Confocal images were acquired using a Zeiss LSM 780. Labeled cells were excited at 488, 561, and 633 nm for citrine, the TMR-HaloTag ligand, and the SiR–SNAP-tag ligand, respectively. All channels were acquired with different tracks to eliminate cross-talk. A 40× objective [numerical aperture (NA), 1.2] was used to acquire images. IRM images were obtained using the same instrument with a 488-nm laser line for excitation and 470 to 505 nm for detection in the transmission channel. Values for individual cell surface areas were determined from IRM images by measuring the number of pixels corresponding to thresholded images of each cell using the tracing tool in the ImageJ software. This was converted to square micrometers by reference to pixel size.

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