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Detection of Micrococcus luteus biofilm formation in microfluidic environments by pH measurement using an ion-sensitive field-effect transistor (CAT#: STEM-PPA-0004-LJX)

Introduction

Biofilm formation in microfluidic channels is difficult to detect because sampling volumes are too small for conventional turbidity measurements. To detect biofilm formation, the service uses an ion-sensitive field-effect transistor (ISFET) measurement system to measure pH changes in small volumes of bacterial suspension. Cells of Micrococcus luteus (M. luteus) were cultured in polystyrene (PS) microtubes and polymethylmethacrylate (PMMA)-based microfluidic channels laminated with polyvinylidene chloride. In microtubes, concentrations of bacteria and pH in the suspension were analyzed by measuring turbidity and using an ISFET sensor, respectively. In microfluidic channels containing 20 μL of bacterial suspension, we measured pH changes using the ISFET sensor and monitored biofilm formation using a microscope.




Principle

The ion-sensitive field-effect transistor method (ISEFT) uses voltage source and drain (source and channel) transistors separated by insulators. Insulators (gates) are made of metal oxides. Hydrogen ions in the medium can accumulate on it, as on a glass electrode, and the positive charge generated by its outer layer will form a "mirror symmetry" in the inner layer of the gate, resulting in a negative charge, so that the gate has electrical conductivity. The lower the pH of the solution, the more hydrogen ions are concentrated on the gate, and the more current flowing between the source and drain can be measured.

Applications

For determining the approximate PH value of solutions in food production, wastewater treatment, pharmaceutical and chemical industries

Procedure

1. Select the "pH" file of the instrument, immerse the cleaned electrode into the standard pH buffer solution to be measured, press the measurement button, turn the positioning adjustment knob, so that the pH value displayed by the instrument is stable at the pH value of the standard buffer solution
2. Release the measuring button, take out the electrode, rinse with distilled water several times, and carefully blot the solution on the electrode with filter paper
3. Place the electrode in the liquid to be tested, press the measurement button, read the stable pH value, and record
4. Release the measurement button, remove the electrode, clean according to step 2, and continue to measure the next sample solution
5. After measuring, clean the electrode and soak the glass electrode in distilled water

Materials

• Sample Type:
Cells of Micrococcus luteus

Notes

1. If you need to measure pH with high accuracy, in order to avoid the effect of CO2 in the air, especially the pH of alkaline solutions, the exposure time to the air should be as short as possible, and the reading should be as fast as possible.
2. In order to ensure the accuracy of the results, the electrodes need to be cleaned in time before and after each measurement.