Intracellular labeling method for chip-based capillary electrophoresis fluorimetric single cell analysis using...

Area of Science:

• Biotechnology
• Analytical Chemistry
• Cell Biology

Background:

• Liposome-mediated intracellular delivery is crucial for single-cell analysis.
• Existing methods often struggle with efficiency and cell viability.
• Novel approaches are needed for precise intracellular labeling.

Purpose of the Study:

• To develop a liposome-mediated intracellular derivatization method for single-cell analysis.
• To enable the delivery of non-permeant fluorescent dyes into living cells.
• To combine this method with chip-based capillary electrophoresis (CE) and laser-induced fluorescence (LIF) detection.

Main Methods:

• Liposomes (100 nm diameter) were prepared from phosphatidylcholine using an ultrasonic method to encapsulate fluorescent dyes.
• Encapsulation yield (46+/-5%) and vesicle density (8.8 x 10^14/mL) were determined.
• Cell incubation time, liposome density, and dye concentration influenced dye uptake.

Main Results:

• Liposomes successfully delivered cell membrane non-permeable fluorescent dyes into living cells.
• Cell viability was maintained throughout the delivery process.
• Liposome-membrane fusion within cells released dyes, enabling intracellular labeling and analysis via chip-based CE-LIF.

Conclusions:

• Liposome-mediated intracellular derivatization is an effective strategy for single-cell analysis.
• This technique allows for the introduction of otherwise impermeable fluorescent dyes into cells.
• The method is compatible with sensitive detection techniques like CE-LIF, preserving cell integrity.