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. Author manuscript; available in PMC: 2018 Jan 1.
Published in final edited form as: Genesis. 2017 Jan;55(1-2):10.1002/dvg.23004. doi: 10.1002/dvg.23004

Figure 1. Method to simultaneously apply a reproducible strain and analyse cell division in the Xenopus animal cap.

Figure 1

a) The animal cap is dissected (red lines) from an early gastrula stage embryo that has been labeled by injection of GFP-α-tubulin and Cherry-histone2B mRNAs at the 2 cell stage. The dark brown area indicates the pigmented apical cell layer that will be imaged. b) Animal caps are cultured on a fibronectin coated flexible PDMS membrane until adhered. The PDMS membrane is cast in a custom mould with teeth on all four sides allowing either a uni-axial or biaxial stretch to be applied. c) PDMS membrane with cultured animal caps (arrow) attached to stretch apparatus. A uniaxial stretch is applied to stretch the animal caps by 35%; the teeth orthogonal to the stretch direction have been removed. d) The stretch apparatus is attached to the stage of an upright confocal microscope; a water-dipping objective is used to visualize cells in the apical layer. e) Example timelapse series using maximum intensity projections of cell division in the apical cell layer with frames 5 minutes apart. 35% stretch was applied in horizontal direction. The dividing cell is marked by dashed outline at 0 min, GFP-α-tubulin (green) labels the microtubule cytoskeleton and condensing centrosomes (arrowheads), Cherry-histone2B (magenta) labels nuclei. Scale bar: 30μm.