Mirco Junker
last updated: March 2002
1.) Go to “Control Panels” on the Menu and click on ”Startup
Disk”.
2.) Mark the Macintosh HD:System with Mac OS 10.1.3 and restart the computer.
3.) After this start the program from the NIH Image folder and load the gel
as a .tif file.
4.) The gel must be parallel the x-axis and be shown completely. If not continue
with step 5 otherwise go to step 6.
5.) Go to Edit and rotate the picture till it is parallel. Then continue with
“Scale to fit window” out of the Options menu. Save this.
6.) Open the Special Menu and load the Macro “Gel Plotting Macros”
from Desktop (Mac OS 9) -> NIH Image -> Macros.
7.) Choose the square wave from tools and mark the first lane completely. After
it click on “Mark first Lane” under Special. You can’t change
the size of the square wave after the first lane is marked. Use this square
wave to choose the second lane and go on with “Mark Next Lane” under
Special. Repeat this till all lanes are marked. Every lane gets a number by
the computer.
8.) When all lanes are marked click on “Plot Lanes” from the Special
menu and save this data.
9.) Now all the single bands, which shall be calculated must be separated from
other bands for the computer in this plots window. So choose the icon for straight
lines out of the tool box and make a vertical line around every band you’re
interested in, so that every band has one line at the left site and one line
on the right site. If it is necessary you can also make a horizontal background
line. Try to set these lines that the whole band is inside these boxes and that
nothing from other bands is inside these boxes.
10.) If all interested lines are isolated use the “Magic Wand” from
the tool menu. With this tool every box can be calculated. Mark the boxes with
the top of this “Magic Wand”. The computer counts them automatically
so write down in which order you mark the boxes.
11.) The boxes will be calculated when you go to Analyze and click on “Show
Results”.
12.) Save this data.
13.) This file can be loaded in an Excel document.
14.) If you want to go back to the other Mac OS close all windows, open system
preferences from the menu, open startup disk and restart the computer with Mac
OS 9.2.2.