Abstract

Every optical imaging technique is limited in its penetration depth by scattering occurring in biological tissues. Possible solutions to overcome this problem consist of limiting the detrimental effects of scattering by reducing optical inhomogeneities within the sample. This can be achieved either by using physical methods (such as refractive index matching solutions) or by chemical methods (such as the removal of scatterers), based on tissue transformation protocols. This review provides an overview of the current state-of-the-art methods used for both ex-vivo and in-vivo optical clearing of biological tissues. We start with a brief history of the development of the most widespread clearing methods across the new millennium, then we describe the working principles of both physical and chemical methods. Clearing methods are then reviewed, pointing the attention of the reader on both physical and chemical methods, classified based on the tissue size and type for each specific application. A small section is reserved for methods that have already found in-vivo applications at the research level. Finally, a detailed discussion highlighting both the most relevant results achieved and the new ongoing developments in this field is reported in the last part, together with future perspectives for the clearing methodology.

© 2019 Optical Society of America under the terms of the OSA Open Access Publishing Agreement

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  90. R. Samatham, K. G. Phillips, and S. L. Jacques, “Assessment of Optical Clearing Agents Using Reflectance-Mode Confocal Scanning Laser Microscopy,” J. Innov. Opt. Health Sci. 3(03), 183–188 (2010).
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  91. Z. Mao, Z. Han, X. Wen, Q. Luo, and D. Zhu, “Influence of glycerol with different concentrations on skin optical clearing and morphological changes in vivo,” Proc. SPIE 7278, 72781I (2008).
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    [Crossref] [PubMed]
  93. R. Shi, L. Guo, C. Zhang, W. Feng, P. Li, Z. Ding, and D. Zhu, “A useful way to develop effective in vivo skin optical clearing agents,” J. Biophotonics 10(6-7), 887–895 (2017).
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  94. X. Q. Xu and Q. H. Zhu, “Sonophoretic delivery for contrast and depth improvement in skin optical coherence tomography,” IEEE J Sel Top Quant 14(1), 56–61 (2008).
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    [Crossref] [PubMed]
  96. S. A. Gallo, A. Sen, M. L. Hensen, and S. W. Hui, “Time-dependent ultrastructural changes to porcine stratum corneum following an electric pulse,” Biophys. J. 76(5), 2824–2832 (1999).
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  97. A. Sen, Y. Zhao, L. Zhang, and S. W. Hui, “Enhanced transdermal transport by electroporation using anionic lipids,” J. Control. Release 82(2-3), 399–405 (2002).
    [Crossref] [PubMed]
  98. S. A. Gallo, A. Sen, M. L. Hensen, and S. W. Hui, “Temperature-dependent electrical and ultrastructural characterizations of porcine skin upon electroporation,” Biophys. J. 82(1 Pt 1), 109–119 (2002).
    [Crossref] [PubMed]
  99. U. Pliquett, S. Gallo, S. W. Hui, Ch. Gusbeth, and E. Neumann, “Local and transient structural changes in stratum corneum at high electric fields: contribution of Joule heating,” Bioelectrochemistry 67(1), 37–46 (2005).
    [Crossref] [PubMed]
  100. X. Q. Xu and Q. H. Zhu, “Evaluation of skin optical clearing enhancement with Azone as a penetration enhancer,” Opt. Commun. 279(1), 223–228 (2007).
    [Crossref]
  101. R. A. McClure, C. Stoianovici, S. Karma, and B. Choi, “Revisiting Optical Clearing with Dimethyl Sulfoxide (DMSO): In Vitro and In Vivo Studies,” Proc SPIE 7187, 718707 (2009).
  102. K. D. Peck, A. H. Ghanem, and W. I. Higuchi, “Hindered Diffusion of Polar Molecules through and Effective Pore Radii Estimates of Intact and Ethanol Treated Human Epidermal Membrane,” Pharm. Res. 11(9), 1306–1314 (1994).
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  103. J. Wang, R. Shi, and D. Zhu, “Switchable skin window induced by optical clearing method for dermal blood flow imaging,” J. Biomed. Opt. 18(6), 061209 (2012).
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2019 (2)

Y. Qi, T. Yu, J. Xu, P. Wan, Y. Ma, J. Zhu, Y. Li, H. Gong, Q. Luo, and D. Zhu, “FDISCO: Advanced solvent-based clearing method for imaging whole organs,” Sci. Adv. 5(1), u8355 (2019).
[Crossref] [PubMed]

X. Y. Liu and B. Chen, “In Vivo Experimental Study on the Enhancement of Optical Clearing Effect by Laser Irradiation in Conjunction with a Chemical Penetration Enhancer,” Appl. Sci. (Basel) 9(3), 542 (2019).
[Crossref]

2018 (8)

Y. J. Zhao, T. T. Yu, C. Zhang, Z. Li, Q. M. Luo, T. H. Xu, and D. Zhu, “Skull optical clearing window for in vivo imaging of the mouse cortex at synaptic resolution,” Light Sci. Appl. 7(2), 17153 (2018).
[Crossref]

K. I. Sereti, N. B. Nguyen, P. Kamran, P. Zhao, S. Ranjbarvaziri, S. Park, S. Sabri, J. L. Engel, K. Sung, R. P. Kulkarni, Y. Ding, T. K. Hsiai, K. Plath, J. Ernst, D. Sahoo, H. K. A. Mikkola, M. L. Iruela-Arispe, and R. Ardehali, “Analysis of cardiomyocyte clonal expansion during mouse heart development and injury,” Nat. Commun. 9(1), 754 (2018).
[Crossref] [PubMed]

D. Jing, S. Zhang, W. Luo, X. Gao, Y. Men, C. Ma, X. Liu, Y. Yi, A. Bugde, B. O. Zhou, Z. Zhao, Q. Yuan, J. Q. Feng, L. Gao, W. P. Ge, and H. Zhao, “Tissue clearing of both hard and soft tissue organs with the PEGASOS method,” Cell Res. 28(8), 803–818 (2018).
[Crossref] [PubMed]

T. Yu, J. Zhu, Y. Li, Y. Ma, J. Wang, X. Cheng, S. Jin, Q. Sun, X. Li, H. Gong, Q. Luo, F. Xu, S. Zhao, and D. Zhu, “RTF: a rapid and versatile tissue optical clearing method,” Sci. Rep. 8(1), 1964 (2018).
[Crossref] [PubMed]

A. P. Di Giovanna, A. Tibo, L. Silvestri, M. C. Müllenbroich, I. Costantini, A. L. Allegra Mascaro, L. Sacconi, P. Frasconi, and F. S. Pavone, “Whole-Brain Vasculature Reconstruction at the Single Capillary Level,” Sci. Rep. 8(1), 12573 (2018).
[Crossref] [PubMed]

Y.-G. Park, C. H. Sohn, R. Chen, M. McCue, D. H. Yun, G. T. Drummond, T. Ku, N. B. Evans, H. C. Oak, W. Trieu, H. Choi, X. Jin, V. Lilascharoen, J. Wang, M. C. Truttmann, H. W. Qi, H. L. Ploegh, T. R. Golub, S.-C. Chen, M. P. Frosch, H. J. Kulik, B. K. Lim, and K. Chung, “Protection of tissue physicochemical properties using polyfunctional crosslinkers,” Nat. Biotechnol. 37, 73–83 (2018).
[PubMed]

