1 × 10^5 cells MDA-MB-231 cells were plated on rimmed petri dishes (Ted Pella, Inc) coated with 1 mg/mL unlabeled gelatin. The Young’s modulus of MDA-MB-231 cells in L15 phenol red free cell culture media (Gibco) with 10% FBS (Gemini Bio) was measured using a Flex-Bio AFM (Nanosurf AG, Switzerland) equipped with an 178 inverted microscope Axio Observer (Carl Zeiss) and an environmental control enclosure. The AFM is operated in static force mode, and with a cantilever with spring constant 0.22 N, and its silicon tip with radius <10 nm (Stat0.2LAuD, Nanosurf AG, Switzerland). Force curves were measured across cells within 30 μm windows with each data point collected every 2 μm. A custom Matlab (MathWorks, Natick, MA, USA) code was written to select and average the elasticity of 25 data points that have the highest contact points with the cell, which corresponds to the nucleus of the cell. The Sneddon model was used to approximate the Young’s modulus, with a cone with half angle 15°, fitting from 0 – 500 pN.
L15 phenol red free cell culture media
Manufactured by Thermo Fisher Scientific
Sourced in Switzerland
L15 phenol red free cell culture media is a product designed for cell culture applications. It is a basal medium that provides a nutrient-rich environment for the growth and maintenance of various cell types. The absence of phenol red in this formulation makes it suitable for applications where the presence of this pH indicator may interfere with experimental protocols or observations.
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2 protocols using l15 phenol red free cell culture media
1
Measuring Young's Modulus of MDA-MB-231 Cells
2
Measuring Young's Modulus of MDA-MB-231 Cells
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1 × 10^5 cells MDA-MB-231 cells were plated on rimmed petri dishes (Ted Pella, Inc) coated with 1 mg/mL unlabeled gelatin. The Young’s modulus of MDA-MB-231 cells in L15 phenol red free cell culture media (Gibco) with 10% FBS (Gemini Bio) was measured using a Flex-Bio AFM (Nanosurf AG, Switzerland) equipped with an 178 inverted microscope Axio Observer (Carl Zeiss) and an environmental control enclosure. The AFM is operated in static force mode, and with a cantilever with spring constant 0.22 N, and its silicon tip with radius <10 nm (Stat0.2LAuD, Nanosurf AG, Switzerland). Force curves were measured across cells within 30 μm windows with each data point collected every 2 μm. A custom Matlab (MathWorks, Natick, MA, USA) code was written to select and average the elasticity of 25 data points that have the highest contact points with the cell, which corresponds to the nucleus of the cell. The Sneddon model was used to approximate the Young’s modulus, with a cone with half angle 15°, fitting from 0 – 500 pN.
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