Real time unlabeled cell analyzer

xCELLigence RTCA S16Real time unlabeled cell analyzer (1x16)Real time continuous monitoring of live cell proliferation, morphological changes, and adhesion strength through label free methods using biosensors.

xCELLigence RTCA S16Real time unlabeled cell analyzerOperate in a standard CO2 cell culture incubator, with the workstation located outside the incubator. User friendly software can achieve real-time control and monitoring of instruments, and has real-time data analysis functions.

characteristic

1. Obtain continuous cell analysis time points throughout the entire experimental process.

2. RTCA can provide continuous quantitative readings of cell quantity, proliferation rate, cell size/shape, and cell matrix adhesion strength.

technology

Cell impedance analysis

Cell detection method is an important tool in basic and applied biological research, which lies between reductionist biochemical detection and whole organism in vivo experiments. However, the practicality of many cell detection methods is reduced due to the following factors: (1) the need for markers, (2) the inability to perform continuous monitoring (i.e. only generate endpoint data), (3) incompatibility with other orthogonal detection methods, and (4) the inability to provide objective/quantitative readings. However, each of these drawbacks is overcome by non-invasive, label free, and real-time cell impedance detection methods.

Functional unit for cell impedance detection

The functional unit of cell impedance detection is a set of gold microelectrodes fused to the bottom surface of the microtiter plate well (Figure 1). When these electrodes are immersed in a conductive solution (such as buffer or standard tissue culture medium), applying a potential across the electrodes causes electrons to flow out of the negative electrode, pass through the solution itself, and then deposit on the positive electrode, thus completing the circuit. Due to the dependence of this phenomenon on the interaction between the electrode and the solution itself, the presence of adherent cells at the electrode solution interface can hinder electron flow. The magnitude of this impedance depends on the number of cells, the size and shape of the cells, and the quality of their attachment to the matrix. Importantly, the surface of the gold micro electrode or the applied potential (22 millivolts) will not affect the health or behavior of the cells.

Impedance electrode

In each well of Agient's E-plates, the gold microelectrode biosensor covers 70% -80% of the surface area (depending on the presence of the observation area). Unlike the simplified electrode pair shown in Figure 1, the electrodes in each hole of E-Plates are connected in a "strip" to form a staggered array (Figure 2). This arrangement allows for simultaneous monitoring of cell populations, enabling extremely sensitive detection of the following indicators: the number of cells attached to the plate, the size/morphology of cells, and the quality of cell attachment to the matrix.

Real time impedance curve analysis

The electron flow impedance caused by cell adhesion is represented by a dimensionless parameter, the Cell Index (CI), where CI=(impedance at time point n - impedance without cells)/nominal impedance value. Figure 3 provides a general example of real-time impedance curves during the setup and operation of apoptosis experiments.

In the first few hours after cells are added to the culture well, impedance rapidly increases. This is because cells fall from a suspended state, deposit on the electrode, and form focal adhesion. If the initial number of cells added is small and there is free space at the bottom of the culture well, the cells will proliferate, leading to a gradual and stable increase in cell index (CI). When the cells reach a confluent state, the CI value tends to stabilize, reflecting that the portion of the electrode surface area that can be contacted by a large amount of culture medium no longer changes. Adding apoptosis inducers at this time will cause the CI value to decrease to zero. This is because the cells become round and detach from the bottom of the culture well. Although this general example involves adding drugs during cell confluence, impedance based measurement methods are very flexible and can also evaluate the rate and degree of initial cell adhesion to electrodes, or the rate and degree of cell proliferation.

The correlation between impedance and cellular phenomena

Real time cell analysis (RTCA) technology provides a quantitative means of detecting cell quantity, proliferation rate, cell size/morphology, and cell matrix adhesion quality. Due to the fact that these physical properties are a comprehensive reflection of thousands of different gene/protein products, RTCA technology can widely reflect the health status and behavior of cells. From endothelial barrier function and chemotaxis to filamentous pseudopodia dynamics and immune cell-mediated cell lysis, the xCELLigence instrument has successfully analyzed numerous cellular phenomena. Despite its wide range of applications, xCELLigence detection can still explore very specific biochemical and cellular phenomena. By using appropriate control and/or orthogonal techniques, we can associate the characteristics of impedance curves with specific cellular/molecular phenomena. To gain a deeper understanding of how this association is established and witness the sensitivity and versatility of xCELLigence RTCA technology, please browse through the many specific applications highlighted here.

working principle

Real time detection of cell behavior under physiological conditions

The xCELLigence RTCA instrument uses impedance based technology to continuously and automatically detect the health and behavior of cells in a standard CO2 cell culture incubator within seconds to days. Simply add the cells to the E-Plate to start real-time monitoring of cell count, cell proliferation rate, cell size, cell morphology, and cell matrix adhesion strength.

