Real-time deformability cytometry as a label-free indicator of cell function

The mechanical properties of cells are known to be a label-free, inherent marker of biological function in health and disease. Wide-spread utilization has so far been impeded by the lack of a convenient measurement technique with sufficient throughput. To address this unmet need, we have recently introduced real-time deformability cytometry (RT-DC) for continuous mechanical single-cell classification of heterogeneous cell populations at rates of several hundred cells per second. Cells are driven through the constriction zone of a microfluidic chip leading to cell deformations due to hydrodynamic stresses only. Our custom-built image processing software performs image acquisition, image analysis and data storage on the fly. The ensuing deformations can be quantified and an analytical model enables the derivation of cell material properties. Performing RT-DC we highlight its potential to identify rare objects in heterogeneous suspensions and to track drug-induced changes in cells. In summary, RT-DC enables marker-free, quantitative phenotyping of heterogeneous cell populations with a throughput comparable to standard flow cytometry.

[1]  Carlota Saldanha,et al.  Multidisciplinary utilization of dimethyl sulfoxide: pharmacological, cellular, and molecular aspects. , 2003, Biochemical pharmacology.

[2]  Subra Suresh,et al.  The biomechanics toolbox: experimental approaches for living cells and biomolecules , 2003 .

[3]  O. Thoumine,et al.  Comparison of the mechanical properties of normal and transformed fibroblasts. , 1997, Biorheology.

[4]  Subra Suresh,et al.  Biomechanics and biophysics of cancer cells. , 2007, Acta biomaterialia.

[5]  E. Elson,et al.  Cellular mechanics as an indicator of cytoskeletal structure and function. , 1988, Annual review of biophysics and biophysical chemistry.

[6]  D. Stamenović,et al.  Modulation of Cellular Mechanics during Osteogenic Differentiation of Human , 2004 .

[7]  Stefan Schinkinger,et al.  Optical deformability as an inherent cell marker for testing malignant transformation and metastatic competence. , 2005, Biophysical journal.

[8]  Daniel A. Fletcher,et al.  Cell mechanics and the cytoskeleton , 2010, Nature.

[9]  Falk Wottawah,et al.  Oral cancer diagnosis by mechanical phenotyping. , 2009, Cancer research.

[10]  Dino Di Carlo,et al.  Hydrodynamic stretching of single cells for large population mechanical phenotyping , 2012, Proceedings of the National Academy of Sciences.

[11]  Stefan Schinkinger,et al.  The regulatory role of cell mechanics for migration of differentiating myeloid cells , 2009, Proceedings of the National Academy of Sciences.

[12]  Z. Stachura,et al.  Elasticity of normal and cancerous human bladder cells studied by scanning force microscopy , 1999, European Biophysics Journal.

[13]  Keiichi Abe,et al.  Topological structural analysis of digitized binary images by border following , 1985, Comput. Vis. Graph. Image Process..

[14]  W. Pfeffer,et al.  Osmotische Untersuchungen : studien zur zellmechanik , 1921 .

[15]  Andrew E. Pelling,et al.  An historical perspective on cell mechanics , 2008, Pflügers Archiv - European Journal of Physiology.

[16]  Jochen Guck,et al.  Viscoelastic Properties of Differentiating Blood Cells Are Fate- and Function-Dependent , 2012, PloS one.

[17]  Subra Suresh,et al.  Biomechanics and biophysics of cancer cells q , 2007 .

[18]  U. Keyser,et al.  Real-time deformability cytometry: on-the-fly cell mechanical phenotyping , 2015, Nature Methods.

[19]  Jaydev P Desai,et al.  Mechanical phenotyping of stem cells. , 2011, Theriogenology.

[20]  Yo Sup Moon,et al.  Quantitative Diagnosis of Malignant Pleural Effusions by Single-Cell Mechanophenotyping , 2013, Science Translational Medicine.

[21]  Sanjay Kumar,et al.  Mechanics, malignancy, and metastasis: The force journey of a tumor cell , 2009, Cancer and Metastasis Reviews.

[22]  Brenton D Hoffman,et al.  Cell mechanics: dissecting the physical responses of cells to force. , 2009, Annual review of biomedical engineering.

[23]  David A Weitz,et al.  The cell as a material. , 2007, Current opinion in cell biology.

[24]  Rajarshi Pal,et al.  Diverse effects of dimethyl sulfoxide (DMSO) on the differentiation potential of human embryonic stem cells , 2011, Archives of Toxicology.

[25]  Y. Nakajima,et al.  Involvement of mitochondrial permeability transition and caspase-9 activation in dimethyl sulfoxide-induced apoptosis of EL-4 lymphoma cells. , 2001, International immunopharmacology.

[26]  Oliver Otto,et al.  Extracting Cell Stiffness from Real-Time Deformability Cytometry: Theory and Experiment , 2015, Biophysical journal.