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Enhanced functionalities of immune cells separated by a microfluidic lattice: assessment based on holotomography
Mahn Jae Lee1,2, Byungyeon Kim3, Dohyeon Lee2,4
1Graduate School of Medical Science and Engineering, Korea Advanced Institute of Science and Technology (KAIST), Daejeon 34141, Republic of Korea.
Insights
Conventional centrifugation harms white blood cell (WBC) function, while microfluidic separation preserves it. This study reveals microfluidics as a superior method for isolating functional WBCs for research and diagnostics.
Area of Science:
- Immunology
- Cell Biology
- Biomedical Engineering
Background:
- White blood cell (WBC) isolation is crucial for immunology, diagnostics, and immunotherapy.
- The impact of traditional density gradient centrifugation on WBC functionality is not well understood.
Purpose of the Study:
- To compare the effects of conventional centrifugation and microfluidic separation on isolated WBC functionality.
- To assess morphology, motility, and phagocytic capabilities using holotomography.
Main Methods:
- Holotomography was used to analyze individual WBCs.
- Three-dimensional refractive index distribution and time-lapse dynamics were employed.
- WBCs were isolated using conventional centrifugation and microfluidic separation.
Main Results:
- Conventional centrifugation negatively affected WBC motility and phagocytic capacity.
- Microfluidic separation demonstrated a more favorable outcome in preserving WBC functionality.
- Holotomography provided in-depth analysis of individual cell behavior.
Conclusions:
- Microfluidic separation is a promising alternative to centrifugation for preserving WBC functionality.
- This technique can enhance precision in immunological studies and diagnostic accuracy for hematologic disorders.
Abstract:
The isolation of white blood cells (WBCs) from whole blood constitutes a pivotal process for immunological studies, diagnosis of hematologic disorders, and the facilitation of immunotherapy. Despite the ubiquity of density gradient centrifugation in WBC isolation, its influence on WBC functionality remains inadequately understood. This research employs holotomography to explore the effects of two distinct WBC separation techniques, namely conventional centrifugation and microfluidic separation, on the functionality of the isolated cells. We utilize three-dimensional refractive index distribution and time-lapse dynamics to analyze individual WBCs in-depth, focusing on their morphology, motility, and phagocytic capabilities. Our observations highlight that centrifugal processes negatively impact WBC motility and phagocytic capacity, whereas microfluidic separation yields a more favorable outcome in preserving WBC functionality. These findings emphasize the potential of microfluidic separation techniques as a viable alternative to traditional centrifugation for WBC isolation, potentially enabling more precise analyses in immunology research and improving the accuracy of hematologic disorder diagnoses.
