Abstract

3-D two-photon excitation (TPE) microscopy has been a critical tool for biological study since its introduction. Yet, the speed is largely limited by its point detector, e.g., photomultiplier tube (PMT), which requires a point-scanning imaging sequence. In this Letter, we present a multi-focus compressive sensing (CS) method for 3-D and random-access TPE microscopy based on a digital micromirror device (DMD). This new platform combines CS with a unique holography-based DMD random-access scanner to enhance the imaging speed by three to five times for imaging arbitrarily selected regions in 3-D specimens without sacrificing the resolution. In the experiments, 1–20 randomly selected foci are generated by modulating the wavefront of a femtosecond laser via binary holography, where the combined intensity is recorded by a PMT. By exploiting CS algorithms, 3-D images at arbitrarily selected sites can be reconstructed. Simulations and imaging experiments on different samples have been performed to verify the principle and identify the optimal processing parameters, including the number of laser foci and sampling ratios. The results show that high-resolution images can be obtained by using a 25% sampling ratio and five foci. The new CS-based TPE imaging method may find important applications in biological studies, e.g., neuronal imaging and optogenetics.

© 2019 Optical Society of America

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References

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Bahlmann, K.

Bobin, J.

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Buehler, C.

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[Crossref]

Chahid, M.

V. Studer, J. Bobin, M. Chahid, H. S. Mousavi, E. Candes, and M. Dahan, Proc. Natl. Acad. Sci. USA 109, E1679 (2012).
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Chen, P.

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Chen, S.

Cheng, J.

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G. Katona, G. Szalay, P. Maák, A. Kaszás, M. Veress, D. Hillier, B. Chiovini, E. S. Vizi, B. Roska, and B. Rózsa, Nat. Methods 9, 201 (2012).
[Crossref]

Dahan, M.

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[Crossref]

Dong, C.-Y.

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M. P. Edgar, G. M. Gibson, and M. J. Padgett, Nat. Photonics 13, 13 (2019).
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Fantini, S.

Geng, Q.

Q. Geng, D. Wang, P. Chen, and S. Chen, Nat. Commun. 10, 2179 (2019).
[Crossref]

Q. Geng, C. Gu, J. Cheng, and S. Chen, Optica 4, 674 (2017).
[Crossref]

Gibson, G. M.

M. P. Edgar, G. M. Gibson, and M. J. Padgett, Nat. Photonics 13, 13 (2019).
[Crossref]

Gu, C.

Heffer, E. L.

Hillier, D.

G. Katona, G. Szalay, P. Maák, A. Kaszás, M. Veress, D. Hillier, B. Chiovini, E. S. Vizi, B. Roska, and B. Rózsa, Nat. Methods 9, 201 (2012).
[Crossref]

Hsu, L.

P. T. C. So, K. H. Kim, C. Buehler, B. R. Masters, L. Hsu, and C.-Y. Dong, Methods Cell. Imaging 14, 7 (2013).

Jiang, H.

C. Li, W. Yin, H. Jiang, and Y. Zhang, Comput. Optim. Appl. 56, 507 (2013).
[Crossref]

Jiang, J.

Kaszás, A.

G. Katona, G. Szalay, P. Maák, A. Kaszás, M. Veress, D. Hillier, B. Chiovini, E. S. Vizi, B. Roska, and B. Rózsa, Nat. Methods 9, 201 (2012).
[Crossref]

Katona, G.

G. Katona, G. Szalay, P. Maák, A. Kaszás, M. Veress, D. Hillier, B. Chiovini, E. S. Vizi, B. Roska, and B. Rózsa, Nat. Methods 9, 201 (2012).
[Crossref]

Ke, Y.

Kelemen, L.

Kim, K. H.

P. T. C. So, K. H. Kim, C. Buehler, B. R. Masters, L. Hsu, and C.-Y. Dong, Methods Cell. Imaging 14, 7 (2013).

K. H. Kim, C. Buehler, K. Bahlmann, T. Ragan, W.-C. A. Lee, E. Nedivi, E. L. Heffer, S. Fantini, and P. T. C. So, Opt. Express 15, 11658 (2007).
[Crossref]

Lee, W.-C. A.

Lee, W.-H.

Li, C.

C. Li, W. Yin, H. Jiang, and Y. Zhang, Comput. Optim. Appl. 56, 507 (2013).
[Crossref]

Maák, P.

G. Katona, G. Szalay, P. Maák, A. Kaszás, M. Veress, D. Hillier, B. Chiovini, E. S. Vizi, B. Roska, and B. Rózsa, Nat. Methods 9, 201 (2012).
[Crossref]

Mascaro, A. L. A.

Masters, B. R.

P. T. C. So, K. H. Kim, C. Buehler, B. R. Masters, L. Hsu, and C.-Y. Dong, Methods Cell. Imaging 14, 7 (2013).

Mousavi, H. S.

V. Studer, J. Bobin, M. Chahid, H. S. Mousavi, E. Candes, and M. Dahan, Proc. Natl. Acad. Sci. USA 109, E1679 (2012).
[Crossref]

Nedivi, E.

Ormos, P.

Padgett, M. J.

M. P. Edgar, G. M. Gibson, and M. J. Padgett, Nat. Photonics 13, 13 (2019).
[Crossref]

Pavone, F. S.

Ragan, T.

Roska, B.

G. Katona, G. Szalay, P. Maák, A. Kaszás, M. Veress, D. Hillier, B. Chiovini, E. S. Vizi, B. Roska, and B. Rózsa, Nat. Methods 9, 201 (2012).
[Crossref]

Rózsa, B.

G. Katona, G. Szalay, P. Maák, A. Kaszás, M. Veress, D. Hillier, B. Chiovini, E. S. Vizi, B. Roska, and B. Rózsa, Nat. Methods 9, 201 (2012).
[Crossref]

Sacconi, L.

