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Related Experiment Video

Updated: May 3, 2026

Lineage Tracing and Clonal Analysis in Developing Cerebral Cortex Using Mosaic Analysis with Double Markers MADM
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Mosaic analysis with double markers (MADM) in mice.

J Sebastian Espinosa, Joy S Tea, Liqun Luo

    Cold Spring Harbor Protocols
    |February 5, 2014
    PubMed
    Summary

    Mosaic analysis with double markers (MADM) enables researchers to visualize and manipulate individual neurons. This genetic method is crucial for understanding neural circuits, lineage analysis, and gene function in vivo.

    Area of Science:

    • Neuroscience
    • Genetics
    • Molecular Biology

    Background:

    • The human brain has over 100 billion neurons with complex connectivity.
    • Visualizing individual neurons is essential for understanding brain complexity.
    • Existing methods may not allow simultaneous labeling and gene manipulation.

    Purpose of the Study:

    • To provide a standard procedure for Mosaic Analysis with Double Markers (MADM).
    • To demonstrate MADM's utility in lineage analysis, neural circuit tracing, and gene function studies.
    • To detail protocols and troubleshooting for MADM application.

    Main Methods:

    • Utilizing site-specific recombinases for chimeric fluorescent protein reconstitution.
    • Employing MADM for simultaneous labeling and gene knockout in somatic cell clones or single cells.

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  • Using ROSA26-MADM as a specific example with available reagents.

    • Main Results:

    • Detailed protocols for MADM are presented.
    • Troubleshooting guides are included for practical application.
    • Applications in neural circuit tracing and studying neuronal morphogenesis are demonstrated.

    Conclusions:

    • MADM is a powerful genetic method for individual neuron analysis.
    • The technique facilitates in vivo lineage tracing and gene function studies.
    • MADM offers a versatile platform for advancing neuroscience research.