LS2K Macro - Version b1.2

I went to Weill to troubleshoot the macro on the MRC-1024 system there. I found that the "ScanStart" command, which is needed to acquire an image, was causing all of the trouble. The program crashes when this command is called from within a .dll. However, the command works fine when it is called directly from the script editor. Since the .dll was necessary to create my GUI used in all the previous versions, I had to abandon the GUI idea (at least for now) and control the scope using just a Visual Basic script.

The user can specify the number of rows and columns of image tiles, percentage overlap between tiles, directory for saving scanned images, and COM port by changing the default values in the script. In a later version, I will try to include some kind of pop-op input box for these values to make it a little more user-friendly.

Sushmita has successfully used the script to scan and save a series of tiled images. It works well except that the images do not line up perfectly along one edge when "stitched" together. This could be due to backlash in the XY stage motor or perhaps an issue with the image stitching software. We are currently working on this issue.

The script (version b1.2) and instructions on how to use it can be found here.

Note: The "ScanStart" command does work from within a .dll on our Radiance system, so it is possible to use a GUI-based macro for acquiring images on our scope in Ithaca.

LS2KMacro – Status

The macro for the MPM system is progressing, and the current version implements 4 out of the 5 original requirements.

- From the macro GUI, the user can enter the number of columns (X direction) and rows (Y direction) to be included in the scan.

- An input box allows the user to define the degree of overlap between the frames.

- The scan can be interrupted in mid-operation by clicking the “Stop” button. Then, the user can either continue the scan from the current position by clicking the “Resume” button, or the “Start” button can be used to start a new scan from the beginning. If the user-defined settings such as X and Y frames or overlap are changed, these new settings will be used the next time Start is pressed.

- A check-box is used to provide the option of homing back to the starting position at the end of the run.

This version of the code has been tested on our system using LaserSharp2000. The features described above worked successfully, and we were able to view the images scanned by the MPM after each movement of the XY stage to the next frame.

This macro code and installation instructions can be found at http://dev.drbio.cornell.edu/lt222/LS2Kmacro/downloads.html.
Look for files under the heading "a.92". Please note that some features of the GUI are for development purposes only (such as the listbox and cheesy graphic), and this will not be the final format of the interface. Any comments/suggestions about features wanted in the final format would definitely be appreciated!

As for the next step… I am currently working on automatically saving the image files directly from the macro. I am also working on a method for taking z-stacks of selected frames. I have been developing one possible method for selecting the frames graphically. This involves overlaying a grid (each square represents one frame) on an image of the entire field of view, and the user can click on each frame they want a z-stack of. Another possible method would be selecting the frames numerically. I am also getting input from MPM users on methods they might like. (Again, any comments would be appreciated.) Another feature I will try to add in future versions is the option to move the stage to a user-selected position at the end of the scan.

-Lisa

LS2KMacro - First Test

The BioRad Radiance system is supplied with either Marzhauser or Ludl motorized XY stage. So it seems that it works with either one or the other out of the box. We'll need our macro to communicate with a different stage controller (ASI) with its own command set and firmware altogether. As a first step, we need to validate a basic requirement; make sure the LS2K macro can control the XY stage via RS232. A VB macro has been created to do just that. If everything works, it will display a simple user interface and the user should be able to perform basic stage movements with the GUI. The following link has the ActiveX DLL component and other necessary information to conduct the test. Look for version a1.0

http://dev.drbio.cornell.edu/pl47/downloads.html

-Paul