A. Y. Sdobnov, M. E. Darvin, E. A. Genina, A. N. Bashkatov, J. Lademann, and V. V. Tuchin, “Recent progress in tissue optical clearing for spectroscopic application,” Spectrochim. Acta A Mol. Biomol. Spectrosc. 197, 216–229 (2018).
[Crossref] [PubMed]

T. Yu, Y. Qi, H. Gong, Q. Luo, and D. Zhu, “Optical clearing for multiscale biological tissues,” J. Biophotonics 11(2), e201700187 (2018).
[Crossref] [PubMed]

2017 (1)

R. Shi, L. Guo, C. Zhang, W. Feng, P. Li, Z. Ding, and D. Zhu, “A useful way to develop effective in vivo skin optical clearing agents,” J. Biophotonics 10(6-7), 887–895 (2017).
[Crossref] [PubMed]

2016 (5)

R. Torres, H. Velazquez, J. J. Chang, M. J. Levene, G. Moeckel, G. V. Desir, and R. Safirstein, “Three-Dimensional Morphology by Multiphoton Microscopy with Clearing in a Model of Cisplatin-Induced CKD,” J. Am. Soc. Nephrol. 27(4), 1102–1112 (2016).
[Crossref] [PubMed]

L. Silvestri, I. Costantini, L. Sacconi, and F. S. Pavone, “Clearing of fixed tissue: a review from a microscopist’s perspective,” J. Biomed. Opt. 21(8), 081205 (2016).
[Crossref] [PubMed]

E. Olson, M. J. Levene, and R. Torres, “Multiphoton microscopy with clearing for three dimensional histology of kidney biopsies,” Biomed. Opt. Express 7(8), 3089–3096 (2016).
[Crossref] [PubMed]

L. Ye, W. E. Allen, K. R. Thompson, Q. Tian, B. Hsueh, C. Ramakrishnan, A. C. Wang, J. H. Jennings, A. Adhikari, C. H. Halpern, I. B. Witten, A. L. Barth, L. Luo, J. A. McNab, and K. Deisseroth, “Wiring and Molecular Features of Prefrontal Ensembles Representing Distinct Experiences,” Cell 165(7), 1776–1788 (2016).
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C. Pan, R. Cai, F. P. Quacquarelli, A. Ghasemigharagoz, A. Lourbopoulos, P. Matryba, N. Plesnila, M. Dichgans, F. Hellal, and A. Ertürk, “Shrinkage-mediated imaging of entire organs and organisms using uDISCO,” Nat. Methods 13(10), 859–867 (2016).
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2015 (11)

I. Costantini, J. P. Ghobril, A. P. Di Giovanna, A. L. Allegra Mascaro, L. Silvestri, M. C. Müllenbroich, L. Onofri, V. Conti, F. Vanzi, L. Sacconi, R. Guerrini, H. Markram, G. Iannello, and F. S. Pavone, “A versatile clearing agent for multi-modal brain imaging,” Sci. Rep. 5(1), 9808 (2015).
[Crossref] [PubMed]

Y. Aoyagi, R. Kawakami, H. Osanai, T. Hibi, and T. Nemoto, “A Rapid Optical Clearing Protocol Using 2,2′-Thiodiethanol for Microscopic Observation of Fixed Mouse Brain,” PLoS One 10(1), e0116280 (2015).
[Crossref] [PubMed]

B. Hou, D. Zhang, S. Zhao, M. Wei, Z. Yang, S. Wang, J. Wang, X. Zhang, B. Liu, L. Fan, Y. Li, Z. Qiu, C. Zhang, and T. Jiang, “Scalable and Dil-compatible optical clearance of the mammalian brain,” Front. Neuroanat. 9, 19 (2015).
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H. Hama, H. Hioki, K. Namiki, T. Hoshida, H. Kurokawa, F. Ishidate, T. Kaneko, T. Akagi, T. Saito, T. Saido, and A. Miyawaki, “ScaleS: an optical clearing palette for biological imaging,” Nat. Neurosci. 18(10), 1518–1529 (2015).
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F. Chen, P. W. Tillberg, and E. S. Boyden, “Optical imaging. Expansion microscopy,” Science 347(6221), 543–548 (2015).
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S. Y. Kim, J. H. Cho, E. Murray, N. Bakh, H. Choi, K. Ohn, L. Ruelas, A. Hubbert, M. McCue, S. L. Vassallo, P. J. Keller, and K. Chung, “Stochastic electrotransport selectively enhances the transport of highly electromobile molecules,” Proc. Natl. Acad. Sci. U.S.A. 112(46), E6274–E6283 (2015).
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E. Murray, J. H. Cho, D. Goodwin, T. Ku, J. Swaney, S. Y. Kim, H. Choi, Y. G. Park, J. Y. Park, A. Hubbert, M. McCue, S. Vassallo, N. Bakh, M. P. Frosch, V. J. Wedeen, H. S. Seung, and K. Chung, “Simple, Scalable Proteomic Imaging for High-Dimensional Profiling of Intact Systems,” Cell 163(6), 1500–1514 (2015).
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A. L. Allegra Mascaro, I. Costantini, E. Margoni, G. Iannello, A. Bria, L. Sacconi, and F. S. Pavone, “Label-free near-infrared reflectance microscopy as a complimentary tool for two-photon fluorescence brain imaging,” Biomed. Opt. Express 6(11), 4483–4492 (2015).
[Crossref] [PubMed]

D. S. Richardson and J. W. Lichtman, “Clarifying Tissue Clearing,” Cell 162(2), 246–257 (2015).
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M. C. Müllenbroich, L. Silvestri, L. Onofri, I. Costantini, M. V. Hoff, L. Sacconi, G. Iannello, and F. S. Pavone, “Comprehensive optical and data management infrastructure for high-throughput light-sheet microscopy of whole mouse brains,” Neurophotonics 2(4), 041404 (2015).
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M. K. Schwarz, A. Scherbarth, R. Sprengel, J. Engelhardt, P. Theer, and G. Giese, “Fluorescent-protein stabilization and high-resolution imaging of cleared, intact mouse brains,” PLoS One 10(5), e0124650 (2015).
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2014 (6)

N. Renier, Z. Wu, D. J. Simon, J. Yang, P. Ariel, and M. Tessier-Lavigne, “iDISCO: A Simple, Rapid method to Immunolabel Large Tissue Samples for Volume Imaging,” Cell 159(4), 896–910 (2014).
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R. Torres, S. Vesuna, and M. J. Levene, “High-resolution, 2- and 3-dimensional imaging of uncut, unembedded tissue biopsy samples,” Arch. Pathol. Lab. Med. 138(3), 395–402 (2014).
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E. A. Susaki, K. Tainaka, D. Perrin, F. Kishino, T. Tawara, T. M. Watanabe, C. Yokoyama, H. Onoe, M. Eguchi, S. Yamaguchi, T. Abe, H. Kiyonari, Y. Shimizu, A. Miyawaki, H. Yokota, and H. R. Ueda, “Whole-brain imaging with single-cell resolution using chemical cocktails and computational analysis,” Cell 157(3), 726–739 (2014).
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Z. Deng, L. Jing, N. Wu, P. Lv, X. Jiang, Q. Ren, and C. Li, “Viscous optical clearing agent for in vivo optical imaging,” J. Biomed. Opt. 19(7), 76019 (2014).
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R. Tomer, L. Ye, B. Hsueh, and K. Deisseroth, “Advanced CLARITY for rapid and high-resolution imaging of intact tissues,” Nat. Protoc. 9(7), 1682–1697 (2014).
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B. Yang, J. B. Treweek, R. P. Kulkarni, B. E. Deverman, C. K. Chen, E. Lubeck, S. Shah, L. Cai, and V. Gradinaru, “Single-Cell Phenotyping within Transparent Intact Tissue through Whole-Body Clearing,” Cell 158(4), 945–958 (2014).
[Crossref] [PubMed]

