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A DNA-based molecular probe for optically reporting cellular traction forces.

A DNA-based molecular probe for optically reporting cellular traction forces.

Nature methods (2014-10-13)

Brandon L Blakely, Christoph E Dumelin, Britta Trappmann, Lynn M McGregor, Colin K Choi, Peter C Anthony, Van K Duesterberg, Brendon M Baker, Steven M Block, David R Liu, Christopher S Chen

초록

We developed molecular tension probes (TPs) that report traction forces of adherent cells with high spatial resolution, can in principle be linked to virtually any surface, and obviate monitoring deformations of elastic substrates. TPs consist of DNA hairpins conjugated to fluorophore-quencher pairs that unfold and fluoresce when subjected to specific forces. We applied TPs to reveal that cellular traction forces are heterogeneous within focal adhesions and localized at their distal edges.

MATERIALS

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Sigma-Aldrich

Hydrocortisone, ≥98% (HPLC)

USP

Hydrocortisone, United States Pharmacopeia (USP) Reference Standard

Sigma-Aldrich

Hydrocortisone, meets USP testing specifications

Sigma-Aldrich

Methanol, HPLC Plus, ≥99.9%

Sigma-Aldrich

Methanol, puriss., meets analytical specification of Ph Eur, ≥99.7% (GC)

Sigma-Aldrich

Methanol, ACS reagent, ≥99.8%

Sigma-Aldrich

Methanol, ACS reagent, ≥99.8%

Sigma-Aldrich

Methanol, BioReagent, ≥99.93%

Supelco

Methanol, analytical standard

Sigma-Aldrich

Methanol, puriss. p.a., ACS reagent, reag. ISO, reag. Ph. Eur., ≥99.8% (GC)

Sigma-Aldrich

Methanol, anhydrous, 99.8%

Supelco

Methanol, Pharmaceutical Secondary Standard; Certified Reference Material

Sigma-Aldrich

Methanol, ACS spectrophotometric grade, ≥99.9%

Sigma-Aldrich

Methanol, ACS reagent, ≥99.8%

Sigma-Aldrich

Methanol, suitable for HPLC, ≥99.9%

Sigma-Aldrich

Methanol, Laboratory Reagent, ≥99.6%

Sigma-Aldrich

Magnesium chloride solution, 1.00 M±0.01 M

Sigma-Aldrich

Magnesium chloride solution, BioUltra, ~1 M in H₂O

Sigma-Aldrich

Magnesium chloride solution, BioUltra, ~0.025 M in H₂O

Sigma-Aldrich

Magnesium chloride solution, BioUltra, 2 M in H₂O

Sigma-Aldrich

Formamide, spectrophotometric grade, ≥99%

Sigma-Aldrich

Formamide, BioUltra, ≥99.5% (T)

Sigma-Aldrich

Formamide, ReagentPlus^®, ≥99.0% (GC)

Sigma-Aldrich

Acetic acid, glacial, puriss. p.a., ACS reagent, reag. ISO, reag. Ph. Eur., ≥99.8%

Sigma-Aldrich

Hydrocortisone, γ-irradiated, powder, BioXtra, suitable for cell culture

Sigma-Aldrich

(−)-Riboflavin, BioReagent, suitable for cell culture, suitable for insect cell culture, ≥98%

Sigma-Aldrich

Hydrocortisone, BioReagent, suitable for cell culture

Sigma-Aldrich

L-Thyroxine sodium salt pentahydrate, γ-irradiated, powder, BioXtra, suitable for cell culture

Sigma-Aldrich

Zirconyl chloride octahydrate, reagent grade, 98%

Sigma-Aldrich

Urea solution, BioUltra, ~8 M in H₂O