Representation Bias, Correction Transfer, and Resolution Sensitivity in Three-Dimensional Mitochondrial Morphometry

Abstract

Quantitative imaging pipelines can produce precise but systematically different measurements of the same object. We present an empirical reliability assessment of three-dimensional mitochondrial morphometry that connects representation bias, a controlled processing intervention, correction transfer, and resolution sensitivity. Using 2,720 development objects from the 3D Mitochondria Shape Library for Optical Microscopy, we find that occupancy-derived volumes exceed reference mesh volumes by 3.665% on average despite an intraclass correlation coefficient of 0.994. Boundary analysis identifies an outward label displacement of 0.00304 normalized units. In a controlled label-pipeline reimplementation, removing the depth offset reduces volume error in all 55 analyzed objects by a mean of 1.57 percentage points, approximately 45% of mean reproduced inflation; the source of the remainder is not isolated. A frozen regression using occupancy-derived features reduces median absolute percentage error from 3.481% to 0.664% in 2,728 previously unused objects from the same resource. However, its calibrated error bound covers only 92.1% overall and 49.2% in a low-occupancy subgroup, demonstrating that accuracy and uncertainty transfer must be evaluated separately. In 550 rat-cortex objects from the MitoEM resource, coarsening in-plane spacing from 8 to 24 nanometers changes median surface area by minus 10.60% and sphericity by plus 11.76%, despite a rank correlation of 0.994. These results provide quantitative checks for distinguishing processing-induced descriptor changes from candidate biological differences, without establishing biological invariance or cross-source correction transfer.