Skip to main content

Microscopy Tools for Quantifying Developmental Dynamics in Xenopus Embryos

  • Protocol
  • First Online:
Xenopus Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 917))

Abstract

Early Xenopus embryos, and embryonic tissues isolated from them, are excellent model systems to study morphogenesis. Cells migrate, change shape, and differentiate to form new tissues as embryos mature and recapitulate those same processes in tissue isolates. Both large-scale and small-scale cell and tissue movements can be visualized with a range of microscopy techniques. Furthermore, protein dynamics, fine-scale cell movements, and changes in cell morphology can be observed simultaneously as multicellular structures are sculpted. We provide an overview of complementary methods for visualizing macroscopic tissue movements, cell shape changes, and subcellular protein dynamics. Time-lapse imaging followed by quantitative image analysis aims to provide answers to some of the long-standing questions in developmental biology: How do tissues form? How do cells acquire specific shapes? How do proteins localize to specific positions? To address these questions we suggest strategies (1) to visualize whole embryos and tissue isolates using stereoscopes and epifluorescence imaging techniques, and (2) to visualize cell shapes and protein expression using high-resolution live imaging using confocal microscopy. These imaging approaches along with simple image analysis tools provide us with ways to understand the complex biology underlying morphogenesis.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 99.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 129.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 199.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Sive HL, Grainger RM, Harland RM (eds) (2000) Early development of Xenopus laevis: a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, p 338

    Google Scholar 

  2. Kim HY, Davidson LA (2010) Methods to investigate molecular mechanisms and cellular mechanics responsible for morphogenesis in Xenopus laevis embryos. In: Wong R, Sharpe J (eds) Imaging in developmental biology: a laboratory manual. Cold Spring Harbor Laboratory Press, New York

    Google Scholar 

  3. Arganda-Carreras I et al (2006) Consistent and elastic registration of histological sections using vector-spline regularization. Computer Vision Approaches to Medical Image Analysis, ser. Lecture Notes in Computer Science, vol 4241. Springer, Berlin, p 85–95

    Google Scholar 

  4. Sorzano CO, Thevenaz P, Unser M (2005) Elastic registration of biological images using vector-spline regularization. IEEE Trans Biomed Eng 52(4):652–663

    Article  PubMed  Google Scholar 

  5. Lane MC, Davidson L, Sheets MD (2004) BMP antagonism by Spemann’s organizer regulates rostral-caudal fate of mesoderm. Dev Biol 275(2):356–374

    Article  CAS  Google Scholar 

  6. Tsien RY, Waggoner AF (1995) Fluorophores for confocal microscopy. In: Pawley JB (ed) Handbook of biological confocal microscopy, 2nd edn. Plenum Press, New York, pp 267–280

    Google Scholar 

  7. Joshi SD, Davidson LA (2010) Live-cell imaging and quantitative analysis of embryonic epithelial cells in Xenopus laevis. J Vis Exp 23(39):pii: 1949

    Google Scholar 

  8. Joshi SD, von Dassow M, Davidson LA (2010) Experimental control of excitable embryonic tissues: three stimuli induce rapid epithelial contraction. Exp Cell Res 316(1):103–114

    Article  PubMed  CAS  Google Scholar 

  9. Kim HY, Davidson LA (2011) Punctuated actin contractions during convergent extension and their permissive regulation by the non-canonical Wnt-signaling pathway. J Cell Sci 124(4):635–646

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgements

We would like to thank past and present members of the Davidson, Keller, and DeSimone Labs for their care and patience in helping to develop these methods through trial and mostly error. In addition, we would like to thank Lin Zhang for her assistance. This work was made possible through grants from the NIH (R01 HD044750, R21 ES019259) and from a CAREER award from the NSF (IOS-0845775).

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Lance A. Davidson .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2012 Springer Science+Business Media, LLC

About this protocol

Cite this protocol

Joshi, S.D., Kim, H.Y., Davidson, L.A. (2012). Microscopy Tools for Quantifying Developmental Dynamics in Xenopus Embryos. In: HOPPLER, S., Vize, P. (eds) Xenopus Protocols. Methods in Molecular Biology, vol 917. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-992-1_27

Download citation

  • DOI: https://doi.org/10.1007/978-1-61779-992-1_27

  • Published:

  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-61779-991-4

  • Online ISBN: 978-1-61779-992-1

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics