Biomechanics plays a central role in breast epithelial morphogenesis. In this study we have used 3D cultures in which
normal breast epithelial cells are able to organize into rounded acini and tubular ducts, the main structures found in the
breast tissue. We have identified fiber organization as a main determinant of ductal organization. While bulk rheological
properties of the matrix seem to play a negligible role in determining the proportion of acini versus ducts, local changes
may be pivotal in shape determination. As such, the ability to make microscale rheology measurements coupled with
simultaneous optical imaging in 3D cultures can be critical to assess the biomechanical factors underlying epithelial
morphogenesis. This paper describes the inclusion of optical tweezers based microrheology in a microscope that had
been designed for nonlinear optical imaging of collagen networks in ECM. We propose two microrheology methods and show preliminary results using a gelatin hydrogel and collagen/Matrigel 3D cultures containing mammary gland
epithelial cells.
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