Abstract

Laser tattoo removal is an effective method of eliminating tattoo particles in the skin. However, laser treatment cannot always remove the unwanted tattoo completely, and there are risks of either temporary or permanent side effects. Studies using preclinical animal models could provide detailed information on the effects of laser treatment in the skin, and might help to minimize side effects in clinical practices. In this study, two-photon microscopy (TPM) was used to visualize the laser treatment effects on tattoo particles in both phantom specimens and in vivo mouse models. Fluorescent tattoo ink was used for particle visualization by TPM, and nanosecond (ns) and picosecond (ps) lasers at 532 nm were used for treatment. In phantom specimens, TPM characterized the fragmentation of individual tattoo particles by tracking them before and after the laser treatment. These changes were confirmed by field emission scanning electron microscopy (FE-SEM). TPM was used to measure the treatment efficiency of the two lasers at different laser fluences. In the mouse model, TPM visualized clusters of tattoo particles in the skin and detected their fragmentation after the laser treatment. Longitudinal TPM imaging observed the migration of cells containing tattoo particles after the laser treatment. These results show that TPM may be useful for the assessment of laser tattoo removal treatment in preclinical studies.

© 2017 Optical Society of America

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Corrections

8 August 2018: A typographical correction was made to the author affiliations.


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References

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    [Crossref] [PubMed]
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    [Crossref] [PubMed]
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    [Crossref] [PubMed]

2016 (5)

T. Wang, W. H. Jang, S. Lee, C. J. Yoon, J. H. Lee, B. Kim, S. Hwang, C. P. Hong, Y. Yoon, G. Lee, V. H. Le, S. Bok, G. O. Ahn, J. Lee, Y. S. Gho, E. Chung, S. Kim, M. H. Jang, S. J. Myung, M. J. Kim, P. T. So, and K. H. Kim, “Moxifloxacin: Clinically compatible contrast agent for multiphoton imaging,” Sci. Rep. 6(1), 27142 (2016).
[Crossref] [PubMed]

W. H. Jang, S. Shim, T. Wang, Y. Yoon, W. S. Jang, J. K. Myung, S. Park, and K. H. Kim, “In vivo characterization of early-stage radiation skin injury in a mouse model by two-photon microscopy,” Sci. Rep. 6(1), 19216 (2016).
[Crossref] [PubMed]

S. Lee, J. H. Lee, J. H. Park, Y. Yoon, W. K. Chung, H. Tchah, M. J. Kim, and K. H. Kim, “In vivo 3D measurement of moxifloxacin and gatifloxacin distributions in the mouse cornea using multiphoton microscopy,” Sci. Rep. 6(1), 25339 (2016).
[Crossref] [PubMed]

O. Reiter, L. Atzmony, L. Akerman, A. Levi, R. Kershenovich, M. Lapidoth, and D. Mimouni, “Picosecond lasers for tattoo removal: a systematic review,” Lasers Med. Sci. 31(7), 1397–1405 (2016).
[Crossref] [PubMed]

I. Saytashev, R. Glenn, G. A. Murashova, S. Osseiran, D. Spence, C. L. Evans, and M. Dantus, “Multiphoton excited hemoglobin fluorescence and third harmonic generation for non-invasive microscopy of stored blood,” Biomed. Opt. Express 7(9), 3449–3460 (2016).
[Crossref] [PubMed]

2015 (6)

S. J. Aurangabadkar, “Shifting paradigm in laser tattoo removal,” J. Cutan. Aesthet. Surg. 8(1), 3–4 (2015).
[Crossref] [PubMed]

S. D. Shah and S. J. Aurangabadkar, “Newer trends in laser tattoo removal,” J. Cutan. Aesthet. Surg. 8(1), 25–29 (2015).
[Crossref] [PubMed]

K. Sardana, R. Ranjan, and S. Ghunawat, “Optimising laser tattoo removal,” J. Cutan. Aesthet. Surg. 8(1), 16–24 (2015).
[Crossref] [PubMed]

