A microchamber array for single cell isolation and analysis of intracellular biomolecules

Area of Science:

• Biotechnology
• Cell Biology
• Analytical Chemistry

Background:

• Analyzing intracellular biomolecules in single cells is crucial for understanding cellular heterogeneity and function.
• Existing methods often require large cell numbers or lack precise control over sequential cell treatments.

Purpose of the Study:

• To develop and validate a novel microfluidic device for high-throughput single-cell analysis.
• To demonstrate the device's capability in monitoring intracellular cofactors and enzyme activity under various conditions.

Main Methods:

• A microfluidic device with an array of individually addressable and sealable microchambers was designed.
• Cells were trapped, sequentially treated (incubation, washing, labeling, lysis), and their intracellular contents analyzed.
• Levels of nicotinamide adenine dinucleotide phosphate (NADPH), nicotinamide adenine dinucleotide (NADH), and glucose-6-phosphate dehydrogenase were quantified.

Main Results:

• The device successfully isolated and processed single mammalian cells for intracellular analysis.
• Changes in NADPH and NADH levels were monitored in cells under oxidative stress.
• The toxic effect of camptothecin on glucose-6-phosphate dehydrogenase activity was successfully determined.

Conclusions:

• The microfluidic platform provides a versatile tool for analyzing intracellular biomolecules in single cells.
• It enables precise control over sequential cell treatments and sensitive detection of metabolites and enzyme activities.
• This technology has broad applications in cell biology, drug screening, and disease research.