Real-Time Deformability Cytometry

Whole blood sample flowing through an RT-DC chip

a) The RT-DC microfluidic chip is made out of PDMS assembled on an inverted microscope. b) Using a syringe pump a cell suspension is driven through a narrow constriction where cells are exposed to hydrodynamic shear stress and pressure. c) Deformation and cell sizes are recorded in real-time. d) Analytical and numerical models developed in our lab also permit the derivation of material properties such as the Young’s modulus [1, 2]. e) Mechanical fingerprint of a blood sample. Click image to enlarge.
Real-time deformability cytometry (RT-DC) is a novel high-throughput method for the mechanical characterization of single cells that has recently been developed in our lab [3]. Based on the hydrodynamic deformation of cells translocating through a microfluidic channel in a contact-free manner, RT-DC is able to analyze more than 100 cells per second in real-time.

The main working principle of RT-DC is shown in the figure. Within a few milliseconds upon entry of a cell into the channel, the cell shape reaches a steady state. For each cell, several parameters can be recorded in real-time which can be visualized and gated in a post-processing step using the custom-made software ShapeOut. Analytical and numerical models developed in our lab also permit the derivation of material properties such as the Young’s modulus [1, 2]. The technique is now being applied in more than 50 collaborations, e.g. to establish a mechanical fingerprint of blood.

Furthermore, we have recently developed an RT-DC setup with fluorescence detection (RT-FDC). Now it is possible to not only detect the mechanical phenotype of each individual cell but also to simultaneously gather its fluorescence intensity in up to three channels in a manner similar to a conventional flow cytometer. This direct correlation of mechanical with fluorescence data based on dyes, fluorescent reporter proteins or surface markers will lead to a more comprehensive validation of cell mechanics as a label-free marker.

RT-DC and RT-FDC are available as commercial products from the spin-off company ZELLMECHANIK DRESDEN GmbH.

  1. A. Mietke, O. Otto, S. Girardo, P. Rosendahl, A. Taubenberger, S. Golfier, E. Ulbricht, S. Aland, J. Guck, and E. Fischer-Friedrich, “Extracting cell stiffness from real-time deformability cytometry: theory and experiment,” Biophysical Journal, vol. 109, iss. 10, pp. 2023-2036, 2015. doi:10.1016/j.bpj.2015.09.006

    Title = {Extracting Cell Stiffness from Real-Time Deformability Cytometry: Theory and Experiment},
    Author = {Alexander Mietke and Oliver Otto and Salvatore Girardo and Philipp Rosendahl and Anna Taubenberger and Stefan Golfier and Elke Ulbricht and Sebastian Aland and Jochen Guck and Elisabeth Fischer-Friedrich},
    Journal = {{Biophysical Journal}},
    Year = {2015},
    Month = {nov},
    Number = {10},
    Pages = {2023--2036},
    Volume = {109},
    Doi = {10.1016/j.bpj.2015.09.006},
    Owner = {paul},
    Publisher = {Elsevier {BV}},
    Timestamp = {2016.09.17}

  2. M. Mokbel, D. Mokbel, A. Mietke, N. Träber, S. Girardo, O. Otto, J. Guck, and S. Aland, “Numerical simulation of real-time deformability cytometry to extract cell mechanical properties,” Acs biomaterials science & engineering, 2017. doi:10.1021/acsbiomaterials.6b00558
    [BibTeX] [Download PDF]

    Title = {Numerical Simulation of Real-Time Deformability Cytometry to Extract Cell Mechanical Properties},
    Author = {Mokbel, Marcel and Mokbel, Dominic and Mietke, Alexander and Tr{\"a}ber, Nicole and Girardo, Salvatore and Otto, Oliver and Guck, Jochen and Aland, Sebastian},
    Journal = {ACS Biomaterials Science \& Engineering},
    Year = {2017},
    Doi = {10.1021/acsbiomaterials.6b00558},
    Publisher = {{ACS Publications}},
    Url = {}

  3. O. Otto, P. Rosendahl, A. Mietke, S. Golfier, C. Herold, D. Klaue, S. Girardo, S. Pagliara, A. Ekpenyong, A. Jacobi, M. Wobus, N. Töpfner, U. F. Keyser, J. Mansfeld, E. Fischer-Friedrich, and J. Guck, “Real-time deformability cytometry: on-the-fly cell mechanical phenotyping,” Nature Methods, vol. 12, iss. 3, pp. 199-202, 2015. doi:10.1038/nmeth.3281

    Title = {Real-time deformability cytometry: on-the-fly cell mechanical phenotyping},
    Author = {Oliver Otto and Philipp Rosendahl and Alexander Mietke and Stefan Golfier and Christoph Herold and Daniel Klaue and Salvatore Girardo and Stefano Pagliara and Andrew Ekpenyong and Angela Jacobi and Manja Wobus and Nicole Töpfner and Ulrich F Keyser and Jörg Mansfeld and Elisabeth Fischer-Friedrich and Jochen Guck},
    Journal = {{Nature Methods}},
    Year = {2015},
    Month = {feb},
    Number = {3},
    Pages = {199--202},
    Volume = {12},
    Doi = {10.1038/nmeth.3281},
    Owner = {paul},
    Publisher = {Nature Publishing Group},
    Timestamp = {2016.09.17}