Protocol to quantify total intracellular magnesium in small samples using a fluorescent plate reader and fluorescent...

Area of Science:

• Cellular Biology
• Biochemistry
• Analytical Chemistry

Background:

• Accurate measurement of intracellular magnesium (Mg) is crucial for understanding cellular function.
• Traditional methods for Mg quantification are often unsuitable for samples with limited cell numbers.

Purpose of the Study:

• To present a reliable protocol for quantifying total intracellular magnesium (Mg) in samples containing minimal cell quantities.
• To enable Mg assessment using a fluorescent plate reader and a specific fluorescent dye.

Main Methods:

• Development and validation of a protocol for total intracellular Mg quantification.
• Utilizing the fluorescent dye diaza-18-crown-6-hydroxyquinoline-5 (DCHQ5).
• Employing a fluorescent plate reader for sample analysis with as few as several thousand cells.

Main Results:

• Successful quantification of total intracellular Mg in samples with minimal cell numbers (thousands of cells).
• The protocol is detailed, covering reagent setup, sample preparation, assay execution, and data analysis.
• Validation of the protocol across different cell lines, supported by existing literature data.

Conclusions:

• The presented protocol offers a sensitive and accessible method for intracellular Mg measurement.
• This technique significantly expands the feasibility of Mg assessment in low-cell-number biological samples.
• Facilitates further research into the role of magnesium in cellular processes.