Source: Jones, D. P., et al. Longitudinal Measurement of Extracellular Matrix Rigidity in 3D Tumor Models Using Particle-tracking Microrheology. J. Vis. Exp. (2014).
This protocol describes a technique for monitoring extracellular matrix rigidity changes by using particle-tracking microrheology (PTM). This is done by culturing 3D pancreatic tumor spheroids into ECM with embedded fluorescent probes for video particle-tracking and analysis.
1. Preparing 3D Tumor Spheroids Embedded in ECM
2. Construct Grid of Sample Points and Take a Video at Each Point
The authors have nothing to disclose.
Bovine type 1 collagen | BD Biosciences, San Jose, CA | 354231 | |
PANC-1 | American Type Cell Culture, Manassas, VA | CRL1469 | or other appropriate cell type |
Fluorescent Microspheres | Life Technologies, Carlsbad, CA | 906906 | |
Agarose | Fisher Bioreagents, Waltham, MA | C12H18O9 | |
NaOH | Fisher Bioreagents, Waltham, MA | NC0480985 | |
96-well Imaging plates | Corning Inc., Corning, NY | 3904 | |
DMEM | Hyclone, Waltham, MA | SH30243.01 | or appropriate cell culture media |
Zeiss AxioObsever Microscope | Zeiss, Oberkochen, Germany | includes high-speed camera and imaging software |