In this method, embryonic cardiac tissues are surgically microdissected, dissociated, fluorescently labeled, and implanted into host embryonic tissues. This provides a platform for studying the individual or tissue level developmental organization under ectopic hemodynamic conditions, and/or altered paracrine/juxtacrine environments.
Total time
~4–6 hours per embryo pair (including tissue isolation, dissociation, injection, and live imaging)
Model organism
Avian embryo (chicken)
Steps
1
Prepare microinjection pipettes for cell delivery
Fabricate and calibrate glass micropipettes suitable for precise injection of dissociated cardiomyocytes. Verify pipette diameter and flow characteristics before use.
▶ 00:49
2
Prepare host embryo for cell implantation
Access and position the embryonic heart in the host avian embryo under stereomicroscopy to establish the surgical target site for injection.
▶ 01:53
3
Isolate donor embryonic cardiac tissue
Microdissect intact embryonic cardiac tissue from the donor embryo under magnification and collect in appropriate dissociation medium.
▶ 03:34
4
Dissociate and label donor cardiomyocytes
Enzymatically digest isolated donor cardiac tissue to obtain single cardiomyocytes, then apply fluorescent labeling for downstream visualization.
▶ 04:44
5
Inject labeled cells into host embryonic heart
Using the prepared microinjection pipette, deliver dissociated and labeled donor cardiomyocytes into the target site within the host embryonic heart tissue.
▶ 05:50
6
Image implanted myocytes in recipient heart
Acquire fluorescence and/or morphological images of donor-derived myocytes integrated within the host heart to assess engraftment and organization.
▶ 07:37