二抗物种特异性对双色storm成像的影响（特邀）

Abstract

Stochastic optical reconstruction microscopy (STORM), a superresolution imaging technology based on immunofluorescence, has gained popularity owing to its straightforward principle, simple optical path, and excellent spatial resolution. However, the enhanced resolution demands greater specificity from antibodies. While direct labeling with primary antibodies is an option, indirect labeling using a "primary + secondary antibody" combination is more commonly employed in practical applications. Considering the issue of species specificity with secondary antibodies, preadsorption is required to increase specificity during production. In this study, we aim to explore the effect of secondary antibody species specificity on dualcolor STORM imaging. In the classical erythrocyte skeleton model, based on the mutually exclusive localization between the Nterminal and Cterminal of βspectrin, we performed twocolor STORM imaging of them by labeling them with lowand highadsorption secondary antibodies, respectively. A comparison with crosscorrelation data from simulations revealed that lowadsorption secondary antibodies led to colocalization artifacts in