N. Khunger, A. Molpariya, and A. Khunger, “Complications of tattoos and tattoo removal: Stop and think before you ink,” J. Cutan. Aesthet. Surg. 8(1), 30–36 (2015).
[Crossref] [PubMed]

F.-J. Leu, C.-L. Huang, Y.-M. Sue, S.-C. Lee, and C.-C. Wang, “Effects of tattoo ink’s absorption spectra and particle size on cosmetic tattoo treatment efficacy using Q-switched Nd:YAG laser,” Lasers Med. Sci. 30(1), 303–309 (2015).
[Crossref] [PubMed]

M. Balu, C. B. Zachary, R. M. Harris, T. B. Krasieva, K. König, B. J. Tromberg, and K. M. Kelly, “In vivo multiphoton microscopy of basal cell carcinoma,” JAMA Dermatol. 151(10), 1068–1074 (2015).
[Crossref] [PubMed]

2014 (1)

A. Klein, I. Rittmann, K.-A. Hiller, M. Landthaler, and W. Bäumler, “An Internet-based survey on characteristics of laser tattoo removal and associated side effects,” Lasers Med. Sci. 29(2), 729–738 (2014).
[Crossref] [PubMed]

2013 (2)

M. R. Tsai, C. Y. Lin, Y. H. Liao, and C. K. Sun, “Applying tattoo dye as a third-harmonic generation contrast agent for in vivo optical virtual biopsy of human skin,” J. Biomed. Opt. 18(2), 026012 (2013).
[Crossref] [PubMed]

B. G. Yipp and P. Kubes, “Antibodies against neutrophil LY6G do not inhibit leukocyte recruitment in mice in vivo,” Blood 121(1), 241–242 (2013).
[Crossref] [PubMed]

2012 (1)

N. Saedi, A. Metelitsa, K. Petrell, K. A. Arndt, and J. S. Dover, “Treatment of tattoos with a picosecond alexandrite laser: a prospective trial,” Arch. Dermatol. 148(12), 1360–1363 (2012).
[Crossref] [PubMed]

2011 (1)

L. G. Ng, J. S. Qin, B. Roediger, Y. Wang, R. Jain, L. L. Cavanagh, A. L. Smith, C. A. Jones, M. de Veer, M. A. Grimbaldeston, E. N. Meeusen, and W. Weninger, “Visualizing the neutrophil response to sterile tissue injury in mouse dermis reveals a three-phase cascade of events,” J. Invest. Dermatol. 131(10), 2058–2068 (2011).
[Crossref] [PubMed]

2010 (2)

S. Choudhary, M. L. Elsaie, A. Leiva, and K. Nouri, “Lasers for tattoo removal: a review,” Lasers Med. Sci. 25(5), 619–627 (2010).
[Crossref] [PubMed]

L. Izikson, W. Farinelli, F. Sakamoto, Z. Tannous, and R. R. Anderson, “Safety and effectiveness of black tattoo clearance in a pig model after a single treatment with a novel 758 nm 500 picosecond laser: a pilot study,” Lasers Surg. Med. 42(7), 640–646 (2010).
[Crossref] [PubMed]

2009 (2)

W. Kirby, A. Desai, T. Desai, F. Kartono, and P. Geeta, “The Kirby-Desai Scale: A Proposed Scale to Assess Tattoo-removal Treatments,” J. Clin. Aesthet. Dermatol. 2(3), 32–37 (2009).
[PubMed]

F. R. Ayers, D. J. Cuccia, K. M. Kelly, and A. J. Durkin, “Wide-field spatial mapping of in vivo tattoo skin optical properties using modulated imaging,” Lasers Surg. Med. 41(6), 442–453 (2009).
[Crossref] [PubMed]

2007 (1)

H. Morsy, M. Mogensen, L. Thrane, and G. B. Jemec, “Imaging of intradermal tattoos by optical coherence tomography,” Skin Res. Technol. 13(4), 444–448 (2007).
[Crossref] [PubMed]

2006 (3)

K. O’goshi, C. Suihko, and J. Serup, “In vivo imaging of intradermal tattoos by confocal scanning laser microscopy,” Skin Res. Technol. 12(2), 94–98 (2006).
[Crossref] [PubMed]

