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Published on: October 15, 2013
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Single Cell Analysis of Leukocyte Protease Activity Using Integrated Continuous-Flow Microfluidics
Tengyang Jing1,2, Zhangxing Lai1, Lidan Wu
1Department of Biomedical Engineering, National University of Singapore , Singapore 119077.
Analytical Chemistry
|November 1, 2016
Summary
A new microfluidic assay measures protease activity in individual leukocytes, revealing how stimulation affects immune cell function and heterogeneity. This technology aids in understanding immune responses and individual patient health.
Area of Science:
- Immunology
- Biochemistry
- Microfluidics
Background:
- Leukocytes are crucial immune cells defending against pathogens.
- Secretory products like matrix metalloproteinases (MMPs) and ADAMs regulate inflammation.
- Current methods cannot functionally assay protease activity in single, viable leukocytes.
Purpose of the Study:
- To develop a microfluidic assay for detecting individual leukocyte protease activity.
- To analyze single leukocyte protease profiles under different conditions.
Main Methods:
- Integrated continuous-flow microfluidic assay.
- On-chip washing of leukocytes to remove background activity.
- Encapsulation of single leukocytes with protease sensors in droplets for incubation and measurement.
Main Results:
- Reliable measurement of single leukocyte protease profiles.
- PMA stimulation increases average protease activity.
- PMA treatment reduces heterogeneity in leukocyte protease secretion.
Conclusions:
- The microfluidic assay effectively measures individual leukocyte protease secretion.
- Understanding protease activity and heterogeneity is vital for immune capability assessment.
- This technology can inform individual patient disease conditions and immune status.
