The injection of positive charge carriers (holes) into a single conjugated polymer chain was observed to be light-assisted. This effect may underlie critical, poorly understood organic electronic device phenomena such as the build-up of functional deeply trapped charge layers in polymer light emitting diodes. The charging/ discharging dynamics were investigated indirectly by a variety of single molecule electro-optical spectroscopic techniques, including an ''image-capture'' approach.nanoparticle ͉ single molecule spectroscopy ͉ charge injection