2013 (5)

T. Kuwajima, A. A. Sitko, P. Bhansali, C. Jurgens, W. Guido, and C. Mason, “ClearT: a detergent- and solvent-free clearing method for neuronal and non-neuronal tissue,” Development 140(6), 1364–1368 (2013).
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M. T. Ke, S. Fujimoto, and T. Imai, “SeeDB: a simple and morphology-preserving optical clearing agent for neuronal circuit reconstruction,” Nat. Neurosci. 16(8), 1154–1161 (2013).
[Crossref] [PubMed]

D. Zhu, K. V. Larin, Q. Luo, and V. V. Tuchin, “Recent progress in tissue optical clearing,” Laser Photonics Rev. 7(5), 732–757 (2013).
[Crossref] [PubMed]

K. Chung, J. Wallace, S. Y. Kim, S. Kalyanasundaram, A. S. Andalman, T. J. Davidson, J. J. Mirzabekov, K. A. Zalocusky, J. Mattis, A. K. Denisin, S. Pak, H. Bernstein, C. Ramakrishnan, L. Grosenick, V. Gradinaru, and K. Deisseroth, “Structural and molecular interrogation of intact biological systems,” Nature 497(7449), 332–337 (2013).
[Crossref] [PubMed]

S. L. Jacques, “Optical properties of biological tissues: a review,” Phys. Med. Biol. 58(11), R37–R61 (2013).
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2012 (4)

A. Ertürk, K. Becker, N. Jährling, C. P. Mauch, C. D. Hojer, J. G. Egen, F. Hellal, F. Bradke, M. Sheng, and H. U. Dodt, “Three-dimensional imaging of solvent-cleared organs using 3DISCO,” Nat. Protoc. 7(11), 1983–1995 (2012).
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K. Becker, N. Jährling, S. Saghafi, R. Weiler, and H. U. Dodt, “Chemical clearing and dehydration of GFP expressing mouse brains,” PLoS One 7(3), e33916 (2012).
[Crossref] [PubMed]

P. J. Keller and H. U. Dodt, “Light sheet microscopy of living or cleared specimens,” Curr. Opin. Neurobiol. 22(1), 138–143 (2012).
[Crossref] [PubMed]

J. Wang, R. Shi, and D. Zhu, “Switchable skin window induced by optical clearing method for dermal blood flow imaging,” J. Biomed. Opt. 18(6), 061209 (2012).
[Crossref] [PubMed]

2011 (2)

J. Wang, X. Zhou, S. Duan, Z. W. Chen, and D. Zhu, “Improvement of in vivo rat skin optical clearing with chemical penetration enhancers,” Proc. SPIE 7883, 78830Y (2011).
[Crossref]

H. Hama, H. Kurokawa, H. Kawano, R. Ando, T. Shimogori, H. Noda, K. Fukami, A. Sakaue-Sawano, and A. Miyawaki, “Scale: a chemical approach for fluorescence imaging and reconstruction of transparent mouse brain,” Nat. Neurosci. 14(11), 1481–1488 (2011).
[Crossref] [PubMed]

2010 (4)

D. Zhu, J. Wang, Z. Zhi, X. Wen, and Q. Luo, “Imaging dermal blood flow through the intact rat skin with an optical clearing method,” J. Biomed. Opt. 15(2), 026008 (2010).
[Crossref] [PubMed]

X. Wen, Z. Mao, Z. Han, V. V. Tuchin, and D. Zhu, “In vivo skin optical clearing by glycerol solutions: mechanism,” J. Biophotonics 3(1-2), 44–52 (2010).
[Crossref] [PubMed]

J. M. Hirshburg, K. M. Ravikumar, W. Hwang, and A. T. Yeh, “Molecular basis for optical clearing of collagenous tissues,” J. Biomed. Opt. 15(5), 055002 (2010).
[Crossref] [PubMed]

R. Samatham, K. G. Phillips, and S. L. Jacques, “Assessment of Optical Clearing Agents Using Reflectance-Mode Confocal Scanning Laser Microscopy,” J. Innov. Opt. Health Sci. 3(03), 183–188 (2010).
[Crossref]

2009 (1)

P. S. Tsai, J. P. Kaufhold, P. Blinder, B. Friedman, P. J. Drew, H. J. Karten, P. D. Lyden, and D. Kleinfeld, “Correlations of neuronal and microvascular densities in murine cortex revealed by direct counting and colocalization of nuclei and vessels,” J. Neurosci. 29(46), 14553–14570 (2009).
[Crossref] [PubMed]

2008 (3)

Z. Mao, Z. Han, X. Wen, Q. Luo, and D. Zhu, “Influence of glycerol with different concentrations on skin optical clearing and morphological changes in vivo,” Proc. SPIE 7278, 72781I (2008).
[Crossref]

X. Q. Xu and Q. H. Zhu, “Sonophoretic delivery for contrast and depth improvement in skin optical coherence tomography,” IEEE J Sel Top Quant 14(1), 56–61 (2008).
[Crossref]

J. Yoon, T. Son, E. H. Choi, B. Choi, J. S. Nelson, and B. Jung, “Enhancement of optical skin clearing efficacy using a microneedle roller,” J. Biomed. Opt. 13(2), 021103 (2008).
[Crossref] [PubMed]

2007 (4)

X. Q. Xu and Q. H. Zhu, “Evaluation of skin optical clearing enhancement with Azone as a penetration enhancer,” Opt. Commun. 279(1), 223–228 (2007).
[Crossref]

J. Hirshburg, B. Choi, J. S. Nelson, and A. T. Yeh, “Correlation between collagen solubility and skin optical clearing using sugars,” Lasers Surg. Med. 39(2), 140–144 (2007).
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T. Staudt, M. C. Lang, R. Medda, J. Engelhardt, and S. W. Hell, “2,2′-thiodiethanol: a new water soluble mounting medium for high resolution optical microscopy,” Microsc. Res. Tech. 70(1), 1–9 (2007).
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H. U. Dodt, U. Leischner, A. Schierloh, N. Jährling, C. P. Mauch, K. Deininger, J. M. Deussing, M. Eder, W. Zieglgänsberger, and K. Becker, “Ultramicroscopy: three-dimensional visualization of neuronal networks in the whole mouse brain,” Nat. Methods 4(4), 331–336 (2007).
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2006 (5)

