Live Imaging of Fetal Intra-abdominal Organs Using Two-Photon Laser-Scanning Microscopy

  • Yuhki Koike
  • Bo Li
  • Yong Chen
  • Hiromu Miyake
  • Carol Lee
  • Lijun Chi
  • Richard Wu
  • Mikihiro Inoue
  • Keiichi Uchida
  • Masato Kusunoki
  • Paul Delgado-Olguin
  • Agostino Pierro
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1752)

Abstract

The processes by which the intra-abdominal organ circulatory system develops in the embryo and during organogenesis are unclear. Previous studies have used fixed tissues to study the development of abdominal organ vasculature in the embryo; however, the intravital circulation of intra-abdominal organs in rodent fetal development has not been studied. This protocol describes a system that uses two-photon laser-scanning microscopy (TPLSM) for real-time observation and quantification of normal and pathologic live fetal intra-abdominal dynamics while the fetus is still connected to the mother via the umbilical cord.

Key words

Two-photon laser scanning microscopy (TPLSM) Intravital imaging Fetus 

References

  1. 1.
    Koike Y, Tanaka K, Kobayashi M, Toiyama Y, Inoue Y, Mohri Y et al (2015) Dynamic pathology for leukocyte-platelet formation in sepsis model. J Surg Res 195(1):188–195CrossRefPubMedGoogle Scholar
  2. 2.
    Tanaka K, Koike Y, Shimura T, Okigami M, Ide S, Toiyama Y et al (2014) In vivo characterization of neutrophil extracellular traps in various organs of a murine sepsis model. PLoS One 9(11):e111888CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Mizuno R, Kamioka Y, Kabashima K, Imajo M, Sumiyama K, Nakasho E et al (2014) In vivo imaging reveals PKA regulation of ERK activity during neutrophil recruitment to inflamed intestines. J Exp Med 211(6):1123–1136CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Koike Y, Uchida K, Tanaka K, Ide S, Otake K, Okita Y et al (2014) Dynamic pathology for circulating free DNA in a dextran sodium sulfate colitis mouse model. Pediatr Surg Int 30(12):1199–1206CrossRefPubMedGoogle Scholar
  5. 5.
    Dunn KW, Sandoval RM, Molitoris BA (2003) Intravital imaging of the kidney using multiparameter multiphoton microscopy. Nephron Exp Nephrol 94(1):e7–11CrossRefPubMedGoogle Scholar
  6. 6.
    Zenclussen AC, Olivieri DN, Dustin ML, Tadokoro CE (2013) In vivo multiphoton microscopy technique to reveal the physiology of the mouse uterus. Am J Reprod Immunol 69(3):281–289CrossRefPubMedGoogle Scholar
  7. 7.
    Zenclussen AC, Olivieri DN, Dustin ML, Tadokoro CE (2012) In vivo multiphoton microscopy technique to reveal the physiology of the mouse placenta. Am J Reprod Immunol 68(3):271–278CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Yuhki Koike
    • 1
    • 2
  • Bo Li
    • 1
  • Yong Chen
    • 1
  • Hiromu Miyake
    • 1
  • Carol Lee
    • 1
  • Lijun Chi
    • 3
    • 4
  • Richard Wu
    • 5
  • Mikihiro Inoue
    • 2
  • Keiichi Uchida
    • 2
  • Masato Kusunoki
    • 2
  • Paul Delgado-Olguin
    • 7
    • 8
    • 9
  • Agostino Pierro
    • 1
    • 6
  1. 1.Translational Medicine Program, Division of General and Thoracic SurgeryThe Hospital for Sick ChildrenTorontoCanada
  2. 2.Department of Gastrointestinal and Pediatric SurgeryMie University Graduate School of MedicineTsuJapan
  3. 3.Translational MedicineThe Hospital for Sick ChildrenTorontoCanada
  4. 4.Department of Molecular GeneticsUniversity of TorontoTorontoCanada
  5. 5.Department of Gastroenterology, Hepatology and NutritionThe Hospital for Sick ChildrenTorontoCanada
  6. 6.The Hospital for Sick ChildrenUniversity of TorontoTorontoCanada
  7. 7.Translational MedicineThe Hospital for Sick ChildrenTorontoCanada
  8. 8.Department of Molecular GeneticsUniversity of TorontoTorontoCanada
  9. 9.Heart & Stroke Richard Lewar Centres of Excellence in Cardiovascular ResearchTorontoCanada

Personalised recommendations