AFM and Fluorescence Microscopy of Single Cells with Simultaneous Mechanical Stimulation via Electrically Stretchable Substrates
We have developed a novel experimental set-up that simultaneously, (i) applies static and dynamic deformations to adherent cells in culture, (ii) allows the visualization of cells under fluorescence microscopy, and (iii) allows atomic force microscopy nanoindentation measurements of the mechanical p...
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2021-07-01
|
| Series: | Materials |
| Subjects: | |
| Online Access: | https://www.mdpi.com/1996-1944/14/15/4131 |
| _version_ | 1797525412185636864 |
|---|---|
| author | Natalia Becerra Barbara Salis Mariateresa Tedesco Susana Moreno Flores Pasquale Vena Roberto Raiteri |
| author_facet | Natalia Becerra Barbara Salis Mariateresa Tedesco Susana Moreno Flores Pasquale Vena Roberto Raiteri |
| author_sort | Natalia Becerra |
| collection | DOAJ |
| description | We have developed a novel experimental set-up that simultaneously, (i) applies static and dynamic deformations to adherent cells in culture, (ii) allows the visualization of cells under fluorescence microscopy, and (iii) allows atomic force microscopy nanoindentation measurements of the mechanical properties of the cells. The cell stretcher device relies on a dielectric elastomer film that can be electro-actuated and acts as the cell culture substrate. The shape and position of the electrodes actuating the film can be controlled by design in order to obtain specific deformations across the cell culture chamber. By using optical markers we characterized the strain fields under different electrode configurations and applied potentials. The combined setup, which includes the cell stretcher device, an atomic force microscope, and an inverted optical microscope, can assess in situ and with sub-micron spatial resolution single cell topography and elasticity, as well as ion fluxes, during the application of static deformations. Proof of performance on fibroblasts shows a reproducible increase in the average cell elastic modulus as a response to applied uniaxial stretch of just 4%. Additionally, high resolution topography and elasticity maps on a single fibroblast can be acquired while the cell is deformed, providing evidence of long-term instrumental stability. This study provides a proof-of-concept of a novel platform that allows in situ and real time investigation of single cell mechano-transduction phenomena with sub-cellular spatial resolution. |
| first_indexed | 2024-03-10T09:13:27Z |
| format | Article |
| id | doaj.art-de072057b78c4275896100e20b434d42 |
| institution | Directory Open Access Journal |
| issn | 1996-1944 |
| language | English |
| last_indexed | 2024-03-10T09:13:27Z |
| publishDate | 2021-07-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Materials |
| spelling | doaj.art-de072057b78c4275896100e20b434d422023-11-22T05:52:11ZengMDPI AGMaterials1996-19442021-07-011415413110.3390/ma14154131AFM and Fluorescence Microscopy of Single Cells with Simultaneous Mechanical Stimulation via Electrically Stretchable SubstratesNatalia Becerra0Barbara Salis1Mariateresa Tedesco2Susana Moreno Flores3Pasquale Vena4Roberto Raiteri5Department of Informatics, Bioengineering, Robotics, and System Engineering (DIBRIS), University of Genova, 16145 Genova, ItalyDepartment of Informatics, Bioengineering, Robotics, and System Engineering (DIBRIS), University of Genova, 16145 Genova, ItalyDepartment of Informatics, Bioengineering, Robotics, and System Engineering (DIBRIS), University of Genova, 16145 Genova, ItalyIndependent Researcher, 1190 Vienna, AustriaDepartment of Chemistry, Materials and Chemical Engineering Giulio Natta, Politecnico di Milano, 20133 Milan, ItalyDepartment of Informatics, Bioengineering, Robotics, and System Engineering (DIBRIS), University of Genova, 16145 Genova, ItalyWe have developed a novel experimental set-up that simultaneously, (i) applies static and dynamic deformations to adherent cells in culture, (ii) allows the visualization of cells under fluorescence microscopy, and (iii) allows atomic force microscopy nanoindentation measurements of the mechanical properties of the cells. The cell stretcher device relies on a dielectric elastomer film that can be electro-actuated and acts as the cell culture substrate. The shape and position of the electrodes actuating the film can be controlled by design in order to obtain specific deformations across the cell culture chamber. By using optical markers we characterized the strain fields under different electrode configurations and applied potentials. The combined setup, which includes the cell stretcher device, an atomic force microscope, and an inverted optical microscope, can assess in situ and with sub-micron spatial resolution single cell topography and elasticity, as well as ion fluxes, during the application of static deformations. Proof of performance on fibroblasts shows a reproducible increase in the average cell elastic modulus as a response to applied uniaxial stretch of just 4%. Additionally, high resolution topography and elasticity maps on a single fibroblast can be acquired while the cell is deformed, providing evidence of long-term instrumental stability. This study provides a proof-of-concept of a novel platform that allows in situ and real time investigation of single cell mechano-transduction phenomena with sub-cellular spatial resolution.https://www.mdpi.com/1996-1944/14/15/4131cell stretchingmechanical stimulationdielectric elastomer actuatorsatomic force microscopycellular biomechanics |
| spellingShingle | Natalia Becerra Barbara Salis Mariateresa Tedesco Susana Moreno Flores Pasquale Vena Roberto Raiteri AFM and Fluorescence Microscopy of Single Cells with Simultaneous Mechanical Stimulation via Electrically Stretchable Substrates Materials cell stretching mechanical stimulation dielectric elastomer actuators atomic force microscopy cellular biomechanics |
| title | AFM and Fluorescence Microscopy of Single Cells with Simultaneous Mechanical Stimulation via Electrically Stretchable Substrates |
| title_full | AFM and Fluorescence Microscopy of Single Cells with Simultaneous Mechanical Stimulation via Electrically Stretchable Substrates |
| title_fullStr | AFM and Fluorescence Microscopy of Single Cells with Simultaneous Mechanical Stimulation via Electrically Stretchable Substrates |
| title_full_unstemmed | AFM and Fluorescence Microscopy of Single Cells with Simultaneous Mechanical Stimulation via Electrically Stretchable Substrates |
| title_short | AFM and Fluorescence Microscopy of Single Cells with Simultaneous Mechanical Stimulation via Electrically Stretchable Substrates |
| title_sort | afm and fluorescence microscopy of single cells with simultaneous mechanical stimulation via electrically stretchable substrates |
| topic | cell stretching mechanical stimulation dielectric elastomer actuators atomic force microscopy cellular biomechanics |
| url | https://www.mdpi.com/1996-1944/14/15/4131 |
| work_keys_str_mv | AT nataliabecerra afmandfluorescencemicroscopyofsinglecellswithsimultaneousmechanicalstimulationviaelectricallystretchablesubstrates AT barbarasalis afmandfluorescencemicroscopyofsinglecellswithsimultaneousmechanicalstimulationviaelectricallystretchablesubstrates AT mariateresatedesco afmandfluorescencemicroscopyofsinglecellswithsimultaneousmechanicalstimulationviaelectricallystretchablesubstrates AT susanamorenoflores afmandfluorescencemicroscopyofsinglecellswithsimultaneousmechanicalstimulationviaelectricallystretchablesubstrates AT pasqualevena afmandfluorescencemicroscopyofsinglecellswithsimultaneousmechanicalstimulationviaelectricallystretchablesubstrates AT robertoraiteri afmandfluorescencemicroscopyofsinglecellswithsimultaneousmechanicalstimulationviaelectricallystretchablesubstrates |