Multicolor BaLM. (A) BaLM reconstruction showing the localization of microtubules (purple), clathrin pits (yellow), and myosin IIA filaments (blue) in the same cell. Immunofluorescent labeling of myosin IIA, clathrin light chain, and microtubules was performed with Alexa Fluor 647, Alexa Fluor 488, and Alexa Fluor 561, respectively. The channels were imaged in the following order: Alexa Fluor 647, Alexa Fluor 488, and Alexa Fluor 561. (B) The diffraction limited images (Left) and the BaLM images (Right) of a single microtubule that has been labeled with a mixture of Alexa Fluor 405 (purple), Alexa Fluor 488 (green), Alexa Fluor 561 (orange), and Alexa Fluor 647 (red) secondary antibodies is shown. The channels were imaged in the following order: Alexa Fluor 647, Alexa Fluor 405, Alexa Fluor 488, and Alexa Fluor 561. (Scale bars: 2 μm.)