Nanosensor Quantifying Lysosomal Glycosidase Secretion from Single Living Cells.

Area of Science:

• Immunology
• Cell Biology
• Biomedical Engineering

Background:

• Secretory lysosomes are vital for immune cell function, regulating both degradation and secretion.
• Disruptions in lysosomal secretion can cause immune imbalances and health issues.
• Existing methods like immunostaining lack real-time, quantitative analysis and compromise cell viability.

Purpose of the Study:

• To develop an innovative method for real-time, quantitative monitoring of lysosomal enzyme secretion from single living immune cells.
• To overcome limitations of current immunostaining techniques.
• To investigate the dynamics of secretory lysosomes in maintaining immune homeostasis.

Main Methods:

• Combined artificial substrates with hybrid nanowire-nanopipette sensors.
• Induced glycosidase secretion in macrophages via frustrated phagocytosis.
• Quantified secreted glycosidases by detecting electroactive aminophenol released from substrate hydrolysis using nanowire electrodes.

Main Results:

• Successfully quantified lysosomal glycosidase secretion from individual living macrophages.
• Demonstrated a novel approach for real-time, quantitative assessment of enzyme secretion.
• Provided insights into the dynamic role of secretory lysosomes in immune regulation.

Conclusions:

• Developed a versatile methodology for monitoring lysosomal enzyme secretion in single living macrophages.
• Highlighted the importance of secretory lysosomes in maintaining immune homeostasis.
• Established a new tool for studying immune cell function and related diseases.