Display options
Cite
Song E, Seo H, Choe K, et al. Optical clearing based cellular-level 3D visualization of intact lymph node cortex. Biomed Opt Express. 2015;6(10):4154-64doi: 10.1364/BOE.6.004154.
Song, E., Seo, H., Choe, K., Hwang, Y., Ahn, J., Ahn, S., & Kim, P. (2015). Optical clearing based cellular-level 3D visualization of intact lymph node cortex. Biomedical optics express, 6(10), 4154-64. https://doi.org/10.1364/BOE.6.004154
Song, Eunjoo, et al. "Optical clearing based cellular-level 3D visualization of intact lymph node cortex." Biomedical optics express vol. 6,10 (2015): 4154-64. doi: https://doi.org/10.1364/BOE.6.004154
Song E, Seo H, Choe K, Hwang Y, Ahn J, Ahn S, Kim P. Optical clearing based cellular-level 3D visualization of intact lymph node cortex. Biomed Opt Express. 2015 Sep 28;6(10):4154-64. doi: 10.1364/BOE.6.004154. eCollection 2015 Oct 01. PMID: 26504662; PMCID: PMC4605071.
Copy Download .nbib Format: NLM
Share it on

Biomed Opt Express. 2015 Sep 28;6(10):4154-64. doi: 10.1364/BOE.6.004154. eCollection 2015 Oct 01.

Optical clearing based cellular-level 3D visualization of intact lymph node cortex.

Biomedical optics express

Eunjoo Song, Howon Seo, Kibaek Choe, Yoonha Hwang, Jinhyo Ahn, Soyeon Ahn, Pilhan Kim

Affiliations

  1. Graduate School of Nanoscience and Technology, Korea Advanced Institute of Science and Technology (KAIST), Daejeon, 305-701, South Korea.

PMID: 26504662 PMCID: PMC4605071 DOI: 10.1364/BOE.6.004154

Abstract

Lymph node (LN) is an important immune organ that controls adaptive immune responses against foreign pathogens and abnormal cells. To facilitate efficient immune function, LN has highly organized 3D cellular structures, vascular and lymphatic system. Unfortunately, conventional histological analysis relying on thin-sliced tissue has limitations in 3D cellular analysis due to structural disruption and tissue loss in the processes of fixation and tissue slicing. Optical sectioning confocal microscopy has been utilized to analyze 3D structure of intact LN tissue without physical tissue slicing. However, light scattering within biological tissues limits the imaging depth only to superficial portion of LN cortex. Recently, optical clearing techniques have shown enhancement of imaging depth in various biological tissues, but their efficacy for LN are remained to be investigated. In this work, we established optical clearing procedure for LN and achieved 3D volumetric visualization of the whole cortex of LN. More than 4 times improvement in imaging depth was confirmed by using LN obtained from H2B-GFP/actin-DsRed double reporter transgenic mouse. With adoptive transfer of GFP expressing B cells and DsRed expressing T cells and fluorescent vascular labeling by anti-CD31 and anti-LYVE-1 antibody conjugates, we successfully visualized major cellular-level structures such as T-cell zone, B-cell follicle and germinal center. Further, we visualized the GFP expressing metastatic melanoma cell colony, vasculature and lymphatic vessels in the LN cortex.

Keywords: (170.1790) Confocal microscopy; (170.3660) Light propagation in tissues; (170.3880) Medical and biological imaging; (170.6935) Tissue characterization

References

  1. Cancer Res. 2011 Feb 15;71(4):1214-8 - PubMed
  2. Nat Methods. 2007 Apr;4(4):331-6 - PubMed
  3. Proc Natl Acad Sci U S A. 2005 Nov 8;102(45):16315-20 - PubMed
  4. PLoS One. 2012;7(3):e33916 - PubMed
  5. Blood. 2011 Jan 13;117(2):563-74 - PubMed
  6. Opt Express. 2014 May 19;22(10):11465-75 - PubMed
  7. Biomed Opt Express. 2013 Sep 05;4(10):1978-90 - PubMed
  8. Microsc Res Tech. 2008 Jul;71(7):510-6 - PubMed
  9. Am J Pathol. 2007 Feb;170(2):774-86 - PubMed
  10. J Exp Med. 2014 Dec 15;211(13):2497-505 - PubMed
  11. Cell. 2014 Apr 24;157(3):726-39 - PubMed
  12. Immunity. 2011 Jan 28;34(1):96-107 - PubMed
  13. Science. 2002 Jun 7;296(5574):1869-73 - PubMed
  14. Nat Med. 2012 Jun;18(6):883-91 - PubMed
  15. J Clin Pathol. 2004 Jan;57(1):64-7 - PubMed
  16. Curr Opin Cell Biol. 2015 Oct;36:1-6 - PubMed
  17. Cell. 2015 Jul 16;162(2):246-57 - PubMed
  18. PLoS One. 2013;8(1):e53753 - PubMed
  19. Photochem Photobiol. 2011 May-Jun;87(3):734-40 - PubMed
  20. Biomed Opt Express. 2013 Dec 04;5(1):69-77 - PubMed
  21. Blood. 2007 Feb 1;109(3):1010-7 - PubMed
  22. J Transl Med. 2012 Oct 04;10:206 - PubMed
  23. Science. 2006 Jun 16;312(5780):1672-6 - PubMed
  24. J Exp Med. 2012 Apr 9;209(4):653-60 - PubMed
  25. Nat Med. 2012 Nov;18(11):1614-6 - PubMed
  26. Opt Express. 2012 Aug 27;20(18):20582-98 - PubMed
  27. Cancer Res. 2011 Jun 1;71(11):3792-801 - PubMed
  28. J Exp Med. 2013 Jul 29;210(8):1509-28 - PubMed
  29. J Immunol. 2012 Apr 15;188(8):4065-80 - PubMed
  30. Biomed Opt Express. 2015 May 19;6(6):2158-67 - PubMed
  31. Nat Immunol. 2007 Sep;8(9):992-1000 - PubMed
  32. Cancer Res. 2006 Nov 1;66(21):10365-76 - PubMed
  33. Trends Immunol. 2012 Jun;33(6):306-14 - PubMed
  34. Trends Immunol. 2012 Jun;33(6):271-80 - PubMed
  35. Biomed Opt Express. 2013 May 30;4(6):967-77 - PubMed
  36. J Biomed Opt. 2010 Jul-Aug;15(4):046018 - PubMed
  37. Gastroenterology. 2009 Aug;137(2):453-65 - PubMed
  38. Science. 2010 Jan 15;327(5963):291-5 - PubMed
  39. Biomed Opt Express. 2013 Oct 17;4(11):2518-26 - PubMed
  40. J Biomed Opt. 2013 Mar;18(3):036005 - PubMed

Publication Types