Software Process

1. System Controlling Software

CSLM controller is a custom-programmed executable software. It is originally programmed with National Instruments Labview 7. It can be used to generate the driving waveform for the galvanometer, and digitize the voltage signal from the photodetector. The "Amplitude" indicates the maximum deflect angle for the galvos, thus it decides the filed of view. The "Matrix size" and "Frame rate" decide the collection size and speed of the frame. Z-axis motor is controlled via the setup of "Step size", "Up", and "Down". The images can be adjusted for proper gain and offset and be shown and store in real-time.



Click HERE to Download the free CSLM Controller

 

2. Image Processing

ImageJ is a very powerful image processing software. Here are some applications to use it to eliminate the background, generate the 3-D projection, rendering the image sequence with color, and save the result into Windows AVI format.

First, click on “File-open” to open the background image collected when the shutter of the microscope is closed. (That way the reflection from the surface of the optical elements are collected). Then, click on “File-Import-Image Sequence” to upload the image sequence. By choosing “Process-Image Calculator”, and choose the subtract operation as follows, the background-subtracted image sequence are created.

The 3D projection can be doen with “Image-Stacks-3D Projection”.

To enhance the visual effect of the movie, click on “Image-Type-RGB Color” to convert the 8-bit grayscale image sequences to RGB format. Then, the color balance adjustment function finishes the rendering of the image sequence. Choosing “Image-Adjust-Color Balance”, and click on “Auto” or to adjust the Minimum, Maximum, and Brightness values manually, the images can be enhanced.

Applying the enhancing to all stacks:

After that, save the file to AVI format so that it can be converted to QuickTime Movie format with Ulead GIF Animator.

 

Ulead GIF Animator can automatically create the color palette to compress the movie file size. By clicking “Optimize”, the compressed movie is generated.

Save the file to QuickTime Movie Format, and choosing “Options-Compression”, choosing “Video” in “Compression” and then click “OK”. Your movie is finally done!