A Microfluidic Chip-Based Method for Rapid Neutrophil Chemotaxis Analysis Using Whole Blood
Trascrizione
To a whole blood sample, add specific antibody complexes directed against blood cells except neutrophils.
Add magnetic particles that bind to the antibody complex-bound blood cells.
Transfer the immunomagnetically tagged sample to the cell-loading port of a microfluidic chip.
Attach magnetic disks to the cell-loading port.
The magnetic field attracts the immunomagnetically-tagged cells, trapping them to the port's side walls.
The neutrophils, being untagged, flow into the chip and become trapped at the cell-docking area.
Add fluorescently-labeled chemoattractant solution and migration medium to designated reservoirs. The liquids flow through the microfluidic channels, forming a concentration gradient.
When the chemoattractant binds to the neutrophil receptor, it triggers intracellular changes, causing neutrophils to reorganize their cytoskeleton and extend pseudopods.
These pseudopods act as feet, propelling the neutrophils toward higher chemoattractant concentrations — a process called chemotaxis.
Use appropriate microscopy techniques to monitor neutrophil migration in response to the chemoattractant gradient.
