Related Experiment Video
Updated: Jun 4, 2026
Analyzing In Vivo Cell Migration using Cell Transplantations and Time-lapse Imaging in Zebrafish Embryos
Published on: April 29, 2016
Analysis of cell movement in amphimedon embryos by injection of fluorescent tracers
Maja Adamska1, Bernard M Degnan
1School of Integrative Biology, University of Queensland, Brisbane QLD 4072, Australia.
Abstract:
INTRODUCTIONAlthough attempts to culture prepigmentation-stage embryos (i.e., blastulas and early gastrulas) outside of brood chambers have so far been unsuccessful in Amphimedon, it is possible to manipulate embryos within the brood chamber and follow their development under laboratory conditions. This protocol describes microinjection of lipophilic tracers such as 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate (DiI) into embryos embedded in their native brood chamber. DiI does not appear to perturb embryonic development and is relatively resistant to photobleaching. As long as care is taken not to damage the fragile embryos during observation and photography, the same embryo can be photographed multiple times, permitting its development to be tracked (up to 4 wk) from early cleavage stages to hatching of free-swimming parenchymella larvae. The embryos or larvae also can be fixed without loss of fluorescence. This method also can be used to deliver other types of solutions to embryos or individual cells of early embryos.
Insights
This study introduces a method for tracking Amphimedon embryonic development within brood chambers using lipophilic tracers like DiI. This technique allows for long-term observation and fixation of embryos without perturbing their development.
Area of Science:
- Marine Biology
- Developmental Biology
- Cell Biology
Background:
- Culturing early-stage Amphimedon embryos (blastulas and gastrulas) outside brood chambers has proven unsuccessful.
- Previous methods lacked the ability to track individual embryo development over extended periods in their native environment.
Purpose of the Study:
- To develop a protocol for manipulating and observing Amphimedon embryos within their native brood chambers.
- To enable long-term tracking of embryonic development from cleavage to larval stages.
- To facilitate the delivery of substances to early embryos or individual cells.
Main Methods:
- Microinjection of lipophilic tracers, specifically 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate (DiI), into Amphimedon embryos within their brood chambers.
- Utilizing DiI, which is resistant to photobleaching and does not appear to disrupt development.
- Repeated observation and photography of the same embryo over a period of up to 4 weeks.
- Fixation of embryos or larvae post-observation while preserving fluorescence.
Main Results:
- The developed protocol allows for successful microinjection and tracking of Amphimedon embryonic development within brood chambers.
- DiI labeling did not perturb embryonic development, enabling observation from early cleavage to the hatching of free-swimming parenchymella larvae.
- The same embryo could be repeatedly photographed, and embryos/larvae could be fixed without fluorescence loss.
Conclusions:
- This method provides a novel approach for studying early embryonic development in Amphimedon under laboratory conditions.
- The technique is valuable for tracking developmental processes and allows for subsequent molecular analysis.
- The protocol's adaptability extends to delivering various solutions to embryos or single cells, offering broad applications in developmental studies.
