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Research on the Method of Detecting TPN-Labeled Tumor Cells in Pleural Effusion Based on the Microfluidic Chip
Xiaoyi Xun1, Shuang Song2, Yiran Luan1
1School of Medical Laboratory, Tianjin Medical University, Tianjin 300203, China.
Abstract:
The clinical diagnosis of a malignant pleural effusion (MPE) is still based on the detection of tumor cells in the pleural effusion. The question of how to improve the efficiency and accuracy of detecting an MPE still remains. This study explores the use of microfluidic technology to concentrate cells in an MPE and achieved the detection of the cell marker TPN in the microarray capture area. TPN is a mitochondria-specific bio-probe that can identify tumor cells on the basis of differences in the mitochondrial potential. First, we designed a microfluidic chip to analyze its performance. The results show that when the total flow rate of the injected chip was 12 mL/h and the volume ratio of cell separation liquid to cell suspension was 1:1, the target cells (A549, MCF-7, and Hela) were enriched and the purity was improved to 98.7-99.3%. Finally, an MPE from cancer patients was used to detect the chip's ability to isolate and enrich tumor cells. Furthermore, the fluorescent identification of the TPN within the tumor cells was simultaneously achieved on the microfluidic chip. In conclusion, the potential to improve the efficiency of the clinical diagnosis of MPEs is provided by the chip structure and analysis conditions explored in this study.
Insights
This study introduces a microfluidic chip for improved malignant pleural effusion (MPE) diagnosis. The technology efficiently concentrates tumor cells and detects the TPN marker, enhancing diagnostic accuracy.
Area of Science:
- Biomedical Engineering
- Oncology
- Cell Biology
Background:
- Malignant pleural effusion (MPE) diagnosis relies on detecting tumor cells in pleural fluid.
- Current diagnostic methods for MPE require improvement in efficiency and accuracy.
Purpose of the Study:
- To develop and evaluate a microfluidic chip for concentrating tumor cells from MPE.
- To enable simultaneous detection of the TPN cell marker for improved MPE diagnosis.
Main Methods:
- Design and performance analysis of a novel microfluidic chip.
- Optimization of flow rate and liquid ratios for cell enrichment.
- Utilizing a mitochondria-specific bio-probe (TPN) for tumor cell identification.
- Testing the chip with MPE samples from cancer patients.
Main Results:
- The microfluidic chip achieved high purity (98.7-99.3%) in enriching target cells (A549, MCF-7, Hela).
- Optimal conditions identified: 12 mL/h total flow rate and 1:1 volume ratio of cell separation liquid to cell suspension.
- Successful isolation and enrichment of tumor cells from clinical MPE samples.
- Simultaneous fluorescent identification of TPN within tumor cells on the chip.
Conclusions:
- The developed microfluidic chip shows significant potential for improving MPE diagnostic efficiency.
- The optimized chip structure and analysis conditions enhance tumor cell detection in MPE.
- This technology offers a promising advancement for the clinical diagnosis of malignant pleural effusions.