V. V. Tuchin, G. B. Altshuler, A. A. Gavrilova, A. B. Pravdin, D. Tabatadze, J. Childs, and I. V. Yaroslavsky, “Optical clearing of skin using flash lamp-induced enhancement of epidermal permeability,” Lasers Surg. Med. 38(9), 824–836 (2006).
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S. R. Millon, K. M. Roldan-Perez, K. M. Riching, G. M. Palmer, and N. Ramanujam, “Effect of optical clearing agents on the in vivo optical properties of squamous epithelial tissue,” Lasers Surg. Med. 38(10), 920–927 (2006).
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A. T. Yeh and J. Hirshburg, “Molecular interactions of exogenous chemical agents with collagen--implications for tissue optical clearing,” J. Biomed. Opt. 11(1), 014003 (2006).
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J. Hirshburg, B. Choi, J. S. Nelson, and A. T. Yeh, “Collagen solubility correlates with skin optical clearing,” J. Biomed. Opt. 11(4), 040501 (2006).
[Crossref] [PubMed]

O. Stumpp, B. Chen, and A. J. Welch, “Using sandpaper for noninvasive transepidermal optical skin clearing agent delivery,” J. Biomed. Opt. 11(4), 041118 (2006).
[Crossref] [PubMed]

2005 (4)

U. Pliquett, S. Gallo, S. W. Hui, Ch. Gusbeth, and E. Neumann, “Local and transient structural changes in stratum corneum at high electric fields: contribution of Joule heating,” Bioelectrochemistry 67(1), 37–46 (2005).
[Crossref] [PubMed]

V. V. Tuchin, “Optical clearing of tissues and blood using the immersion method,” J. Phys. D Appl. Phys. 38(15), 2497–2518 (2005).
[Crossref]

R. Cicchi, D. Sampson, D. Massi, and F. Pavone, “Contrast and depth enhancement in two-photon microscopy of human skin ex vivo by use of optical clearing agents,” Opt. Express 13(7), 2337–2344 (2005).
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B. Choi, L. Tsu, E. Chen, T. S. Ishak, S. M. Iskandar, S. Chess, and J. S. Nelson, “Determination of chemical agent optical clearing potential using in vitro human skin,” Lasers Surg. Med. 36(2), 72–75 (2005).
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2004 (3)

J. Huisken, J. Swoger, F. Del Bene, J. Wittbrodt, and E. H. Stelzer, “Optical sectioning deep inside live embryos by selective plane illumination microscopy,” Science 305(5686), 1007–1009 (2004).
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Y. He and R. K. K. Wang, “Dynamic optical clearing effect of tissue impregnated with hyperosmotic agents and studied with optical coherence tomography,” J. Biomed. Opt. 9(1), 200–206 (2004).
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S. N. Murthy, A. Sen, and S. W. Hui, “Surfactant-enhanced transdermal delivery by electroporation,” J. Control. Release 98(2), 307–315 (2004).
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2003 (4)

J. Gehl, “Electroporation: theory and methods, perspectives for drug delivery, gene therapy and research,” Acta Physiol. Scand. 177(4), 437–447 (2003).
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A. T. Yeh, B. Choi, J. S. Nelson, and B. J. Tromberg, “Reversible dissociation of collagen in tissues,” J. Invest. Dermatol. 121(6), 1332–1335 (2003).
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I. V. Meglinski, A. N. Bashkatov, E. A. Genina, D. Y. Churmakov, and V. V. Tuchin, “The enhancement of confocal images of tissues at bulk optical immersion,” Laser Phys. 13, 65–69 (2003).

X. Q. Xu, Y. H. He, S. Proskurin, R. K. K. Wang, and J. Elder, “Optical clearing of in vivo human skin with hyperosmotic chemicals investigated by optical coherence tomography and near infrared reflectance spectroscopy,” Proc. SPIE 5486, 129–135 (2003).

2002 (5)

A. F. Zuluaga, R. Drezek, T. Collier, R. Lotan, M. Follen, and R. Richards-Kortum, “Contrast agents for confocal microscopy: how simple chemicals affect confocal images of normal and cancer cells in suspension,” J. Biomed. Opt. 7(3), 398–403 (2002).
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A. V. Badkar and A. K. Banga, “Electrically enhanced transdermal delivery of a macromolecule,” J. Pharm. Pharmacol. 54(7), 907–912 (2002).
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P. G. Johnson, S. W. Hui, and A. R. Oseroff, “Electrically enhanced percutaneous delivery of delta-aminolevulinic acid using electric pulses and a DC potential,” Photochem. Photobiol. 75(5), 534–540 (2002).
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A. Sen, Y. Zhao, L. Zhang, and S. W. Hui, “Enhanced transdermal transport by electroporation using anionic lipids,” J. Control. Release 82(2-3), 399–405 (2002).
[Crossref] [PubMed]

S. A. Gallo, A. Sen, M. L. Hensen, and S. W. Hui, “Temperature-dependent electrical and ultrastructural characterizations of porcine skin upon electroporation,” Biophys. J. 82(1 Pt 1), 109–119 (2002).
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2001 (3)

R. K. K. Wang, X. Q. Xu, V. V. Tuchin, and J. B. Elder, “Concurrent enhancement of imaging depth and contrast for optical coherence tomography by hyperosmotic agents,” J. Opt. Soc. Am. B 18(7), 948–953 (2001).
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H. Steinke and W. Wolff, “A modified Spalteholz technique with preservation of the histology,” Ann. Anat. 183(1), 91–95 (2001).
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V. V. Tuchin, A. N. Bashkatov, E. A. Genina, Y. P. Sinichkin, and N. A. Lakodina, “In vivo investigation of the immersion-liquid-induced human skin clearing dynamics,” Tech. Phys. Lett. 27(6), 489–490 (2001).
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2000 (3)

A. K. Dunn, V. P. Wallace, M. Coleno, M. W. Berns, and B. J. Tromberg, “Influence of optical properties on two-photon fluorescence imaging in turbid samples,” Appl. Opt. 39(7), 1194–1201 (2000).
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M. Gu, X. S. Gan, A. Kisteman, and M. G. Xu, “Comparison of penetration depth between two-photon excitation and single-photon excitation in imaging through turbid tissue media,” Appl. Phys. Lett. 77(10), 1551–1553 (2000).
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C. Lombry, N. Dujardin, and V. Préat, “Transdermal delivery of macromolecules using skin electroporation,” Pharm. Res. 17(1), 32–37 (2000).
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1999 (3)

R. Vanbever and V. Préat, “In vivo efficacy and safety of skin electroporation,” Adv. Drug Deliv. Rev. 35(1), 77–88 (1999).
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S. A. Gallo, A. Sen, M. L. Hensen, and S. W. Hui, “Time-dependent ultrastructural changes to porcine stratum corneum following an electric pulse,” Biophys. J. 76(5), 2824–2832 (1999).
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G. Vargas, E. K. Chan, J. K. Barton, H. G. Rylander, and A. J. Welch, “Use of an agent to reduce scattering in skin,” Lasers Surg. Med. 24(2), 133–141 (1999).
[Crossref] [PubMed]

1998 (1)

Y. A. Chizmadzhev, A. V. Indenbom, P. I. Kuzmin, S. V. Galichenko, J. C. Weaver, and R. O. Potts, “Electrical properties of skin at moderate voltages: Contribution of appendageal macropores,” Biophys. J. 74(2 Pt 1), 843–856 (1998).
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1997 (1)

V. V. Tuchin, I. L. Maksimova, D. A. Zimnyakov, I. L. Kon, A. H. Mavlyutov, and A. A. Mishin, “Light propagation in tissues with controlled optical properties,” J. Biomed. Opt. 2(4), 401–417 (1997).
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1996 (4)

H. Liu, B. Beauvoit, M. Kimura, and B. Chance, “Dependence of tissue optical properties on solute-induced changes in refractive index and osmolarity,” J. Biomed. Opt. 1(2), 200–211 (1996).
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V. V. Tuchin, I. L. Maksimova, D. A. Zimnyakov, I. L. Kon, A. K. Mavlutov, and A. A. Mishin, “Light propagation in tissues with controlled optical properties,” P Soc Photo-Opt Ins 2925, 118–142 (1996).

