PLoS ONE 8, e54421 (2013).
[CrossRef]

G. Vicidomini, G. Moneron, K. Y. Han, V. Westphal, H. Ta, M. Reuss, J. Engelhardt, C. Eggeling, and S. W. Hell, Nat. Methods 8, 571 (2011).
[CrossRef]

Hanley, Q. S.

R. Heintzmann, V. Sarafis, P. Munroe, J. Nailon, Q. S. Hanley, and T. M. Jovin, Micron 34, 293 (2003).
[CrossRef]

Hao, X.

X. Hao, C. F. Kuang, T. T. Wang, and X. Liu, J. Opt. 12, 115707 (2010).
[CrossRef]

Harke, B.

Heintzmann, R.

L. Schermelleh, R. Heintzmann, and H. Leonhardt, J. Cell Biol. 190, 165 (2010).
[CrossRef]

R. Heintzmann, V. Sarafis, P. Munroe, J. Nailon, Q. S. Hanley, and T. M. Jovin, Micron 34, 293 (2003).
[CrossRef]

Hell, S. W.

G. Vicidomini, A. Schönle, H. Ta, K. Y. Han, G. Moneron, C. Eggeling, and S. W. Hell, PLoS ONE 8, e54421 (2013).
[CrossRef]

G. Vicidomini, G. Moneron, K. Y. Han, V. Westphal, H. Ta, M. Reuss, J. Engelhardt, C. Eggeling, and S. W. Hell, Nat. Methods 8, 571 (2011).
[CrossRef]

Jovin, T. M.

R. Heintzmann, V. Sarafis, P. Munroe, J. Nailon, Q. S. Hanley, and T. M. Jovin, Micron 34, 293 (2003).
[CrossRef]

Kuang, C.

Z. Gu, C. Kuang, S. Li, Y. Xue, Z. Zheng, and X. Liu, Technical Digest (Optical Society of America, 2012), ATh2E.4.

Kuang, C. F.

X. Hao, C. F. Kuang, T. T. Wang, and X. Liu, J. Opt. 12, 115707 (2010).
[CrossRef]

Kubala, M.

E. Fišerová and M. Kubala, J. Lumin. 132, 2059 (2012).
[CrossRef]

Leonhardt, H.

L. Schermelleh, R. Heintzmann, and H. Leonhardt, J. Cell Biol. 190, 165 (2010).
[CrossRef]

Li, S.

Z. Gu, C. Kuang, S. Li, Y. Xue, Z. Zheng, and X. Liu, Technical Digest (Optical Society of America, 2012), ATh2E.4.

Lignani, G.

Liu, X.

X. Hao, C. F. Kuang, T. T. Wang, and X. Liu, J. Opt. 12, 115707 (2010).
[CrossRef]

Z. Gu, C. Kuang, S. Li, Y. Xue, Z. Zheng, and X. Liu, Technical Digest (Optical Society of America, 2012), ATh2E.4.

Martinez-Corral, M.

Martínez-Corral, M.

Minsky, M.

M. Minsky, “Microscopy apparatus,” U. S. patent 3,013,467 (19December1961).

Moneron, G.

G. Vicidomini, A. Schönle, H. Ta, K. Y. Han, G. Moneron, C. Eggeling, and S. W. Hell, PLoS ONE 8, e54421 (2013).
[CrossRef]

G. Vicidomini, G. Moneron, K. Y. Han, V. Westphal, H. Ta, M. Reuss, J. Engelhardt, C. Eggeling, and S. W. Hell, Nat. Methods 8, 571 (2011).
[CrossRef]

Müller, C. B.

C. B. Müller and J. Enderlein, Phys. Rev. Lett. 104, 198101 (2010).
[CrossRef]

Munroe, P.

R. Heintzmann, V. Sarafis, P. Munroe, J. Nailon, Q. S. Hanley, and T. M. Jovin, Micron 34, 293 (2003).
[CrossRef]

Nailon, J.

R. Heintzmann, V. Sarafis, P. Munroe, J. Nailon, Q. S. Hanley, and T. M. Jovin, Micron 34, 293 (2003).
[CrossRef]

Okugawa, H.

H. Okugawa, Proc. SPIE 6860, 68600K (2008).
[CrossRef]

Pawley, J. B.

J. B. Pawley, Handbook of Biological Confocal Microscopy (Plenum, 1990).

Reuss, M.

G. Vicidomini, G. Moneron, K. Y. Han, V. Westphal, H. Ta, M. Reuss, J. Engelhardt, C. Eggeling, and S. W. Hell, Nat. Methods 8, 571 (2011).
[CrossRef]

Richards, B.

B. Richards and E. Wolf, Proc. R. Soc. Lond. Ser. A Math. Phys. Eng. Sci. 253, 358 (1959).
[CrossRef]

Saavedra, G.

Sánchez-Ortiga, E.

Sarafis, V.

R. Heintzmann, V. Sarafis, P. Munroe, J. Nailon, Q. S. Hanley, and T. M. Jovin, Micron 34, 293 (2003).
[CrossRef]

Schermelleh, L.

L. Schermelleh, R. Heintzmann, and H. Leonhardt, J. Cell Biol. 190, 165 (2010).
[CrossRef]

Schönle, A.

G. Vicidomini, A. Schönle, H. Ta, K. Y. Han, G. Moneron, C. Eggeling, and S. W. Hell, PLoS ONE 8, e54421 (2013).
[CrossRef]

Sheppard, C. J. R.

Stelzer, E. H. K.

Swoger, J.

Ta, H.