Sancataldo, G.

Silvestri, L.

So, P. T. C.

P. T. C. So, K. H. Kim, C. Buehler, B. R. Masters, L. Hsu, and C.-Y. Dong, Methods Cell. Imaging 14, 7 (2013).

K. H. Kim, C. Buehler, K. Bahlmann, T. Ragan, W.-C. A. Lee, E. Nedivi, E. L. Heffer, S. Fantini, and P. T. C. So, Opt. Express 15, 11658 (2007).
[Crossref]

Studer, V.

V. Studer, J. Bobin, M. Chahid, H. S. Mousavi, E. Candes, and M. Dahan, Proc. Natl. Acad. Sci. USA 109, E1679 (2012).
[Crossref]

Szalay, G.

G. Katona, G. Szalay, P. Maák, A. Kaszás, M. Veress, D. Hillier, B. Chiovini, E. S. Vizi, B. Roska, and B. Rózsa, Nat. Methods 9, 201 (2012).
[Crossref]

Veress, M.

G. Katona, G. Szalay, P. Maák, A. Kaszás, M. Veress, D. Hillier, B. Chiovini, E. S. Vizi, B. Roska, and B. Rózsa, Nat. Methods 9, 201 (2012).
[Crossref]

Vizi, E. S.

G. Katona, G. Szalay, P. Maák, A. Kaszás, M. Veress, D. Hillier, B. Chiovini, E. S. Vizi, B. Roska, and B. Rózsa, Nat. Methods 9, 201 (2012).
[Crossref]

Vizsnyiczai, G.

Walker, S.

Wang, D.

Q. Geng, D. Wang, P. Chen, and S. Chen, Nat. Commun. 10, 2179 (2019).
[Crossref]

Yin, W.

C. Li, W. Yin, H. Jiang, and Y. Zhang, Comput. Optim. Appl. 56, 507 (2013).
[Crossref]

Yung, W. H.

Zhang, D.

Zhang, Y.

C. Li, W. Yin, H. Jiang, and Y. Zhang, Comput. Optim. Appl. 56, 507 (2013).
[Crossref]

Appl. Opt. (1)

Comput. Optim. Appl. (1)

C. Li, W. Yin, H. Jiang, and Y. Zhang, Comput. Optim. Appl. 56, 507 (2013).
[Crossref]

Methods Cell. Imaging (1)

P. T. C. So, K. H. Kim, C. Buehler, B. R. Masters, L. Hsu, and C.-Y. Dong, Methods Cell. Imaging 14, 7 (2013).

Nat. Commun. (1)

Q. Geng, D. Wang, P. Chen, and S. Chen, Nat. Commun. 10, 2179 (2019).
[Crossref]

Nat. Methods (1)

G. Katona, G. Szalay, P. Maák, A. Kaszás, M. Veress, D. Hillier, B. Chiovini, E. S. Vizi, B. Roska, and B. Rózsa, Nat. Methods 9, 201 (2012).
[Crossref]

Nat. Photonics (1)

M. P. Edgar, G. M. Gibson, and M. J. Padgett, Nat. Photonics 13, 13 (2019).
[Crossref]

Opt. Express (3)

Optica (2)

Proc. Natl. Acad. Sci. USA (1)

V. Studer, J. Bobin, M. Chahid, H. S. Mousavi, E. Candes, and M. Dahan, Proc. Natl. Acad. Sci. USA 109, E1679 (2012).
[Crossref]

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Figures (6)

Fig. 1.
Fig. 1. Optical configuration of the CS-based TPE microscope. M1, high-reflectivity mirror; L1–L4, lenses (fL1, fL2, fL3, fL4=100, 250, 200, and 200 mm, respectively); DM, dichroic mirror.
Fig. 2.
Fig. 2. (a) Simulated imaging experiments based on the Shepp–Logan phantom (top), mitochondria (middle), and pollen (bottom) with 1, 5, and 20 laser foci. (b) RE analysis of multi-focus CS imaging using 1–30 laser foci.
Fig. 3.
Fig. 3. Reflectance imaging results of parallel gold lines obtained in three different modes: (a) raster-scanning, (b) uniform under-sampling, and (c) 10-focus CS reconstruction. (d) Intensity profiles along the blue, green, and red lines in (a), (b), and (c), respectively. Scale bar = 5 μm.
Fig. 4.
Fig. 4. Reflectance imaging result of gold “CUHK” letters on a silicon substrate: (a) raster-scanning image; (b) CS images with a 30% sampling ratio and 2–20 foci; (c) CS images using 10 foci with 5%–30% sampling ratios. Scale bar = 10 μm.
Fig. 5.
Fig. 5. Cross-sectional TPE images of a pollen grain at five different depths obtained by (a) raster-scanning and (b) CS-based random-scanning with a 25% sampling ratio. Scale bar = 10 μm.
Fig. 6.
Fig. 6. Cross-sectional TPE images of a pollen grain on arbitrarily defined planes, i.e., parabolic, inclined, and sinusoidal surfaces: (a) raster-scanning and (b) CS imaging with a 25% sampling ratio.

Equations (8)

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

ϕ(x,y)=πΔzMobj2(x2+y2)λfL32+2πMobjλfL3(xΔx+yΔy),
H(i,j)={1,q2xT+ϕ(x,y)2π+kq20,otherwise,,
ϕ(x,y)=arg[twtei(ϕt(x,y)+θt)],
Vt=i=1mj=1nH(i,j)ei(ϕt(i·d,j·d)),
wt=wt|Vt||Vt|,
θt=arg(Vt),
b=Ax,
minxx1+τFx1s.t.b=Ax,