X. J. Wang, T. E. Milner, M. C. Chang, and J. S. Nelson, “Group refractive index measurement of dry and hydrated type I collagen films using optical low-coherence reflectometry,” J. Biomed. Opt. 1(2), 212–216 (1996).
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M. R. Prausnitz, J. A. Gimm, R. H. Guy, R. Langer, J. C. Weaver, and C. Cullander, “Imaging regions of transport across human stratum corneum during high-voltage and low-voltage exposures,” J. Pharm. Sci. 85(12), 1363–1370 (1996).
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1995 (1)

B. Chance, H. Liu, T. Kitai, and Y. Zhang, “Effects of solutes on optical properties of biological materials: models, cells, and tissues,” Anal. Biochem. 227(2), 351–362 (1995).
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1994 (1)

K. D. Peck, A. H. Ghanem, and W. I. Higuchi, “Hindered Diffusion of Polar Molecules through and Effective Pore Radii Estimates of Intact and Ethanol Treated Human Epidermal Membrane,” Pharm. Res. 11(9), 1306–1314 (1994).
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1990 (1)

W. F. Cheong, S. A. Prahl, and A. J. Welch, “A Review of the Optical Properties of Biological Tissues,” IEEE J. Quantum Electron. 26(12), 2166–2185 (1990).
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1957 (1)

Abe, T.

E. A. Susaki, K. Tainaka, D. Perrin, F. Kishino, T. Tawara, T. M. Watanabe, C. Yokoyama, H. Onoe, M. Eguchi, S. Yamaguchi, T. Abe, H. Kiyonari, Y. Shimizu, A. Miyawaki, H. Yokota, and H. R. Ueda, “Whole-brain imaging with single-cell resolution using chemical cocktails and computational analysis,” Cell 157(3), 726–739 (2014).
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Adhikari, A.

L. Ye, W. E. Allen, K. R. Thompson, Q. Tian, B. Hsueh, C. Ramakrishnan, A. C. Wang, J. H. Jennings, A. Adhikari, C. H. Halpern, I. B. Witten, A. L. Barth, L. Luo, J. A. McNab, and K. Deisseroth, “Wiring and Molecular Features of Prefrontal Ensembles Representing Distinct Experiences,” Cell 165(7), 1776–1788 (2016).
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Akagi, T.

H. Hama, H. Hioki, K. Namiki, T. Hoshida, H. Kurokawa, F. Ishidate, T. Kaneko, T. Akagi, T. Saito, T. Saido, and A. Miyawaki, “ScaleS: an optical clearing palette for biological imaging,” Nat. Neurosci. 18(10), 1518–1529 (2015).
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Albanese, A.

T. Ku, J. Swaney, J. Y. Park, A. Albanese, E. Murray, J. H. Cho, Y. G. Park, V. Mangena, J. P. Chen, and K. H. Chung, “Multiplexed and scalable super-resolution imaging of three-dimensional protein localization in size-adjustable tissues,” Nat Biotechnol 34, 973 (2016).

Allegra Mascaro, A. L.

A. P. Di Giovanna, A. Tibo, L. Silvestri, M. C. Müllenbroich, I. Costantini, A. L. Allegra Mascaro, L. Sacconi, P. Frasconi, and F. S. Pavone, “Whole-Brain Vasculature Reconstruction at the Single Capillary Level,” Sci. Rep. 8(1), 12573 (2018).
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I. Costantini, J. P. Ghobril, A. P. Di Giovanna, A. L. Allegra Mascaro, L. Silvestri, M. C. Müllenbroich, L. Onofri, V. Conti, F. Vanzi, L. Sacconi, R. Guerrini, H. Markram, G. Iannello, and F. S. Pavone, “A versatile clearing agent for multi-modal brain imaging,” Sci. Rep. 5(1), 9808 (2015).
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A. L. Allegra Mascaro, I. Costantini, E. Margoni, G. Iannello, A. Bria, L. Sacconi, and F. S. Pavone, “Label-free near-infrared reflectance microscopy as a complimentary tool for two-photon fluorescence brain imaging,” Biomed. Opt. Express 6(11), 4483–4492 (2015).
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Allen, W. E.

L. Ye, W. E. Allen, K. R. Thompson, Q. Tian, B. Hsueh, C. Ramakrishnan, A. C. Wang, J. H. Jennings, A. Adhikari, C. H. Halpern, I. B. Witten, A. L. Barth, L. Luo, J. A. McNab, and K. Deisseroth, “Wiring and Molecular Features of Prefrontal Ensembles Representing Distinct Experiences,” Cell 165(7), 1776–1788 (2016).
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Altshuler, G. B.

V. V. Tuchin, G. B. Altshuler, A. A. Gavrilova, A. B. Pravdin, D. Tabatadze, J. Childs, and I. V. Yaroslavsky, “Optical clearing of skin using flash lamp-induced enhancement of epidermal permeability,” Lasers Surg. Med. 38(9), 824–836 (2006).
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Andalman, A. S.

K. Chung, J. Wallace, S. Y. Kim, S. Kalyanasundaram, A. S. Andalman, T. J. Davidson, J. J. Mirzabekov, K. A. Zalocusky, J. Mattis, A. K. Denisin, S. Pak, H. Bernstein, C. Ramakrishnan, L. Grosenick, V. Gradinaru, and K. Deisseroth, “Structural and molecular interrogation of intact biological systems,” Nature 497(7449), 332–337 (2013).
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Ando, R.

H. Hama, H. Kurokawa, H. Kawano, R. Ando, T. Shimogori, H. Noda, K. Fukami, A. Sakaue-Sawano, and A. Miyawaki, “Scale: a chemical approach for fluorescence imaging and reconstruction of transparent mouse brain,” Nat. Neurosci. 14(11), 1481–1488 (2011).
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Aoyagi, Y.

Y. Aoyagi, R. Kawakami, H. Osanai, T. Hibi, and T. Nemoto, “A Rapid Optical Clearing Protocol Using 2,2′-Thiodiethanol for Microscopic Observation of Fixed Mouse Brain,” PLoS One 10(1), e0116280 (2015).
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Ardehali, R.

