Analyzing electrophysiology data

Analyzing electrophysiology data (primary analysis)

Analyzing Electrophysiology Data (Video)

1. Open pClamp Conversion v. 3 (the green frog icon on the desktop).

2. Click browse and navigate to the TEVC-8-data folder. Locate the folder corresponding to the date desired and double click the first data file in your set. Click Process.

3. pClamp will generate an Excel spreadsheet with the data collected from the experiment. By default the active sheet will have a table with all the ramps (readings) and other information. Insert a bottom borderline all the way across under the last ramp of each solution to separate the each time intervals during which different solutions were applied. To do this, look for the Bottom Border (box with a bold black line) in the Font box, on the ribbon menu.

4. Under the table heading Soln type the name of the solution corresponding to the different time periods.

5. On the Excel sheet, find the ramp number of the last ramp in every solution. Go back to the pClamp window. Under Available Ramps, select those ramps by clicking on the tick box next to them. Once all ramps have been selected, click Add. In the textbox that appears, type the name of the solutions that correspond to the ramps selected, in order and separated by commas. Click Create Plots.

NOTE: Some experiments will require ramps to be plotted in a manner different than above.

NOTE: If the experiment has a lot of ramps, you may split them into multiple plots. To do this, select a group of ramps, click Add, label the ramps, and repeat with the rest of the ramps.

6. A new spreadsheet appears with different graphs. Hover the mouse over the Current vs. Voltage graph and right click it. Click Select Data.

7. Click on the first item in the left white box and click Edit. For both the X and Y axes boxes, change the numbers after each dollar sign ($) to 150 and 1950 (e.g.=_________$150______$1950). Do NOT use the arrow keys! If you use them, the entire data range will shift and you will have to start over. Repeat for each ramp (R item in the white box) on all Current vs. Voltage graphs available.

8. At the top of the window, find and click the Insert tab. In the Illustrations box, click Shapes and select a simple line. Left click, hold and drag to insert a line vertically through the Conductance vs. Time curve in between data points where the solution was changed. Repeat this for all solution changes.

9. Look for the Text box under the Insert tab. Click Text box and then click once above the Conductance vs. Time to insert a text box. Type in the name of the first solution applied and place it over the part of the curve where the solution was used. Repeat this for all solutions.

10. In the white space under the Conductance vs. Time graph, type the file name (from the top of the Excel page) in one cell, skip a cell to the right, and then type the name and concentration of the RNA injected into the oocyte, as well as any other compounds injected into the oocyte. Move a few cells down and type in Injected by followed by the initials of the person who injected the oocyte and the date of injection.

11. Click Save. The data will automatically be saved in the Processed pClamp files folder In Local Disk C.

12. Enter the experiment into the Experiment Database, which is located in a folder on the desktop.

13. Copy the Excel file to a flash drive and plug into a computer with G: drive access. Save under the respective project folder under electrophysiology data in the unsorted data folder.

Analyzing electrophysiology data

Analyzing electrophysiology data (primary analysis)

Analyzing electrophysiology data (secondary analysis)