Abstract
In this paper we demonstrate how the use of frequencies ranging from 50 kHz to 5 GHz in the analysis of cells by electrorotation can open the path to the identification of differences not detectable by conventional set-ups. Earlier works usually reported electrorotation devices operating below 20 MHz, limiting the response obtained to properties associated with the cell membrane. Those devices are thus unable to resolve the physiological properties in the cytoplasm. We used microwave-based technology to extend the frequency operation to 5 GHz. At high frequencies (from tens of MHz to GHz), the electromagnetic signal passes through the membrane and allows probing the cytoplasm. This enables several applications, such as cell classification, and viability analysis. Additionally, the use of conventional microfabrication techniques reduces the cost and complexity of analysis, compared to other non-invasive methods. We demonstrated the potential of this set-up by identifying two different populations of T-lymphocytes not distinguishable through visual assessment. We also assessed the effect of calcein on cell cytoplasmic properties and used it as a controlled experiment to demonstrate the possibility of this method to detect changes happening predominantly in the cytoplasm.
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Abbreviations
- \(<>\) :
-
Time averaged
- A:
-
Radius of the circle circumscribed by the electrodes
- AC:
-
Alternating current
- AWG:
-
Arbitrary waveform generator
- C:
-
Capacitance
- E:
-
Electric field
- fStokes :
-
Friction
- Im[]:
-
Imaginary part
- j:
-
Imaginary indicator
- K(ω):
-
Clausius–Mossotti function
- p:
-
Polarization
- PCB:
-
Printed circuit board
- R:
-
Radius of cell (body)
- r2 :
-
Pearson's coefficient
- \(\alpha\) :
-
Confidence level
- \(\Gamma\) :
-
Torque
- \(\delta\) :
-
Membrane thickness
- \(\varepsilon\) :
-
Relative dielectric permittivity
- \({\varepsilon }_{0}\) :
-
Vacuum permittivity
- \(\eta\) :
-
Viscosity
- \(\sigma\) :
-
Conductivity
- \(\omega\) :
-
Frequency
- ΩRot :
-
Rotation velocity
- b:
-
Body
- c:
-
Cell
- corr:
-
Corrected
- cyto:
-
Cytoplasm
- exp:
-
Experimental
- m:
-
Medium
- mem:
-
Membrane
- *:
-
Complex
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Acknowledgements
The authors acknowledge Dr. Xuqu Hu for the help with the COMSOL model and Dr. Ziduo Lin and Dr. Yuqian Li for the help with lens-free image and Dr. Ying Ting Set and Dr. Lei Zhang for the discussions about DEP.
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C. D. M. C. contributed to the conceptualization of the project, methodology, performance of experiments, data analysis, validation, writing the original draft. K.T.U was responsible for the COMSOL model and correction factor results. P. B., T. M., and G. M., contributed to the implementation of electronic set-up. R.Y. contributed with the impedance matching. I. O., W. V. R., L. L and C. L. contributed to funding acquisition. C. L. and W. V. R. contributed to conceptualization, data interpretation, scientific discussions, and manuscript review.
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D. M. Campos, C., Uning, K.T., Barmuta, P. et al. Use of high frequency electrorotation to identify cytoplasmic changes in cells non-disruptively. Biomed Microdevices 25, 39 (2023). https://doi.org/10.1007/s10544-023-00677-9
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DOI: https://doi.org/10.1007/s10544-023-00677-9