K. I. Sereti, N. B. Nguyen, P. Kamran, P. Zhao, S. Ranjbarvaziri, S. Park, S. Sabri, J. L. Engel, K. Sung, R. P. Kulkarni, Y. Ding, T. K. Hsiai, K. Plath, J. Ernst, D. Sahoo, H. K. A. Mikkola, M. L. Iruela-Arispe, and R. Ardehali, “Analysis of cardiomyocyte clonal expansion during mouse heart development and injury,” Nat. Commun. 9(1), 754 (2018).
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Ariel, P.

N. Renier, Z. Wu, D. J. Simon, J. Yang, P. Ariel, and M. Tessier-Lavigne, “iDISCO: A Simple, Rapid method to Immunolabel Large Tissue Samples for Volume Imaging,” Cell 159(4), 896–910 (2014).
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Badkar, A. V.

A. V. Badkar and A. K. Banga, “Electrically enhanced transdermal delivery of a macromolecule,” J. Pharm. Pharmacol. 54(7), 907–912 (2002).
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Bakh, N.

S. Y. Kim, J. H. Cho, E. Murray, N. Bakh, H. Choi, K. Ohn, L. Ruelas, A. Hubbert, M. McCue, S. L. Vassallo, P. J. Keller, and K. Chung, “Stochastic electrotransport selectively enhances the transport of highly electromobile molecules,” Proc. Natl. Acad. Sci. U.S.A. 112(46), E6274–E6283 (2015).
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E. Murray, J. H. Cho, D. Goodwin, T. Ku, J. Swaney, S. Y. Kim, H. Choi, Y. G. Park, J. Y. Park, A. Hubbert, M. McCue, S. Vassallo, N. Bakh, M. P. Frosch, V. J. Wedeen, H. S. Seung, and K. Chung, “Simple, Scalable Proteomic Imaging for High-Dimensional Profiling of Intact Systems,” Cell 163(6), 1500–1514 (2015).
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Banga, A. K.

A. V. Badkar and A. K. Banga, “Electrically enhanced transdermal delivery of a macromolecule,” J. Pharm. Pharmacol. 54(7), 907–912 (2002).
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Barer, R.

Barth, A. L.

L. Ye, W. E. Allen, K. R. Thompson, Q. Tian, B. Hsueh, C. Ramakrishnan, A. C. Wang, J. H. Jennings, A. Adhikari, C. H. Halpern, I. B. Witten, A. L. Barth, L. Luo, J. A. McNab, and K. Deisseroth, “Wiring and Molecular Features of Prefrontal Ensembles Representing Distinct Experiences,” Cell 165(7), 1776–1788 (2016).
[Crossref] [PubMed]

Barton, J. K.

G. Vargas, E. K. Chan, J. K. Barton, H. G. Rylander, and A. J. Welch, “Use of an agent to reduce scattering in skin,” Lasers Surg. Med. 24(2), 133–141 (1999).
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Bashkatov, A. N.

A. Y. Sdobnov, M. E. Darvin, E. A. Genina, A. N. Bashkatov, J. Lademann, and V. V. Tuchin, “Recent progress in tissue optical clearing for spectroscopic application,” Spectrochim. Acta A Mol. Biomol. Spectrosc. 197, 216–229 (2018).
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I. V. Meglinski, A. N. Bashkatov, E. A. Genina, D. Y. Churmakov, and V. V. Tuchin, “The enhancement of confocal images of tissues at bulk optical immersion,” Laser Phys. 13, 65–69 (2003).

V. V. Tuchin, A. N. Bashkatov, E. A. Genina, Y. P. Sinichkin, and N. A. Lakodina, “In vivo investigation of the immersion-liquid-induced human skin clearing dynamics,” Tech. Phys. Lett. 27(6), 489–490 (2001).
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Beauvoit, B.

H. Liu, B. Beauvoit, M. Kimura, and B. Chance, “Dependence of tissue optical properties on solute-induced changes in refractive index and osmolarity,” J. Biomed. Opt. 1(2), 200–211 (1996).
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Becker, K.

A. Ertürk, K. Becker, N. Jährling, C. P. Mauch, C. D. Hojer, J. G. Egen, F. Hellal, F. Bradke, M. Sheng, and H. U. Dodt, “Three-dimensional imaging of solvent-cleared organs using 3DISCO,” Nat. Protoc. 7(11), 1983–1995 (2012).
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K. Becker, N. Jährling, S. Saghafi, R. Weiler, and H. U. Dodt, “Chemical clearing and dehydration of GFP expressing mouse brains,” PLoS One 7(3), e33916 (2012).
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H. U. Dodt, U. Leischner, A. Schierloh, N. Jährling, C. P. Mauch, K. Deininger, J. M. Deussing, M. Eder, W. Zieglgänsberger, and K. Becker, “Ultramicroscopy: three-dimensional visualization of neuronal networks in the whole mouse brain,” Nat. Methods 4(4), 331–336 (2007).
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Berns, M. W.

Bernstein, H.

K. Chung, J. Wallace, S. Y. Kim, S. Kalyanasundaram, A. S. Andalman, T. J. Davidson, J. J. Mirzabekov, K. A. Zalocusky, J. Mattis, A. K. Denisin, S. Pak, H. Bernstein, C. Ramakrishnan, L. Grosenick, V. Gradinaru, and K. Deisseroth, “Structural and molecular interrogation of intact biological systems,” Nature 497(7449), 332–337 (2013).
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Bhansali, P.

T. Kuwajima, A. A. Sitko, P. Bhansali, C. Jurgens, W. Guido, and C. Mason, “ClearT: a detergent- and solvent-free clearing method for neuronal and non-neuronal tissue,” Development 140(6), 1364–1368 (2013).
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Blinder, P.

P. S. Tsai, J. P. Kaufhold, P. Blinder, B. Friedman, P. J. Drew, H. J. Karten, P. D. Lyden, and D. Kleinfeld, “Correlations of neuronal and microvascular densities in murine cortex revealed by direct counting and colocalization of nuclei and vessels,” J. Neurosci. 29(46), 14553–14570 (2009).
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Boyden, E. S.

F. Chen, P. W. Tillberg, and E. S. Boyden, “Optical imaging. Expansion microscopy,” Science 347(6221), 543–548 (2015).
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Bradke, F.

A. Ertürk, K. Becker, N. Jährling, C. P. Mauch, C. D. Hojer, J. G. Egen, F. Hellal, F. Bradke, M. Sheng, and H. U. Dodt, “Three-dimensional imaging of solvent-cleared organs using 3DISCO,” Nat. Protoc. 7(11), 1983–1995 (2012).
[Crossref] [PubMed]

Bria, A.

Bugde, A.

D. Jing, S. Zhang, W. Luo, X. Gao, Y. Men, C. Ma, X. Liu, Y. Yi, A. Bugde, B. O. Zhou, Z. Zhao, Q. Yuan, J. Q. Feng, L. Gao, W. P. Ge, and H. Zhao, “Tissue clearing of both hard and soft tissue organs with the PEGASOS method,” Cell Res. 28(8), 803–818 (2018).
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Cai, L.

B. Yang, J. B. Treweek, R. P. Kulkarni, B. E. Deverman, C. K. Chen, E. Lubeck, S. Shah, L. Cai, and V. Gradinaru, “Single-Cell Phenotyping within Transparent Intact Tissue through Whole-Body Clearing,” Cell 158(4), 945–958 (2014).
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Cai, R.