Explanation of Cell Impedance Technology

This high-sensitivity real-time cell impedance analysis uses a micro gold electrode fused to the bottom of the E-Plate, which can non invasively detect cell characteristics. The presence of adherent cells at the interface between electrodes and solution can hinder electron flow. The strength of impedance depends on the number, size, shape, and adhesion strength between cells and matrix. The surface of the micro gold electrode and the applied potential (22 mV) will not affect cell viability and behavior.

Simple and stable detection workflow

The xCELLigence RTCA workflow can help users obtain highly sensitive and reproducible quantitative analysis data, while greatly reducing manual processing time. Just add the cells to the E-Plate well plate to start real-time detection of cell quantity, cell proliferation rate, cell size, cell morphology, and cell matrix adhesion strength. Use the Cell Index, a non unit parameter, to report the cell current impedance caused by adherent cells.

application

Immune cell killing detection

Conventional immune cell killing detection is time-consuming and labor-intensive, requiring users to use multiple technology combinations to collect data at multiple time points. In contrast, the unlabeled xCELLigence RTCA instrument provides comprehensive information on immune cell killing and approximates in vivo activity. Real time detection of immune cell killing efficacy with high sensitivity and reproducibility. This simple high-throughput workflow allows for in-depth study of cellular mechanisms, continuous killing, and exhaustion.

Virus lesion effect detection

Use Agilent xCELLigence RTCA instrument to accurately detect the cytopathic effects (CPE) of viruses in real-time, advancing your antiviral therapy or vaccine development research without the need for time-consuming and laborious plaque experiments. Automatically monitor virus CPE within minutes to days, significantly reducing workload and manual sample processing. Detect cell proliferation kinetics, determine the optimal time point for viral infection at different cell inoculation densities, and evaluate virus mediated cellular lesions in the workflow.

Cytotoxicity analysis

Determine the cytotoxic effects of drugs or compounds by real-time monitoring of cell proliferation, cell size or morphology, and cell matrix adhesion strength. Use this simple and high-throughput cytotoxicity analysis to monitor the kinetics of long-term cellular responses. Make wise decisions on cell processing time without increasing workload, and easily generate dose-response curves at multiple time points.

Cell adhesion detection

Primary cancer involves millions of cells shedding into the circulatory system during tumor formation, known as circulating tumor cells (CTCs). CTCs can reattach, spread, and metastasize to distant organs. By using xCELLigence RTCA, we can continuously monitor the adhesion kinetics of different types of cells and gain a deeper understanding of the complexity of metastasis. Real time quantitative evaluation of cell adhesion and extension, without the need for fluorescent reagents or dyes.

Cell signal transduction detection

G protein coupled receptor (GPCR), receptortyrosine kinaseRTK or nuclear hormone mediated signal transduction often leads to biochemical changes, affecting the number of cells present, cell size/morphology, and the degree of interaction between cells and the surface of the well plate. By using xCELLigence RTCA, you can sensitively capture changes in cell size, morphology, and proliferation rate with cellular signaling events. This label free continuous monitoring can quantitatively evaluate cellular responses in the first few minutes and days after treatment.

Stem cell testing

Stem cell research has helped people understand disease mechanisms, contributed to the development of regenerative medicine, and provided an alternative method for testing the reliability and effectiveness of new drugs. Using xCELLigence RTCA to predict the functionality of stem cell products and evaluate the variability of mesenchymal stem cells (MSCs) to overcome challenges in the production of cell therapy products. Significantly improving analysis efficiency with fewer cells, continuously detecting comprehensive changes in cell quantity, adhesion strength, and morphology to ensure consistency at each stage.

Analysis of cellular barrier function

The cell barrier function is provided by endothelial cells and epithelial cells, and can be impaired in many disease states. Endothelial cells are arranged on the inner surface of blood vessels and lymphatic vessels. Epithelial cells form the surface of our body, serving as a barrier to protect the body from damage from foreign molecules. The impedance based xCELLigence RTCA detection method is a sensitive quantitative approach that can replace traditional methods such as solute permeability detection and transendothelial cell resistance (TEER) detection to detect barrier disruption.