Real-Time Deformability Cytometry


Whole blood sample flowing through an RT-DC chip

RTDC
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, 4, 5]. 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 our in-house 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 [6, 7, 8].

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 [9]. 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] [doi] 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, p. 2023–2036, 2015.
[Bibtex]
@Article{Mietke,
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] [doi] 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.
[Bibtex]
@Article{Mokbel2017,
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 = {http://pubs.acs.org/doi/pdfplus/10.1021/acsbiomaterials.6b00558}
}
[3] [doi] 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, p. 199–202, 2015.
[Bibtex]
@Article{Otto,
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}
}
[4] [doi] M. Herbig, M. Kräter, K. Plak, P. Müller, J. Guck, and O. Otto, “Real-time deformability cytometry: label-free functional characterization of cells,” in Flow cytometry protocols, Springer New York, 2017, p. 347–369.
[Bibtex]
@InCollection{Herbig_2017,
author = {Maik Herbig and Martin Kräter and Katarzyna Plak and Paul Müller and Jochen Guck and Oliver Otto},
title = {Real-Time Deformability Cytometry: Label-Free Functional Characterization of Cells},
booktitle = {Flow Cytometry Protocols},
publisher = {Springer New York},
year = {2017},
pages = {347--369},
month = {oct},
doi = {10.1007/978-1-4939-7346-0_15},
url = {https://doi.org/10.1007%2F978-1-4939-7346-0_15},
}
[5] [doi] M. Urbanska, P. Rosendahl, M. Kräter, and J. Guck, “High-throughput single-cell mechanical phenotyping with real-time deformability cytometry,” in Methods in cell biology, Elsevier, 2018, p. 175–198.
[Bibtex]
@InCollection{Urbanska_2018,
author = {Marta Urbanska and Philipp Rosendahl and Martin Kräter and Jochen Guck},
title = {High-throughput single-cell mechanical phenotyping with real-time deformability cytometry},
booktitle = {Methods in Cell Biology},
publisher = {Elsevier},
year = {2018},
pages = {175--198},
doi = {10.1016/bs.mcb.2018.06.009},
url = {https://doi.org/10.1016%2Fbs.mcb.2018.06.009},
}
[6] [doi] N. Toepfner, C. Herold, O. Otto, P. Rosendahl, A. Jacobi, M. Kräter, J. Stächele, L. Menschner, M. Herbig, L. Ciuffreda, L. Ranford-Cartwright, M. Grzybek, Ü. Coskun, E. Reithuber, G. Garriss, P. Mellroth, B. Henriques-Normark, N. Tregay, M. Suttorp, M. Bornhäuser, E. R. Chilvers, R. Berner, and J. Guck, “Detection of human disease conditions by single-cell morpho-rheological phenotyping of blood,” eLife, vol. 7, 2018.
[Bibtex]
@Article{Toepfner_2018,
author = {Nicole Toepfner and Christoph Herold and Oliver Otto and Philipp Rosendahl and Angela Jacobi and Martin Kräter and Julia Stächele and Leonhard Menschner and Maik Herbig and Laura Ciuffreda and Lisa Ranford-Cartwright and Michal Grzybek and Ünal Coskun and Elisabeth Reithuber and Genevi{\`{e}}ve Garriss and Peter Mellroth and Birgitta Henriques-Normark and Nicola Tregay and Meinolf Suttorp and Martin Bornhäuser and Edwin R Chilvers and Reinhard Berner and Jochen Guck},
title = {Detection of human disease conditions by single-cell morpho-rheological phenotyping of blood},
journal = {{eLife}},
year = {2018},
volume = {7},
month = {jan},
doi = {10.7554/elife.29213},
publisher = {{eLife} Sciences Publications, Ltd},
url = {https://elifesciences.org/download/aHR0cHM6Ly9jZG4uZWxpZmVzY2llbmNlcy5vcmcvYXJ0aWNsZXMvMjkyMTMvZWxpZmUtMjkyMTMtdjIucGRm/elife-29213-v2.pdf?_hash=kfISS47M%2B9T6A%2BkCMXxnzSQQUp%2B%2FILnUZ4qkUhMJKXE%3D},
}
[7] [doi] M. Kräter, J. Sapudom, N. Bilz, T. Pompe, J. Guck, and C. Claus, “Alterations in cell mechanics by actin cytoskeletal changes correlate with strain-specific rubella virus phenotypes for cell migration and induction of apoptosis,” Cells, vol. 7, iss. 9, p. 136, 2018.
[Bibtex]
@Article{Kr_ter_2018,
author = {Martin Kräter and Jiranuwat Sapudom and Nicole Bilz and Tilo Pompe and Jochen Guck and Claudia Claus},
title = {Alterations in Cell Mechanics by Actin Cytoskeletal Changes Correlate with Strain-Specific Rubella Virus Phenotypes for Cell Migration and Induction of Apoptosis},
journal = {Cells},
year = {2018},
volume = {7},
number = {9},
pages = {136},
month = {sep},
doi = {10.3390/cells7090136},
publisher = {{MDPI} {AG}},
url = {https://doi.org/10.3390%2Fcells7090136},
}
[8] [doi] M. Urbanska, M. Winzi, K. Neumann, S. Abuhattum, P. Rosendahl, P. Müller, A. Taubenberger, K. Anastassiadis, and J. Guck, “Single-cell mechanical phenotype is an intrinsic marker of reprogramming and differentiation along the mouse neural lineage,” Development, vol. 144, iss. 23, p. 4313–4321, 2017.
[Bibtex]
@Article{Urbanska_2017,
author = {Marta Urbanska and Maria Winzi and Katrin Neumann and Shada Abuhattum and Philipp Rosendahl and Paul Müller and Anna Taubenberger and Konstantinos Anastassiadis and Jochen Guck},
title = {Single-cell mechanical phenotype is an intrinsic marker of reprogramming and differentiation along the mouse neural lineage},
journal = {Development},
year = {2017},
volume = {144},
number = {23},
pages = {4313--4321},
month = {nov},
doi = {10.1242/dev.155218},
publisher = {The Company of Biologists},
url = {https://doi.org/10.1242%2Fdev.155218},
}
[9] [doi] P. Rosendahl, K. Plak, A. Jacobi, M. Kraeter, N. Toepfner, O. Otto, C. Herold, M. Winzi, M. Herbig, Y. Ge, S. Girardo, K. Wagner, B. Baum, and J. Guck, “Real-time fluorescence and deformability cytometry,” Nature methods, vol. 15, iss. 5, p. 355–358, 2018.
[Bibtex]
@Article{Rosendahl_2018,
author = {Philipp Rosendahl and Katarzyna Plak and Angela Jacobi and Martin Kraeter and Nicole Toepfner and Oliver Otto and Christoph Herold and Maria Winzi and Maik Herbig and Yan Ge and Salvatore Girardo and Katrin Wagner and Buzz Baum and Jochen Guck},
title = {Real-time fluorescence and deformability cytometry},
journal = {Nature Methods},
year = {2018},
volume = {15},
number = {5},
pages = {355--358},
month = {apr},
doi = {10.1038/nmeth.4639},
publisher = {Springer Nature},
url = {https://www.biorxiv.org/content/biorxiv/early/2017/09/11/187435.full-text.pdf},
}