C. Pan, R. Cai, F. P. Quacquarelli, A. Ghasemigharagoz, A. Lourbopoulos, P. Matryba, N. Plesnila, M. Dichgans, F. Hellal, and A. Ertürk, “Shrinkage-mediated imaging of entire organs and organisms using uDISCO,” Nat. Methods 13(10), 859–867 (2016).
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Chan, E. K.

G. Vargas, E. K. Chan, J. K. Barton, H. G. Rylander, and A. J. Welch, “Use of an agent to reduce scattering in skin,” Lasers Surg. Med. 24(2), 133–141 (1999).
[Crossref] [PubMed]

Chance, B.

H. Liu, B. Beauvoit, M. Kimura, and B. Chance, “Dependence of tissue optical properties on solute-induced changes in refractive index and osmolarity,” J. Biomed. Opt. 1(2), 200–211 (1996).
[Crossref] [PubMed]

B. Chance, H. Liu, T. Kitai, and Y. Zhang, “Effects of solutes on optical properties of biological materials: models, cells, and tissues,” Anal. Biochem. 227(2), 351–362 (1995).
[Crossref] [PubMed]

Chang, J. J.

R. Torres, H. Velazquez, J. J. Chang, M. J. Levene, G. Moeckel, G. V. Desir, and R. Safirstein, “Three-Dimensional Morphology by Multiphoton Microscopy with Clearing in a Model of Cisplatin-Induced CKD,” J. Am. Soc. Nephrol. 27(4), 1102–1112 (2016).
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Chang, M. C.

X. J. Wang, T. E. Milner, M. C. Chang, and J. S. Nelson, “Group refractive index measurement of dry and hydrated type I collagen films using optical low-coherence reflectometry,” J. Biomed. Opt. 1(2), 212–216 (1996).
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Chen, B.

X. Y. Liu and B. Chen, “In Vivo Experimental Study on the Enhancement of Optical Clearing Effect by Laser Irradiation in Conjunction with a Chemical Penetration Enhancer,” Appl. Sci. (Basel) 9(3), 542 (2019).
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O. Stumpp, B. Chen, and A. J. Welch, “Using sandpaper for noninvasive transepidermal optical skin clearing agent delivery,” J. Biomed. Opt. 11(4), 041118 (2006).
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Chen, C. K.

B. Yang, J. B. Treweek, R. P. Kulkarni, B. E. Deverman, C. K. Chen, E. Lubeck, S. Shah, L. Cai, and V. Gradinaru, “Single-Cell Phenotyping within Transparent Intact Tissue through Whole-Body Clearing,” Cell 158(4), 945–958 (2014).
[Crossref] [PubMed]

Chen, E.

B. Choi, L. Tsu, E. Chen, T. S. Ishak, S. M. Iskandar, S. Chess, and J. S. Nelson, “Determination of chemical agent optical clearing potential using in vitro human skin,” Lasers Surg. Med. 36(2), 72–75 (2005).
[Crossref] [PubMed]

Chen, F.

F. Chen, P. W. Tillberg, and E. S. Boyden, “Optical imaging. Expansion microscopy,” Science 347(6221), 543–548 (2015).
[Crossref] [PubMed]

Chen, J. P.

T. Ku, J. Swaney, J. Y. Park, A. Albanese, E. Murray, J. H. Cho, Y. G. Park, V. Mangena, J. P. Chen, and K. H. Chung, “Multiplexed and scalable super-resolution imaging of three-dimensional protein localization in size-adjustable tissues,” Nat Biotechnol 34, 973 (2016).

Chen, R.

Y.-G. Park, C. H. Sohn, R. Chen, M. McCue, D. H. Yun, G. T. Drummond, T. Ku, N. B. Evans, H. C. Oak, W. Trieu, H. Choi, X. Jin, V. Lilascharoen, J. Wang, M. C. Truttmann, H. W. Qi, H. L. Ploegh, T. R. Golub, S.-C. Chen, M. P. Frosch, H. J. Kulik, B. K. Lim, and K. Chung, “Protection of tissue physicochemical properties using polyfunctional crosslinkers,” Nat. Biotechnol. 37, 73–83 (2018).
[PubMed]

Chen, S.-C.

Y.-G. Park, C. H. Sohn, R. Chen, M. McCue, D. H. Yun, G. T. Drummond, T. Ku, N. B. Evans, H. C. Oak, W. Trieu, H. Choi, X. Jin, V. Lilascharoen, J. Wang, M. C. Truttmann, H. W. Qi, H. L. Ploegh, T. R. Golub, S.-C. Chen, M. P. Frosch, H. J. Kulik, B. K. Lim, and K. Chung, “Protection of tissue physicochemical properties using polyfunctional crosslinkers,” Nat. Biotechnol. 37, 73–83 (2018).
[PubMed]

Chen, Z. W.

J. Wang, X. Zhou, S. Duan, Z. W. Chen, and D. Zhu, “Improvement of in vivo rat skin optical clearing with chemical penetration enhancers,” Proc. SPIE 7883, 78830Y (2011).
[Crossref]

Cheng, X.

T. Yu, J. Zhu, Y. Li, Y. Ma, J. Wang, X. Cheng, S. Jin, Q. Sun, X. Li, H. Gong, Q. Luo, F. Xu, S. Zhao, and D. Zhu, “RTF: a rapid and versatile tissue optical clearing method,” Sci. Rep. 8(1), 1964 (2018).
[Crossref] [PubMed]

Cheong, W. F.

W. F. Cheong, S. A. Prahl, and A. J. Welch, “A Review of the Optical Properties of Biological Tissues,” IEEE J. Quantum Electron. 26(12), 2166–2185 (1990).
[Crossref]

Chess, S.

B. Choi, L. Tsu, E. Chen, T. S. Ishak, S. M. Iskandar, S. Chess, and J. S. Nelson, “Determination of chemical agent optical clearing potential using in vitro human skin,” Lasers Surg. Med. 36(2), 72–75 (2005).
[Crossref] [PubMed]

Childs, J.

V. V. Tuchin, G. B. Altshuler, A. A. Gavrilova, A. B. Pravdin, D. Tabatadze, J. Childs, and I. V. Yaroslavsky, “Optical clearing of skin using flash lamp-induced enhancement of epidermal permeability,” Lasers Surg. Med. 38(9), 824–836 (2006).
[Crossref] [PubMed]

Chizmadzhev, Y. A.

Y. A. Chizmadzhev, A. V. Indenbom, P. I. Kuzmin, S. V. Galichenko, J. C. Weaver, and R. O. Potts, “Electrical properties of skin at moderate voltages: Contribution of appendageal macropores,” Biophys. J. 74(2 Pt 1), 843–856 (1998).
[Crossref] [PubMed]

Cho, J. H.

