Fig. 2.

Experimental overview of our targeted AT iLEM approach. a sample preparation. The tissue block is sliced on an ultramicrotome and mounted on the sample holder using an ITO coverslip. b LM acquisition. A transmitted light source was added to generate an overview image consisting of a mosaic of transmitted light images using a 20x optical lens. Additional 100x imaging is applied. LM imaging is carried out at ambient pressure. c EM acquisition. A JEOL SJF7200 SEM retrofitted with the commercial SECOM platform was used. d introduces key steps of the workflow. The workflow can be divided into (i) preparation (overview image and focus map creation), (ii) processing (ribbon detection, section recognition, normalization of section geometries for the prediction of the position of ROIs in consecutive sections), (iii) automated navigation and image acquisition (with increasing magnifications), and (iv) stepwise refinement (using a feedback mechanism for the navigation based on the image alignment at each magnification step). For more information on the workflow, see Fig. 1