On-Chip Continuous Blood Cell Subtype Separation by Deterministic Lateral Displacement

Abstract

This paper presents a microfluidic device for continuous human blood cell subtype separation using the deterministic lateral displacement principle. Based on their significant size and shape differences, three major cell types of human whole blood -platelets, red blood cells and white blood cells -were demonstrated to be directly separated using a twostage separation strategy. Even though all white blood cells are spherical and have diameters within a narrow range (8 -20pm), the initial limitation for using this principle to separate white blood cell subtypes was conquered by attaching larger polystyrene microbeads to one of the subtypes to amplify the size differences. Specifically, continuous separation of human CD4+ T helper lymphocytes from other white blood cell subtypes was achieved with high purity and recovery due to the underlying high affinity and high specificity of the antigen-antibody interaction used to attach the microbeads to the lymphocytes.With our novel approach, the pure population of one blood cell subtype can be effectively isolated by exploring the deterministic lateral displacement principle, which has the advantages of the simplicity, high speed and high resolution. Because many cells express unique surface markers, this method can theoretically be applied to separate any target cell type from a heterogeneous mixture for downstream analysis.

Centrifuge-based deterministic lateral displacement separation

“…More recently, it has been shown that DLD can also fractionate samples based on shape and deformability (Quek et al 2011;Bogunovic et al 2012;Beech et al 2012;Zeming et al 2013;Ranjan et al 2014;Krüger et al 2014;Holmes et al 2014). DLD has been successful in the fractionation of various mixtures, including biological samples and cells in particular (Beech et al 2012;Zeming et al 2013;Ranjan et al 2014;Krüger et al 2014;Holmes et al 2014;Siyang Zheng et al 2005;Davis et al 2006;Nan Li et al 2007;Huang et al 2008;Inglis et al 2008;Morton et al 2008;Green et al 2009;Inglis et al 2010;Holm et al 2011;Al-Fandi et al 2011;Inglis et al 2011;Zhang et al 2012;Loutherback et al 2012;Liu et al 2013;Ozkumur et al 2013;Karabacak et al 2014), which makes it attractive for lab-on-a-CD applications. In addition, different post geometries (Loutherback et al 2010;Al-Fandi et al 2011;Ranjan et al 2014) and other design considerations (Inglis 2009) have been proposed to improve performance.…”

Cell separation by non-inertial force fields in microfluidic systems

“…This principle was applied to separate RBCs, WBCs, and platelets (Davis et al, 2006; Zheng et al, 2005a; Zheng et al, 2005b). More recently a subpopulation (CD4 + T helper lymphocytes) of WBCs was successfully fractionated by increasing the hydrodynamic radius through affinity binding of antibody-coated microbeads (Li et al, 2007). …”

Microtechnology meets systems biology: The small molecules of metabolome as next big targets

“…Strong, nonuniform and time varying electric fields can be generated on-chip without the need for applying high external potentials. It has been demonstrated in various applications, that DEP is suitable to separate different types of mammalian cells (Yang et al, 2000(Yang et al, , 1999bGiddings, 1993;Rousselet et al, 1998;Kersaudy-Kerhoas et al, 2008;Huang et al, 1999), yeast (Doh et al, 2004;Li et al, 2007), bacteria (Lapizco-Encinas et al, 2004;Hu et al, 2005) or polymeric particles (Rousselet et al, 1998;Kralj et al, 2006) into different fluid streams, which allows continuous or batch fractionation. DEP has been also successfully used for cell guiding and trapping (Muller et al, 1999;Voldman et al, 2002), which could be used for medium removal and cell washing.…”