Uploaded October 2021 | Updated September 2026, 2 weeks ago
The left-hand side shows the Dodt contrast image; the right side shows the simultaneous two-photon excited fluorescence. Initially, the pipettes are positioned above the slice. They are then sequentially brought into position in front of the respective target cell and a seal formed, working from the deepest to the shallowest cell. There are several things to take particular note of. First, changes to the pressure applied to the pipette tip can be seen as changes in the dye streaming out of the respective tip. Next, dimple formation can be observed in both images for each pipette, but it is most pronounced for the third cell. Finally, the break-in to achieve whole-cell configuration is marked by the target cell beginning to fill with dye.
The left-hand side shows the Dodt contrast image; the right side shows the simultaneous two-photon excited fluorescence. Initially, the pipettes are positioned above the slice. They are then sequentially brought into position in front of the respective target cell and a seal formed, working from the deepest to the shallowest cell. There are several things to take particular note of. First, changes to the pressure applied to the pipette tip can be seen as changes in the dye streaming out of the respective tip. Next, dimple formation can be observed in both images for each pipette, but it is most pronounced for the third cell. Finally, the break-in to achieve whole-cell configuration is marked by the target cell beginning to fill with dye.