G. Vicidomini, A. Schönle, H. Ta, K. Y. Han, G. Moneron, C. Eggeling, and S. W. Hell, PLoS ONE 8, e54421 (2013).
[CrossRef]

G. Vicidomini, G. Moneron, K. Y. Han, V. Westphal, H. Ta, M. Reuss, J. Engelhardt, C. Eggeling, and S. W. Hell, Nat. Methods 8, 571 (2011).
[CrossRef]

Vicidomini, G.

G. Vicidomini, A. Schönle, H. Ta, K. Y. Han, G. Moneron, C. Eggeling, and S. W. Hell, PLoS ONE 8, e54421 (2013).
[CrossRef]

S. Galiani, B. Harke, G. Vicidomini, G. Lignani, F. Benfenati, A. Diaspro, and P. Bianchini, Opt. Express 20, 7362 (2012).
[CrossRef]

G. Vicidomini, G. Moneron, K. Y. Han, V. Westphal, H. Ta, M. Reuss, J. Engelhardt, C. Eggeling, and S. W. Hell, Nat. Methods 8, 571 (2011).
[CrossRef]

Wang, T. T.

X. Hao, C. F. Kuang, T. T. Wang, and X. Liu, J. Opt. 12, 115707 (2010).
[CrossRef]

Westphal, V.

G. Vicidomini, G. Moneron, K. Y. Han, V. Westphal, H. Ta, M. Reuss, J. Engelhardt, C. Eggeling, and S. W. Hell, Nat. Methods 8, 571 (2011).
[CrossRef]

White, J.

W. B. Amos and J. White, Mol. Biol. Cell 95, 335 (2003).
[CrossRef]

Wilson, T.

T. Wilson, J. Microsc. 244, 113 (2011).
[CrossRef]

Wolf, E.

B. Richards and E. Wolf, Proc. R. Soc. Lond. Ser. A Math. Phys. Eng. Sci. 253, 358 (1959).
[CrossRef]

Xue, Y.

Z. Gu, C. Kuang, S. Li, Y. Xue, Z. Zheng, and X. Liu, Technical Digest (Optical Society of America, 2012), ATh2E.4.

Zheng, Z.

Z. Gu, C. Kuang, S. Li, Y. Xue, Z. Zheng, and X. Liu, Technical Digest (Optical Society of America, 2012), ATh2E.4.

J. Cell Biol. (1)

L. Schermelleh, R. Heintzmann, and H. Leonhardt, J. Cell Biol. 190, 165 (2010).
[CrossRef]

J. Lumin. (1)

E. Fišerová and M. Kubala, J. Lumin. 132, 2059 (2012).
[CrossRef]

J. Microsc. (1)

T. Wilson, J. Microsc. 244, 113 (2011).
[CrossRef]

J. Opt. (1)

X. Hao, C. F. Kuang, T. T. Wang, and X. Liu, J. Opt. 12, 115707 (2010).
[CrossRef]

Micron (1)

R. Heintzmann, V. Sarafis, P. Munroe, J. Nailon, Q. S. Hanley, and T. M. Jovin, Micron 34, 293 (2003).
[CrossRef]

Mol. Biol. Cell (1)

W. B. Amos and J. White, Mol. Biol. Cell 95, 335 (2003).
[CrossRef]

Nat. Methods (1)

G. Vicidomini, G. Moneron, K. Y. Han, V. Westphal, H. Ta, M. Reuss, J. Engelhardt, C. Eggeling, and S. W. Hell, Nat. Methods 8, 571 (2011).
[CrossRef]

Opt. Express (2)

Opt. Lett. (1)

Phys. Rev. Lett. (1)

C. B. Müller and J. Enderlein, Phys. Rev. Lett. 104, 198101 (2010).
[CrossRef]

PLoS ONE (1)

G. Vicidomini, A. Schönle, H. Ta, K. Y. Han, G. Moneron, C. Eggeling, and S. W. Hell, PLoS ONE 8, e54421 (2013).
[CrossRef]

Proc. R. Soc. Lond. Ser. A Math. Phys. Eng. Sci. (1)

B. Richards and E. Wolf, Proc. R. Soc. Lond. Ser. A Math. Phys. Eng. Sci. 253, 358 (1959).
[CrossRef]

Proc. SPIE (1)

H. Okugawa, Proc. SPIE 6860, 68600K (2008).
[CrossRef]

Other (3)

J. B. Pawley, Handbook of Biological Confocal Microscopy (Plenum, 1990).

M. Minsky, “Microscopy apparatus,” U. S. patent 3,013,467 (19December1961).

Z. Gu, C. Kuang, S. Li, Y. Xue, Z. Zheng, and X. Liu, Technical Digest (Optical Society of America, 2012), ATh2E.4.

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Figures (4)

Fig. 1.
Fig. 1.

Graph of fluorescent lifetimes when ISTED= (a) 0, (b) 30, and (c) 250 mW, respectively.

Fig. 2.
Fig. 2.

Images of HEK cell taken with (a) a confocal microscope, (b) g-STED, and (c) our method, respectively. (d) Intensity profiles of the same region.

Fig. 3.
Fig. 3.

OTF of the (a) long lifetime, (b) short lifetime, and (c) subtractive intensity-images. The subtractive image is obtained when γ=0.5.

Fig. 4.
Fig. 4.

Intensity distributions of focal spots in axial plane when the incident beam is phase modulated (a) by a 02π helical phase plate or (b) by a 0/π circular one.

Equations (8)

Equations on this page are rendered with MathJax. Learn more.

heff=hconf·η(ISTED),
η(ISTED)exp(ISTEDTSTEDσSTED),
τfl=1/(kfl+σSTEDISTED),
hshort=0tgheffet/τfltdthlong=tgΔTheffet/τfltdt,
hsub=hlongγhshort,
OTFsub=OTFlongγOTFshort.
SNR(NlongγNshort)/Nlong+γNshort,
0<γ<(2Nlong+18Nlong+1)/2Nshort.

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