Setting up a SICM

There is more than one way to set up a SICM. behindLight was developed to control a SICM that consists of the following (products marked with an asterisk are not available commercially any more, but similar models are):

• 2 CIO-DAC02/16 cards (Measurement computing)(manufacturer's catalogue entry)
• 1 CIO-DAS802/16 card (Measurement computing)(manufacturer's catalogue entry)
• 1 Nanocube® P-611.3S (PI)(manufacturer's catalogue entry)
• 1 E-664 NanoCube® Piezo Controller (PI)(manufacturer's catalogue entry)
• 1 inverted microscope, Axiovert S100^* (Zeiss) with object translation stage, suitable for 3.5 cm diameter cell culture dishes
• 1 current clamp amplifier, 2 channels^* (Jens Meyer electronics)
• 3 M-105 Translation Stages (PI) (manufacturer's catalogue entry)
• 1 IBM compatible PC, 2 ISA-slots must be available on the motherboard, since the cards above are ISA-cards, operating system Windows 2000
• 1 Oscilloscope HM-205-3^* (Hameg)
• additional material such as screws, conductors, etc.

Fig. 1: Drawing of the amplifier's headstage with electrode holder mounted on the nanocube®.

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The amplifier's head is mounted on the Nanocube® via a perspex holder and has a fixed holding device for the glass electrode (see drawing in figure one). From now on, if in this text, the phrase "front of the Nanocube®" refers to this side. The Nanocube® as well as the amplifier's headstage are connected to their corresponding amplifiers.

Fig 2: Drawing of the mounting of Nanocube® and translation stages to make the electrode adjustable in all three dimensions (structure shown in figure one is not shown for clarity)

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The back of the Nanocube® is attached to one of the M-105 translation stages. The heading direction of the stage should be parallel to the glass electrode, it will be used to position the electrode manually in z-direction.

This z-translation stage is mounted perpendicularly via a L-formed aluminium device on a second translation stage, that is itself mounted perpendicularly on the third translation stage. Via these three stages the Nanocube® and thus the attached glass pipette can be positioned manually in all three dimensions. The drawing in figure 2 shows the nanocube and the three stages. To keep the drawing comprehensible, the holder of the amplifier's head and the electrode holder have been omitted.

Fig. 3: Drawing of the translation stages mounted to the microscope's object translation stage.

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The lowest translation stage is attached to a aluminium stage that is mounted on a column standing perpendicularly on the microscope's object translation table. In figure 3, again, for clarity only the lowest translation table is drawn, the structures from figure 1 and 2 are not shown.

The complete construction described above should be easily tilted to allow one to change the cell culture dishes on the miscroscope's stage. Of course, the dimensions of the construct should be suitable for manually positioning the scanning sample and the glass electrode. The tip of the glass electrode has to be detectable in the field of view of the microscope. To perform a successful scan, the tip of the glass electrodes must be manually positionable in the z-direction such that the tip of the glass electrode and the sample have a maximal distance of 100 micrometer (the maximum extension of the nanocube®).

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