Table 2:

Troubleshooting Table

Step	Problem	Possible reason	Solution
7	Wells are not properly positioned and walls are not perpendicular	Biopsy punches were not properly used	Print a schematic of the design and use it as a guide. Pay attention to applying pressure perpendicularly to the surface
8	The 1.5 mm diameter connecting channel is difficult to punch	There is not sufficient structural integrity to pierce the PDMS	Place the cored 8-mm plug back inside the well to absorb the pressure of the 1.5-mm punch
28	Germaria are bunched up with egg chambers in the muscle sheath	The muscle sheath was intact. It should be broken/ ripped near the stage 7 egg chamber.	During dissection pinch off stage 7/8 and older egg chambers from the ovariole with forceps.
41	Images become darker over time in transmitted light channel Imaging software cannot handle the large dataset being acquired	An air bubble formed just below the chamber lid Images are being saved to memory	Make two 1-mm holes in the PDMS lid above the imaging well Use settings in imaging software to autosave to hard drive instead