D. Débarre, W. Supatto, A.-M. Pena, A. Fabre, T. Tordjmann, L. Combettes, M.-C. Schanne-Klein, and E. Beaurepaire, “Imaging lipid bodies in cells and tissues using third-harmonic generation microscopy,” Nat. Methods 3(1), 47–53 (2006).
[Crossref] [PubMed]

J. M. Handley, “Adverse events associated with nonablative cutaneous visible and infrared laser treatment,” J. Am. Acad. Dermatol. 55(3), 482–489 (2006).
[Crossref] [PubMed]

2005 (1)

2004 (1)

T. R. Mempel, S. E. Henrickson, and U. H. Von Andrian, “T-cell priming by dendritic cells in lymph nodes occurs in three distinct phases,” Nature 427(6970), 154–159 (2004).
[Crossref] [PubMed]

2003 (1)

W. R. Zipfel, R. M. Williams, and W. W. Webb, “Nonlinear magic: multiphoton microscopy in the biosciences,” Nat. Biotechnol. 21(11), 1369–1377 (2003).
[Crossref] [PubMed]

2002 (1)

D. D. Ho, R. London, G. B. Zimmerman, and D. A. Young, “Laser-tattoo removal--a study of the mechanism and the optimal treatment strategy via computer simulations,” Lasers Surg. Med. 30(5), 389–397 (2002).
[Crossref] [PubMed]

1999 (1)

M. Rajadhyaksha, S. González, J. M. Zavislan, R. R. Anderson, and R. H. Webb, “In vivo confocal scanning laser microscopy of human skin II: advances in instrumentation and comparison with histology,” J. Invest. Dermatol. 113(3), 293–303 (1999).
[Crossref] [PubMed]

1998 (3)

J. Squier, M. Müller, G. Brakenhoff, and K. R. Wilson, “Third harmonic generation microscopy,” Opt. Express 3(9), 315–324 (1998).
[Crossref] [PubMed]

V. Ross, G. Naseef, G. Lin, M. Kelly, N. Michaud, T. J. Flotte, J. Raythen, and R. R. Anderson, “Comparison of responses of tattoos to picosecond and nanosecond Q-switched neodymium: YAG lasers,” Arch. Dermatol. 134(2), 167–171 (1998).
[Crossref] [PubMed]

V. Ross, G. Naseef, G. Lin, M. Kelly, N. Michaud, T. J. Flotte, J. Raythen, and R. R. Anderson, “Comparison of responses of tattoos to picosecond and nanosecond Q-switched neodymium: YAG lasers,” Arch. Dermatol. 134(2), 167–171 (1998).
[Crossref] [PubMed]

1997 (2)

B. R. Masters, P. T. So, and E. Gratton, “Multiphoton excitation fluorescence microscopy and spectroscopy of in vivo human skin,” Biophys. J. 72(6), 2405–2412 (1997).
[Crossref] [PubMed]

J. E. Ferguson, S. M. Andrew, C. J. Jones, and P. J. August, “The Q-switched neodymium:YAG laser and tattoos: a microscopic analysis of laser-tattoo interactions,” Br. J. Dermatol. 137(3), 405–410 (1997).
[Crossref] [PubMed]

1991 (2)

C. R. Taylor, R. R. Anderson, R. W. Gange, N. A. Michaud, and T. J. Flotte, “Light and electron microscopic analysis of tattoos treated by Q-switched ruby laser,” J. Invest. Dermatol. 97(1), 131–136 (1991).
[Crossref] [PubMed]

D. Huang, E. A. Swanson, C. P. Lin, J. S. Schuman, W. G. Stinson, W. Chang, M. R. Hee, T. Flotte, K. Gregory, C. A. Puliafito, and et, “Optical coherence tomography,” Science 254(5035), 1178–1181 (1991).
[Crossref] [PubMed]

1990 (1)

W. Denk, J. H. Strickler, and W. W. Webb, “Two-photon laser scanning fluorescence microscopy,” Science 248(4951), 73–76 (1990).
[Crossref] [PubMed]

Ahn, G. O.

