<p>Micro-Manager- Multi-D Acquire for Time Lapse Imaging</p><p>1. Open Micro-Manager 2. Turn on LED light source and determine binning, exposure, gain and bit level for your sample. 3. Click on Multi-D acquire- you will get this new window</p><p>4. In the Time Points box a. Set the total number of frames you want to acquire b. Set the interval between frames and the units (seconds or minutes) 5. In the Channels box a. If there is nothing, choose channels from the pull down and then New b. In the Configuration pick the channel you want (White, Blue, Green) c. Set the exposure to what you determined you need in step 2 d. If you are doing two channels, add New for another channel and set the exposure 6. Save Images- this determines what will happen to the images when they are captured a. If it is unchecked then the images will be stored in memory. This is good for playing around as you learn and you start and stop several times to get it right. As you close each window, the data is lost. If you want to save, you can always do that before closing. b. If Save Images is checked, the images will be saved to the hard drive. The location is determined in the Directory box. The button with (…) will allow you to navigate and set the location. It will default to where you last saved. It will create a new folder each time you start and acquisiion, so if you point it at your folder, all the data will come up there in folders i. If you choose Display: All then all of the images you capture can be seen and played as you are running the experiment or when done. But, if you run out of system memory, the program will stop acquiring. You will not lose your data, but you will not collect any more. So if you are doing a long experiment (overnight) this is dangerous) ii. If you choose Display: Last then you will see each frame as it is acquired, but will not be able to view previous frames until you are done and load the whole movie. This is the mode to use for long experiments.</p>