Probe Sensitivity to Cortical versus Intracellular Cytoskeletal Network Stiffness

Amir Vahabikashi, Chan Young Park, Kristin Perkumas, Zhiguo Zhang, Emily K. Deurloo, Huayin Wu, David A. Weitz, W. Daniel Stamer, Robert Goldman, Jeffrey J. Fredberg, Mark Johnson*

*Corresponding author for this work

Research output: Contribution to journalArticle

Abstract

In development, wound healing, and pathology, cell biomechanical properties are increasingly recognized as being of central importance. To measure these properties, experimental probes of various types have been developed, but how each probe reflects the properties of heterogeneous cell regions has remained obscure. To better understand differences attributable to the probe technology, as well as to define the relative sensitivity of each probe to different cellular structures, here we took a comprehensive approach. We studied two cell types—Schlemm's canal endothelial cells and mouse embryonic fibroblasts (MEFs)—using four different probe technologies: 1) atomic force microscopy (AFM) with sharp tip, 2) AFM with round tip, 3) optical magnetic twisting cytometry (OMTC), and 4) traction microscopy (TM). Perturbation of Schlemm's canal cells with dexamethasone treatment, α-actinin overexpression, or RhoA overexpression caused increases in traction reported by TM and stiffness reported by sharp-tip AFM as compared to corresponding controls. By contrast, under these same experimental conditions, stiffness reported by round-tip AFM and by OMTC indicated little change. Knockout (KO) of vimentin in MEFs caused a diminution of traction reported by TM, as well as stiffness reported by sharp-tip and round-tip AFM. However, stiffness reported by OMTC in vimentin-KO MEFs was greater than in wild type. Finite-element analysis demonstrated that this paradoxical OMTC result in vimentin-KO MEFs could be attributed to reduced cell thickness. Our results also suggest that vimentin contributes not only to intracellular network stiffness but also cortex stiffness. Taken together, this evidence suggests that AFM sharp tip and TM emphasize properties of the actin-rich shell of the cell, whereas round-tip AFM and OMTC emphasize those of the noncortical intracellular network.

Original languageEnglish (US)
Pages (from-to)518-529
Number of pages12
JournalBiophysical Journal
Volume116
Issue number3
DOIs
StatePublished - Feb 5 2019

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Atomic Force Microscopy
Traction
Vimentin
Microscopy
Fibroblasts
Knockout Mice
Technology
Actinin
Finite Element Analysis
Cellular Structures
Wound Healing
Dexamethasone
Actins
Endothelial Cells
Pathology

ASJC Scopus subject areas

  • Biophysics

Cite this

Vahabikashi, A., Park, C. Y., Perkumas, K., Zhang, Z., Deurloo, E. K., Wu, H., ... Johnson, M. (2019). Probe Sensitivity to Cortical versus Intracellular Cytoskeletal Network Stiffness. Biophysical Journal, 116(3), 518-529. https://doi.org/10.1016/j.bpj.2018.12.021
Vahabikashi, Amir ; Park, Chan Young ; Perkumas, Kristin ; Zhang, Zhiguo ; Deurloo, Emily K. ; Wu, Huayin ; Weitz, David A. ; Stamer, W. Daniel ; Goldman, Robert ; Fredberg, Jeffrey J. ; Johnson, Mark. / Probe Sensitivity to Cortical versus Intracellular Cytoskeletal Network Stiffness. In: Biophysical Journal. 2019 ; Vol. 116, No. 3. pp. 518-529.
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Vahabikashi, A, Park, CY, Perkumas, K, Zhang, Z, Deurloo, EK, Wu, H, Weitz, DA, Stamer, WD, Goldman, R, Fredberg, JJ & Johnson, M 2019, 'Probe Sensitivity to Cortical versus Intracellular Cytoskeletal Network Stiffness', Biophysical Journal, vol. 116, no. 3, pp. 518-529. https://doi.org/10.1016/j.bpj.2018.12.021

Probe Sensitivity to Cortical versus Intracellular Cytoskeletal Network Stiffness. / Vahabikashi, Amir; Park, Chan Young; Perkumas, Kristin; Zhang, Zhiguo; Deurloo, Emily K.; Wu, Huayin; Weitz, David A.; Stamer, W. Daniel; Goldman, Robert; Fredberg, Jeffrey J.; Johnson, Mark.

In: Biophysical Journal, Vol. 116, No. 3, 05.02.2019, p. 518-529.

Research output: Contribution to journalArticle

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AU - Vahabikashi, Amir

AU - Park, Chan Young

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AU - Wu, Huayin

AU - Weitz, David A.

AU - Stamer, W. Daniel

AU - Goldman, Robert

AU - Fredberg, Jeffrey J.

AU - Johnson, Mark

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Vahabikashi A, Park CY, Perkumas K, Zhang Z, Deurloo EK, Wu H et al. Probe Sensitivity to Cortical versus Intracellular Cytoskeletal Network Stiffness. Biophysical Journal. 2019 Feb 5;116(3):518-529. https://doi.org/10.1016/j.bpj.2018.12.021