Figure 2. Alignment of the bulk loaded neurons in the TPLSM with the cytochrome oxidase barrel staining using biocytin electroporation of single neurons.
(A) TPLSM Fluo-4 bulk loaded image of the imaged field. At the end of the recording session single cell electroporation of biocytin was performed on the bulk loaded neurons (typically, 1–4 calcium loaded neurons). Yellow arrow point to the one of the electroporated cells (right panel). These biocytin electroporated neurons served as a definite marker to the location of the calcium bulk staining. (B) Brains were fixed and sliced (100 µm thick) at the tangential plane, and barrels were visualized according to the cytochrome oxidase dense regions typical to layer 4 barrels. Alignment of the neurons labeled with biocytin (in cortical layer 2–3) to the layer 4 cytochrome oxidase stained barrel according to identified ascending blood vessels (designated by numbers 1–7) in three consecutive slices from layer 2–3 to layer 4 (slices: 100–200 µm; 200–300 µm; 400–500 µm). (C) Superposition of the biocytin-labeled neurons onto the identified barrel in layer 4 (C2 in this example). (D) The biocytin-labeled neurons are shown in higher magnification (arrow point to the identified cell in A).
Figure 2—figure supplement 1. Alignment of the bulk loaded neurons onto the barrel using single elecrtoporated cells.
