Measuring single-cell density

Grover, William H.; Bryan, Andrea K.; Diez-Silva, Monica; Suresh, S.; Higgins, John M.; Manalis, Scott R.

doi:10.1073/pnas.1104651108

Cited by 297 publications

(260 citation statements)

References 19 publications

Supporting

Mentioning

252

Contrasting

Order By: Relevance

“…Based on the density of 1.08 g/cm 3 reported for lymphocytes 38 and the density (1.02 g/cm 3 ) of the DEP medium, v s can be estimated as $3 lm/s. Even in the absence of a negative DEP force driving cells downward from the upper plane of electrodes, there was thus a high probability that cells passing through the chamber would sediment down to the lower electrode plane and thereby not escape strong DEP forces.…”

Section: Discussionmentioning

confidence: 99%

Biomarker-free dielectrophoretic sorting of differentiating myoblast multipotent progenitor cells and their membrane analysis by Raman spectroscopy

et al. 2012

View full text Add to dashboard Cite

Myoblasts are muscle derived mesenchymal stem cell progenitors that have great potential for use in regenerative medicine, especially for cardiomyogenesis grafts and intracardiac cell transplantation. To utilise such cells for pre-clinical and clinical applications, and especially for personalized medicine, it is essential to generate a synchronised, homogenous, population of cells that display phenotypic and genotypic homogeneity within a population of cells. We demonstrate that the biomarker-free technique of dielectrophoresis (DEP) can be used to discriminate cells between stages of differentiation in the C2C12 myoblast multipotent mouse model. Terminally differentiated myotubes were separated from C2C12 myoblasts to better than 96% purity, a result validated by flow cytometry and Western blotting. To determine the extent to which cell membrane capacitance, rather than cell size, determined the DEP response of a cell, C2C12 myoblasts were co-cultured with GFP-expressing MRC-5 fibroblasts of comparable size distributions (mean diameter $10 lm). A DEP sorting efficiency greater than 98% was achieved for these two cell types, a result concluded to arise from the fibroblasts possessing a larger membrane capacitance than the myoblasts. It is currently assumed that differences in membrane capacitance primarily reflect differences in the extent of folding or surface features of the membrane. However, our finding by Raman spectroscopy that the fibroblast membranes contained a smaller proportion of saturated lipids than those of the myoblasts suggests that the membrane chemistry should also be taken into account.

show abstract

Section: Discussionmentioning

confidence: 99%

Biomarker-free dielectrophoretic sorting of differentiating myoblast multipotent progenitor cells and their membrane analysis by Raman spectroscopy

et al. 2012

View full text Add to dashboard Cite

show abstract

“…In Hb CC, the entire distribution of densities of erythrocytes shifts to a slightly higher density; reticulocytes in Hb CC are denser than those in Hb AA, but the high-density erythrocytes in Hb CC may be less dense than the densest cells in Hb SS (12). In SCD, erythrocytes that are not sickled remain at normal densities, and the erythrocytes exist in two populations: a high-density, often sickled, population (∼10% of cells) and a lower-density, predominantly normocytic, population, which also comprises reticulocytes and the youngest erythrocytes (24,25).…”

Section: Scd Refers To a Large Number Of Genetic Variationsmentioning

confidence: 99%

Density-based separation in multiphase systems provides a simple method to identify sickle cell disease

Kumar¹,

Patton²,

Hennek³

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

106

120

View full text Add to dashboard Cite

Although effective low-cost interventions exist, child mortality attributable to sickle cell disease (SCD) remains high in low-resource areas due, in large part, to the lack of accessible diagnostic methods. The presence of dense (ρ > 1.120 g/cm 3 ) cells is characteristic of SCD. The fluid, self-assembling step-gradients in density created by aqueous multiphase systems (AMPSs) identifies SCD by detecting dense cells. AMPSs separate different forms of red blood cells by density in a microhematocrit centrifuge and provide a visual means to distinguish individuals with SCD from those with normal hemoglobin or with nondisease, sickle-cell trait in under 12 min. Visual evaluation of a simple two-phase system identified the two main subclasses of SCD [homozygous (Hb SS) and heterozygous (Hb SC)] with a sensitivity of 90% (73-98%) and a specificity of 97% (86-100%). A threephase system identified these two types of SCD with a sensitivity of 91% (78-98%) and a specificity of 88% (74-98%). This system could also distinguish between Hb SS and Hb SC. To the authors' knowledge, this test demonstrates the first separation of cells by density with AMPSs, and the usefulness of AMPSs in point-of-care diagnostic hematology.polymers | erythrocytes | sickle cell anemia | cell sorting | density gradient centrifugation

show abstract

“…1 However, the measurement of single cellular mass and density has proven to be very difficult for biological researchers. The main reason is that the mass of a micron-size cell is very small-about hundreds of picograms-which makes the determination of cell mass very difficult, especially in a fluidic environment.…”

Section: Introductionmentioning

confidence: 99%

Measurement of single leukemia cell's density and mass using optically induced electric field in a microfluidics chip

et al. 2015

View full text Add to dashboard Cite

We present a method capable of rapidly ($20 s) determining the density and mass of a single leukemic cell using an optically induced electrokinetics (OEK) platform. Our team had reported recently on a technique that combines sedimentation theory, computer vision, and micro particle manipulation techniques on an OEK microfluidic platform to determine the mass and density of micron-scale entities in a fluidic medium; the mass and density of yeast cells were accurately determined in that prior work. In the work reported in this paper, we further refined the technique by performing significantly more experiments to determine a universal correction factor to Stokes' equation in expressing the drag force on a microparticle as it falls towards an infinite plane. Specifically, a theoretical model for micron-sized spheres settling towards an infinite plane in a microfluidic environment is presented, and which was validated experimentally using five different sizes of micro polystyrene beads. The same sedimentation process was applied to two kinds of leukemic cancer cells with similar sizes in an OEK platform, and their density and mass were determined accordingly. Our tests on mouse lymphocytic leukemia cells (L1210) and human leukemic cells (HL-60) have verified the practical viability of this method. Potentially, this new method provides a new way of measuring the volume, density, and mass of a single cell in an accurate, selective, and repeatable manner. V C 2015 AIP Publishing LLC. [http://dx

show abstract

Measuring single-cell density

Cited by 297 publications

References 19 publications

Biomarker-free dielectrophoretic sorting of differentiating myoblast multipotent progenitor cells and their membrane analysis by Raman spectroscopy

Biomarker-free dielectrophoretic sorting of differentiating myoblast multipotent progenitor cells and their membrane analysis by Raman spectroscopy

Density-based separation in multiphase systems provides a simple method to identify sickle cell disease

Measurement of single leukemia cell's density and mass using optically induced electric field in a microfluidics chip

Contact Info

Product

Resources

About