E. Murray, J. H. Cho, D. Goodwin, T. Ku, J. Swaney, S. Y. Kim, H. Choi, Y. G. Park, J. Y. Park, A. Hubbert, M. McCue, S. Vassallo, N. Bakh, M. P. Frosch, V. J. Wedeen, H. S. Seung, and K. Chung, “Simple, Scalable Proteomic Imaging for High-Dimensional Profiling of Intact Systems,” Cell 163(6), 1500–1514 (2015).
[Crossref] [PubMed]

S. Y. Kim, J. H. Cho, E. Murray, N. Bakh, H. Choi, K. Ohn, L. Ruelas, A. Hubbert, M. McCue, S. L. Vassallo, P. J. Keller, and K. Chung, “Stochastic electrotransport selectively enhances the transport of highly electromobile molecules,” Proc. Natl. Acad. Sci. U.S.A. 112(46), E6274–E6283 (2015).
[Crossref] [PubMed]

T. Ku, J. Swaney, J. Y. Park, A. Albanese, E. Murray, J. H. Cho, Y. G. Park, V. Mangena, J. P. Chen, and K. H. Chung, “Multiplexed and scalable super-resolution imaging of three-dimensional protein localization in size-adjustable tissues,” Nat Biotechnol 34, 973 (2016).

Choi, B.

J. Yoon, T. Son, E. H. Choi, B. Choi, J. S. Nelson, and B. Jung, “Enhancement of optical skin clearing efficacy using a microneedle roller,” J. Biomed. Opt. 13(2), 021103 (2008).
[Crossref] [PubMed]

J. Hirshburg, B. Choi, J. S. Nelson, and A. T. Yeh, “Correlation between collagen solubility and skin optical clearing using sugars,” Lasers Surg. Med. 39(2), 140–144 (2007).
[Crossref] [PubMed]

J. Hirshburg, B. Choi, J. S. Nelson, and A. T. Yeh, “Collagen solubility correlates with skin optical clearing,” J. Biomed. Opt. 11(4), 040501 (2006).
[Crossref] [PubMed]

B. Choi, L. Tsu, E. Chen, T. S. Ishak, S. M. Iskandar, S. Chess, and J. S. Nelson, “Determination of chemical agent optical clearing potential using in vitro human skin,” Lasers Surg. Med. 36(2), 72–75 (2005).
[Crossref] [PubMed]

A. T. Yeh, B. Choi, J. S. Nelson, and B. J. Tromberg, “Reversible dissociation of collagen in tissues,” J. Invest. Dermatol. 121(6), 1332–1335 (2003).
[Crossref] [PubMed]

Choi, E. H.

J. Yoon, T. Son, E. H. Choi, B. Choi, J. S. Nelson, and B. Jung, “Enhancement of optical skin clearing efficacy using a microneedle roller,” J. Biomed. Opt. 13(2), 021103 (2008).
[Crossref] [PubMed]

Choi, H.

Y.-G. Park, C. H. Sohn, R. Chen, M. McCue, D. H. Yun, G. T. Drummond, T. Ku, N. B. Evans, H. C. Oak, W. Trieu, H. Choi, X. Jin, V. Lilascharoen, J. Wang, M. C. Truttmann, H. W. Qi, H. L. Ploegh, T. R. Golub, S.-C. Chen, M. P. Frosch, H. J. Kulik, B. K. Lim, and K. Chung, “Protection of tissue physicochemical properties using polyfunctional crosslinkers,” Nat. Biotechnol. 37, 73–83 (2018).
[PubMed]

S. Y. Kim, J. H. Cho, E. Murray, N. Bakh, H. Choi, K. Ohn, L. Ruelas, A. Hubbert, M. McCue, S. L. Vassallo, P. J. Keller, and K. Chung, “Stochastic electrotransport selectively enhances the transport of highly electromobile molecules,” Proc. Natl. Acad. Sci. U.S.A. 112(46), E6274–E6283 (2015).
[Crossref] [PubMed]

E. Murray, J. H. Cho, D. Goodwin, T. Ku, J. Swaney, S. Y. Kim, H. Choi, Y. G. Park, J. Y. Park, A. Hubbert, M. McCue, S. Vassallo, N. Bakh, M. P. Frosch, V. J. Wedeen, H. S. Seung, and K. Chung, “Simple, Scalable Proteomic Imaging for High-Dimensional Profiling of Intact Systems,” Cell 163(6), 1500–1514 (2015).
[Crossref] [PubMed]

Chung, K.

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Supplementary Material (1)

NameDescription
» Data File 1       In this review an objective analysis of the applicability of the various clearing methods published during the last decades was done analyzing the literature in an unbiased way. We excluded papers related to technological development in either clear

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Figures (6)

Fig. 1
Fig. 1 Graph showing the trend of published papers on tissue clearing during the last century. Data obtained from Scopus (www.scopus.com). The recent developments of aqueous solutions by Tuchin and associates, as well as the development of tissue transformation techniques, provoked a high widespread of the field, identified by the two abrupt variations in slope occurring in the mid 90’s and 2013, respectively.
Fig. 2
Fig. 2 (top) Usage of different clearing methods in papers focused on specific biological applications. The center and right pie charts show usage of clearing methods in combination with light-sheet fluorescence microscopy and confocal laser-scanning microscopy, respectively. (bottom) Timeline of the use of different clearing techniques for biological applications. The details of the papers used for this literature analysis can be found in Data File 1.
Fig. 3
Fig. 3 (a) Whole mouse brain before and after FDISCO clearing. (b) Thy1-GFP-M mouse brain images with insets at high resolution (c) 3D visualization of the vasculature in the mouse brain labeled by injection of CD31-A647 antibody. Images modified with permission from Qi et al [44].
Fig. 4
Fig. 4 (a) Whole mouse heart before and after CLARITY clearing. (b) Representative fluorescent microscope images of Rainbow heart at a different stage of development E14.5, P1, and P21 expressing Cre under the control of early cardiovascular progenitor transcription factors Mesp1 and Nkx2.5. Images modified with permission from Sereti et al. [67].
Fig. 5
Fig. 5 In-vivo two-photon cortical imaging of mouse brain through an optically cleared intact skull. Before imaging, the skull was topically treated with 10% collagenase (10% EDTA for P21-P30 aged mice) for 5-10 minutes and then 80% glycerol was dropped onto the skull. (a) Orthogonal (x–z) projections of dendrites through the intact skull, before and after skull optical clearing, demonstrating that the depth is obviously enhanced after clearing (the imaging parameter and data processing were the same). Scale bar = 10 μm. (b) The depth when imaging the dendrites of Thy1-YFP neurons, before and after skull optical clearing (P30, n = 10 mice; statistical method: one-way analysis of variance (ANOVA); Po0.001). Images modified with permission from Zhao et al. [75].
Fig. 6
Fig. 6 Transdermal imaging of blood circulation in mouse through optically cleared skin using laser speckle contrast imaging (LSCI). LSCI provides low or high contrast depending if the imaged scatterers are moving or stationary, respectively. Optical clearing was obtained using topical application of polyethylene glycol (PEG-400) alone or in combination with 10% Thiazone as chemical enhancer. Photographs (top row) and laser speckle temporal contrast maps (bottom row) of in vivo rat skin at the initial state, 4, 12, 24, 40 min after treatment of different OCAs, and 2 min after treatment of saline. (a) PEG-400, (b) mixed solution of PEG-400 and Thiazone. (c) Dynamic temporal contrast in five specific areas: 1, 2 in a (treatment with PEG-400), 3, 4, and 5 in b (treatment with a mixture of PEG-400 and Thiazone), before and after the application of optical clearing agents and saline. Here, 1, 3, and 4 are vessel areas, while 2 and 5 are no-vessel areas. Images modified with permission from Zhu et al. [70].

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