A modified single-cell electroporation method for molecule delivery into a motile protist, Euglena gracilis

Area of Science:

• Cell Biology
• Microbiology
• Biotechnology

Background:

• Single-cell transfection is crucial for studying individual microscopic organisms.
• Analyzing motile protists like Euglena gracilis is challenging due to difficulties in tracking.
• Existing bulk population labeling methods hinder individual cell analysis.

Purpose of the Study:

• To modify single-cell electroporation for efficient molecule delivery into Euglena gracilis.
• To improve transfection efficiency and cell viability in motile microalgae.
• To enable precise molecular and cellular behavior studies in individual protists.

Main Methods:

• Modified single-cell electroporation technique using negative pressure via micropipette.
• Delivery of fluorescent markers (GFP, Alexa Fluor 488) and ECHO RNA probes.
• Electroporation applied to individual, living Euglena gracilis cells.

Main Results:

• Achieved high transfection efficiency and post-electroporation viability in Euglena gracilis.
• Successfully introduced various molecules, including fluorescent probes and RNA probes.
• Demonstrated imaging of endogenous mRNA in living Euglena gracilis without affecting physiological functions.

Conclusions:

• The modified single-cell electroporation technique is effective for delivering diverse molecules into individual motile protists.
• This method facilitates long-term observation of cellular functions like swimming and division.
• Enables versatile functional molecule delivery for studying individual motile microalgae.