Limitations of Qdot labelling compared to directly-conjugated probes for single particle tracking of B cell receptor mobility.

ABSTRACT: Single-particle tracking (SPT) is a powerful method for exploring single-molecule dynamics in living cells with nanoscale spatiotemporal resolution. Photostability and bright fluorescence make quantum dots (Qdots) a popular choice for SPT. However, their large size could potentially alter the mobility of the molecule of interest. To test this, we labelled B cell receptors on the surface of B-lymphocytes with monovalent Fab fragments of antibodies that were either linked to Qdots via streptavidin or directly conjugated to the small organic fluorophore Cy3. Imaging of receptor mobility by total internal reflection fluorescence microscopy (TIRFM), followed by quantitative single-molecule diffusion and confinement analysis, definitively showed that Qdots sterically hinder lateral mobility rega

SUBMITTER: Abraham L 

PROVIDER: S-EPMC5595841 | biostudies-literature | 2017 Sep

REPOSITORIES: biostudies-literature