This paper presents a cellular force measurement technique that allows for mechanical characterization of mouse oocytes during microinjection (i.e., in situ) without requiring a separate characterization process. The technique employs an elastic cell holding device and a sub-pixel computer vision tracking algorithm to resolve cellular forces in real time with a nanonewton force measurement resolution (2 nN at 30 Hz). Mechanical properties (i.e., stiffness) of both healthy and defective mouse oocytes are characterized. The experimental results suggest that the in situ obtained force-deformation data are useful for distinguishing healthy mouse oocytes from those with aging-induced cellular defects, promising an approach for oocyte quality assessment during microinjection. Biomembrane and cytoskeleton structures of the healthy and defective oocytes are also investigated in an attempt to correlate the measured subtle mechanical difference to cellular structure changes.