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Description of patch clamp electrode drawing manual for technology sharing

Time:2021-04-15 Views:231

After you understand what kind of electrode is a good electrode, the next step is how to use P-97 to make the ideal electrode.

First, set the pressure to 500 (generally the default is 500), and then use different parameters for different platinum plates. The trapezoidal platinum plate is commonly used in my laboratory, and there is also a square platinum plate. First use the RAMP function to measure the melting point of the glass tube (this melting point is represented by RAMP). The parameter list corresponding to the two platinum plates is as follows:

Platinum plate type

HEAT

PULL

VELOCITY

TIME

Trapezoid

RAMP+15

0

40

200

Square

RAMP

0

40

200

   First use the above parameter settings for one step. Start drawing! It is often impossible to break the electrode in such a step, so P-97 will automatically execute the program in a loop until the electrode is broken. For example, at this time, the drawing procedure is looped three times, that is, the electrode is broken in three steps. Then use a polisher (or microscope) to observe the click tip. The electrode that is broken in three steps is often thinner and the tip is steep and the opening is too small. Such an electrode is not ideal. Next, we only adjust the drawing speed, reducing the drawing speed can make the click fatter. You can reduce the drawing speed step by step by "3" and you can make the following records. E.g:

Drawing speed

40

37

34

31

28

25

22

Number of drawing steps

3

3

4

4

4

4

5

You will find that the shapes of the electrodes pulled off with the same number of steps are similar, but the difference between the number of asynchronous steps is relatively large. Generally speaking, a four-step broken electrode is ideal. In the above example, you should not set the drawing speed to 34 or 25 because they are at the critical value of step change. Since the drawing instrument has some fluctuations every time it is drawn, if you set the critical value, it is very likely that the number of drawing steps will change during the drawing process and the drawing will fail. You should set the straightening speed to 31 or 28 so that it will be more stable.

If this is the ideal electrode that has been drawn, then the setting of the drawing instrument ends here, and you can use this parameter to draw in the future. If you find that the clicks made with four steps are too thin, but the clicks made with five steps are too thick, it means that each drawing without using the same parameters can no longer meet your requirements. At this time, you need to use different parameter settings. It is like thinking that the four steps are too thin and the five steps are too thick in the above situation. First, you have to set the parameters of the four steps in a program (assuming RAMP=285 using trapezoidal platinum plates) as follows:

HEAT

PULL

VELOCITY

TIME

300

0

28

200

300

0

28

200

300

0

28

200

300

0

23

200

It should be noted that special attention needs to be paid to the last place to be completed. It is necessary to appropriately reduce the speed of the last step to pull out a fatter ideal electrode.


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