A novel dual-release scaffold for fluorescent labels improves cyclic immunofluorescence

Area of Science:

• Cell Biology
• Biotechnology
• Immunology

Background:

• Cyclic immunofluorescence generates high-content imaging data for cell biology and therapeutics.
• Fluorescent labels are crucial for immunofluorescence quality and multiplexing capabilities.
• Current labels face limitations in signal erasure efficiency, restricting the number of staining cycles.

Purpose of the Study:

• To develop a novel fluorescent labelling strategy for enhanced cyclic immunofluorescence.
• To create labels with improved brightness and efficient signal erasure.
• To advance multiplexed cyclic immunofluorescence for deeper biological insights.

Main Methods:

• Antibodies were conjugated to a novel scaffold for enzymatic fluorophore cleavage.
• The scaffold comprised dextran decorated with single-stranded DNA (ssDNA).
• Dual enzymatic cleavage mechanisms (DNase and dextranase) were employed for signal erasure.

Main Results:

• The developed fluorescent labels demonstrated specific staining and high brightness.
• Labels performed effectively in flow cytometry and fluorescence microscopy.
• Dual enzymatic degradation significantly improved signal erasure from labelled epitopes.

Conclusions:

• The novel dual-release fluorescent labels enhance cyclic immunofluorescence.
• These labels offer high brightness and efficient, specific signal erasure.
• The strategy is expected to advance multiplexed cyclic immunofluorescence and cell biology research.