Combining Fluorescent In Situ Hybridization with Immunofluorescence and Lectin Staining in Planarians.

Area of Science:

• Molecular Biology
• Developmental Biology
• Cell Biology

Background:

• Simultaneous visualization of genetic expression and tissue composition in Schmidtea mediterranea is crucial for understanding its biology.
• Fluorescent in situ hybridization (FISH) and immunofluorescence (IF) are key techniques for such analyses.
• Expanding detection capabilities beyond FISH and IF is highly desirable.

Purpose of the Study:

• To develop a novel combined protocol for FISH and IF in Schmidtea mediterranea.
• To integrate fluorescent-conjugated lectin staining into the combined FISH-IF protocol.
• To introduce an improved lectin fixation method for enhanced signal detection.

Main Methods:

• A novel protocol combining FISH and IF was developed.
• Fluorescent-conjugated lectin staining was incorporated into the FISH-IF workflow.
• A new lectin fixation protocol was optimized for improved signal.

Main Results:

• The combined FISH-IF protocol allows simultaneous visualization of molecular markers and tissue structures.
• Lectin staining successfully broadened the scope of detectable tissue components.
• The novel lectin fixation protocol significantly enhanced signal intensity, facilitating single-cell resolution.

Conclusions:

• The developed combined FISH-IF and lectin staining protocol offers a powerful tool for multi-modal analysis in Schmidtea mediterranea.
• The enhanced lectin fixation method improves signal-to-noise ratio for high-resolution imaging.
• This integrated approach advances the study of gene expression and tissue dynamics in planarian flatworms.