T. Wang, W. H. Jang, S. Lee, C. J. Yoon, J. H. Lee, B. Kim, S. Hwang, C. P. Hong, Y. Yoon, G. Lee, V. H. Le, S. Bok, G. O. Ahn, J. Lee, Y. S. Gho, E. Chung, S. Kim, M. H. Jang, S. J. Myung, M. J. Kim, P. T. So, and K. H. Kim, “Moxifloxacin: Clinically compatible contrast agent for multiphoton imaging,” Sci. Rep. 6(1), 27142 (2016).
[Crossref] [PubMed]

Akerman, L.

O. Reiter, L. Atzmony, L. Akerman, A. Levi, R. Kershenovich, M. Lapidoth, and D. Mimouni, “Picosecond lasers for tattoo removal: a systematic review,” Lasers Med. Sci. 31(7), 1397–1405 (2016).
[Crossref] [PubMed]

Anderson, R. R.

L. Izikson, W. Farinelli, F. Sakamoto, Z. Tannous, and R. R. Anderson, “Safety and effectiveness of black tattoo clearance in a pig model after a single treatment with a novel 758 nm 500 picosecond laser: a pilot study,” Lasers Surg. Med. 42(7), 640–646 (2010).
[Crossref] [PubMed]

M. Rajadhyaksha, S. González, J. M. Zavislan, R. R. Anderson, and R. H. Webb, “In vivo confocal scanning laser microscopy of human skin II: advances in instrumentation and comparison with histology,” J. Invest. Dermatol. 113(3), 293–303 (1999).
[Crossref] [PubMed]

V. Ross, G. Naseef, G. Lin, M. Kelly, N. Michaud, T. J. Flotte, J. Raythen, and R. R. Anderson, “Comparison of responses of tattoos to picosecond and nanosecond Q-switched neodymium: YAG lasers,” Arch. Dermatol. 134(2), 167–171 (1998).
[Crossref] [PubMed]

V. Ross, G. Naseef, G. Lin, M. Kelly, N. Michaud, T. J. Flotte, J. Raythen, and R. R. Anderson, “Comparison of responses of tattoos to picosecond and nanosecond Q-switched neodymium: YAG lasers,” Arch. Dermatol. 134(2), 167–171 (1998).
[Crossref] [PubMed]

C. R. Taylor, R. R. Anderson, R. W. Gange, N. A. Michaud, and T. J. Flotte, “Light and electron microscopic analysis of tattoos treated by Q-switched ruby laser,” J. Invest. Dermatol. 97(1), 131–136 (1991).
[Crossref] [PubMed]

Andrew, S. M.

J. E. Ferguson, S. M. Andrew, C. J. Jones, and P. J. August, “The Q-switched neodymium:YAG laser and tattoos: a microscopic analysis of laser-tattoo interactions,” Br. J. Dermatol. 137(3), 405–410 (1997).
[Crossref] [PubMed]

Arndt, K. A.

N. Saedi, A. Metelitsa, K. Petrell, K. A. Arndt, and J. S. Dover, “Treatment of tattoos with a picosecond alexandrite laser: a prospective trial,” Arch. Dermatol. 148(12), 1360–1363 (2012).
[Crossref] [PubMed]

Atzmony, L.

O. Reiter, L. Atzmony, L. Akerman, A. Levi, R. Kershenovich, M. Lapidoth, and D. Mimouni, “Picosecond lasers for tattoo removal: a systematic review,” Lasers Med. Sci. 31(7), 1397–1405 (2016).
[Crossref] [PubMed]

August, P. J.

J. E. Ferguson, S. M. Andrew, C. J. Jones, and P. J. August, “The Q-switched neodymium:YAG laser and tattoos: a microscopic analysis of laser-tattoo interactions,” Br. J. Dermatol. 137(3), 405–410 (1997).
[Crossref] [PubMed]

Aurangabadkar, S. J.

