Centrifugal microfluidics for sorting immune cells from whole blood

Abstract

Sorting and enumeration of immune cells from blood are critical operations involved in many clinical applications. Conventional methods for sorting and counting immune cells from blood, such as flow cytometry and hemocytometers, are tedious, inaccurate, and difficult for implementation for point-of-care (POC) testing. Herein we developed a microscale centrifugal technology termed Centrifugal Microfluidic Chip (CMC) capable of sorting immune cells from blood and in situ cellular analysis in a laboratory setting. Operation of the CMC entailed a blood specimen layered on a density gradient medium and centrifuged in microfluidic channels where immune cell subpopulations could rapidly be sorted into distinct layers according to their density differentials. We systematically studied effects of different blocking molecules for surface passivation of the CMC. We further demonstrated the applicability of CMCs for rapid separation of minimally processed human whole blood without affecting immune cell viability. Multi-color imaging and analysis of immune cell distributions and enrichment such as recovery and purity rates of peripheral blood mononuclear cells (PBMCs) were demonstrated using CMCs. Given its design and operation simplicity, portability, blood cell sorting efficiency, and in situ cellular analysis capability, the CMC holds promise for blood-based diagnosis and disease monitoring in POC applications.

A Review of Design Considerations for Hemocompatibility within Microfluidic Systems

“…A comparison between PEG, PLL-g-PEG, and trypsin (an uncommon surface passivator) in a microfluidic centrifugal cell sorter demonstrated similar efficacies of PLL-PEG and Trypsin in preventing cellular adhesion, while PEG was not as effective. 90 Self-Assembled Monolayers Self-assembled monolayers (SAMs), illustrated in ►Fig. 2(A5) are ordered molecular assemblies produced by a spontaneous binding of active chemical moieties to reactive substrates.…”

The Role of Micro fluidic Systems in Biological and Medical Sciences

“…Micro fluidic chips have been used in numerous applications, for example as platform for infrared absorption [13], cell culture [14,15], droplet formation [16], digital PCR [17], electrophoresis [18,19] and biosensors [20]. Yu et al [21] employed chips to separate and sort cells by micro centrifugation. Joshi et al [22] used them to fabricate liposomes as carrier for pharmaceutical drugs.…”

Design and Simulation of an Integrated Centrifugal Microfluidic Device for CTCs Separation and Cell Lysis

“…Microfluidics is a proven technology that has been employed to create niche solutions to biomedical applications, such as cell separation and mixing, 3D bioprinting [ 7 , 8 , 9 , 10 ], and organs-on-a-chip systems [ 11 , 12 ]. The application of microfluidic technologies can address some of the limitations of mentioned commercial cell separation methods by using different physical mechanisms, including filtration- [ 13 ], hydrodynamic- [ 14 ], inertial- [ 15 ], deterministic lateral displacement (DLD)- [ 16 , 17 ], pinched flow fractionation (PFF)- [ 18 ], centrifugation- [ 19 ], dielectrophoresis (DEP)- [ 20 ], magnetic- [ 21 ], acoustic- [ 22 ], and optical-based approaches [ 23 ]. These methods can separate target cells from a heterogeneous cell population by exploiting the differences in the properties of the cells, including their size, density, shape, deformability, and compressibility, as well as their electric, magnetic, and optical properties.…”

“…Centrifugal microfluidics have been utilized in numerous studies, in which the separation of blood components are needed. This includes plasma separation from blood cells [ 30 ] and the extraction of leukocytes from blood samples [ 31 ] and, additionally, to separate immune cells [ 19 ] and isolate CTCs from whole blood [ 32 ].…”