成像分析标准化缺失：如何实现定量共定位与光谱串扰校正？

The imaging analysis lacks standardized, quantitative colocalization metrics with bleed-through correction and randomization-based significance testing. Visual overlap and line scans (Figure 1A-B) are susceptible to spectral bleed-through and local crowding; PLA foci counts (Figure 1C) can scale with mitochondrial density and cell size. Without Pearson/Manders coefficients [1][2], bleed-through correction [3], and statistical significance testing [4], the strength and specificity of spatial association remain uncertain. These represent established analytical standards rather than emerging methodologies.