S. D. Shah and S. J. Aurangabadkar, “Newer trends in laser tattoo removal,” J. Cutan. Aesthet. Surg. 8(1), 25–29 (2015).
[Crossref] [PubMed]

S. J. Aurangabadkar, “Shifting paradigm in laser tattoo removal,” J. Cutan. Aesthet. Surg. 8(1), 3–4 (2015).
[Crossref] [PubMed]

Ayers, F. R.

F. R. Ayers, D. J. Cuccia, K. M. Kelly, and A. J. Durkin, “Wide-field spatial mapping of in vivo tattoo skin optical properties using modulated imaging,” Lasers Surg. Med. 41(6), 442–453 (2009).
[Crossref] [PubMed]

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Kim, M. J.

S. Lee, J. H. Lee, J. H. Park, Y. Yoon, W. K. Chung, H. Tchah, M. J. Kim, and K. H. Kim, “In vivo 3D measurement of moxifloxacin and gatifloxacin distributions in the mouse cornea using multiphoton microscopy,” Sci. Rep. 6(1), 25339 (2016).
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Supplementary Material (3)

NameDescription
» Visualization 1       TPM images before the laser treatment
» Visualization 2       TPM images 6 hours after the laser treatment (3 J/cm2)
» Visualization 3       TPM images 6 hours after the laser treatment (3 J/cm2)

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Figures (6)

Fig. 1
Fig. 1 Characterization of the fluorescent tattoo ink. (a) A FE-SEM image of the tattoo ink consisted of various sized particles. Scale bar indicates 2 μm. (b) Fluorescence emission spectra of the tattoo ink at various two-photon excitation wavelengths.
Fig. 2
Fig. 2 Representative TPM images of tattoo particles before and after the laser treatment. (a, b) TPM images of tattoo particles before and after the ns laser (4 J/cm2) treatment. (c, d) TPM images of tattoo particles before and after the ps laser (2 J/cm2) treatment. Image sets on the right shows zoomed images of selected particles marked with yellow-arrows 1 and 2, respectively. Scale bars in (a – d) and in zoomed images indicate 30 μm and 5 μm, respectively.
Fig. 3
Fig. 3 FE-SEM images of tattoo particle after laser treatments. (a, b) FE-SEM images after ns laser (4 J/cm2) treatment. (c, d) FE-SEM images after ps laser (2 J/cm2) treatment. (b) and (d) are zoomed images of black-dashed-box regions in (a) and (c), respectively. Yellow- and red-arrows indicate fragmented and intact tattoo particles, respectively. Scale bars indicate 3 μm.
Fig. 4
Fig. 4 TPM images of tattoo particles before and after the laser treatment at various laser fluences and the efficiency analysis of the ns and ps lasers in tattoo particle fragmentation. (a – c) ns laser treatment at 1, 2, 3, and 4 J/cm2 fluences. (d – f) ps laser at 0.5, 1, 1.5, and 2 J/cm2 fluences. Yellow- and red-arrowheads indicate intact small and large tattoo particles, respectively. (c, f) Quantitative treatment efficiency of the ns and ps lasers, respectively.
Fig. 5
Fig. 5 TPM images of tattoo particles in the mouse skin before and 6 hours after the treatment in vivo. (a, b) TPM images before and after the ns laser (2 J/cm2) treatment. (c, d) TPM images before and after the ps laser (1 J/cm2) treatment. Scale bars indicate 50 μm.
Fig. 6
Fig. 6 Time-lapse TPM images of the mouse skin dermis before and after the ns laser treatment. (a – c) TPM images before the laser treatment (Visualization 1). (d – h) TPM images 6 hours after the laser treatment (3 J/cm2) (Visualization 2, Visualization 3). Red and green colors in TPM images indicate tattoo and moxifloxacin fluorescence, respectively. Zoomed time-lapse TPM images show both tattoo particles and cells at different time points. Time stamps in the zoomed images depict relative time points with respect to the starting time of the time-lapse imaging. Scale bars indicate 100 μm.

Tables (1)

Tables Icon

Table 1 Laser specifications and experimental conditions of ns and ps pulse lasers

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