Measurement of Intracellular pH in Plant Cells

Intracellular pH (pH) is an important physiological determinant of enzyme activity and cellular function, and all proteins depend on a tightly regulated pH to maintain their structure and function. Intracellular pH, as an essential regulator of cellular physiological activities, not only regulates enzyme activities and some critical metabolic processes, but also many physiological activities such as ATP synthesis, DNA replication, RNA and protein synthesis, and cell growth are regulated by intracellular pH. More and more experiments have shown that changes in intracellular pH act as a second messenger in physiological activities such as apical growth, root gravitropism, plant defense, and phytohormone response. The pH value in the cytoplasm of most plants is close to neutral, but there will be some changes under special physiological conditions or external environmental factors. The timely intracellular pH regulation of plant cells is an important feature of all living cells, and intracellular pH is also an important parameter in the current study of cell function. Therefore, accurate, dynamic, and non-destructive observation of intracellular pH is important for the in-depth analysis of the relationship between cell metabolism and intracellular pH and the related mechanism of action.

Lifeasible uses suitable methods for measuring intracellular pH according to different experimental materials before measurement. Currently, we mainly utilize nuclear magnetic resonance (NMR), microelectrode, and fluorescent dye methods to determine intracellular pH in plants.

What do we offer?

• Nuclear Magnetic Resonance (NMR). This technique does not require an exogenous molecular probe, is non-invasive to the cell, and avoids the physiological and metabolic changes within the cell caused by various types of damage or disruption and the resulting inaccuracies in the measurement results. The method allows for simultaneous determination of cytoplasmic and vesicular pH, and it is easy to observe the heterogeneity of pH in tissue cells. In addition, this method can not only understand the intracellular pH regulation mechanism. It can also obtain information on a variety of phosphorus-containing compounds such as intracellular phosphate monoesters, phosphate diesters, ATP, bisphosphate diesters, phosphocreatine, and inorganic phosphorus (Pi), and is therefore of great value for application. The main limitations of this method are the low sensitivity, the need for a large amount of biological material (1-5 g fresh weight) or a high concentration of cells for the determination, and the fact that the study of pH changes in cell populations or organ parts is more appropriate with the NMR technique.
• Fluorescent dyes. We use pH-sensitive fluorescent dyes that have been effectively used to determine intracellular pH. The fluorescence intensity of pH fluorescent dyes is dependent on intracellular pH, and the fluorescence intensity can be measured using specific instruments such as a laser confocal microscope or a flow cytometer, and then, based on the in vitro or in vivo calibration curves, the intracellular pH of the cell being measured can be derived. The measurement requires selecting the appropriate concentration of fluorescent dye, the temperature during staining, and the staining time for the material. The main advantages of this method are the relative simplicity of operation, the ability to monitor intracellular pH changes in real-time, the applicability to cells of various sizes, the lack of limitation of cell concentration, the good reproducibility, and the ability to determine the pH of multiple intracellular sites at the same time.

In addition, we can also use the photoelectric method for the determination of intracellular pH in plant cells, using photodetectors to measure the photoelectric signals inside the cell and inferring the changes in intracellular pH through the changes in the signals, which is a more accurate method.

The report we delivered at the end of the project is as follows

1. Experimental steps
2. Relevant parameters
3. Pictures
4. raw data
5. Information on plant pH content

Advantages of our services

Lifeasible has many years of experience in plant cellular measurement and functional analysis, and we always follow the development of advanced technology, using various advanced technology methods, which can determine plant pH value efficiently and accurately. In addition, we also provide a full range of other plant analysis services to meet your different needs. If you are interested in us, please feel free to contact us.