Target software:
- Control multiple pipettes
- Image click moves pipettes
- Collision protection
- Focus follows electrode
- Global list
Target - Image driven 3D Micropositioning Software
Target software:
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| Robotic Targeting | Target program | Download | Acknowledgements |
"Robotic targeting" refers to a robotic software and hardware system that allows a human to direct micropipettes and other micro-tools to microscopic objects such as a living biological cells arranged in a 3D optical space. Robotic means that the computer assists in these movements making sure they are safe. Targeting means that an investigator interactively moves micropipettes or other probes in three dimensions under a microscope by clicking on a focused image of the object with a computer mouse. The software translates the user-selected 3-D object point(s) (mouse-click (x,y) and level of focus (z)) into precision targeting movements by a motorized micropositioner. Biotechnology applications include microinjection and micromanipulation of cells, electrophysiological recording, and microdelivery of pharmacological agents to cells for drug testing and diagnostics. Additional application is fabrication of microelectronics.
Robotic targeting simplifies your use of microprobes under a light microscope. It does this by simplifying a number of tedious and sometimes impossible operations. In some cases it prevents you from destroying a preparation, which is sometimes a risk with motor controlled micromanipulators. To appreciate the benefits of robotic targeting consider the following scenarios:
Mounting new electrodes necessitate inserting the electrode under a high powered objective. Robotic Targeting simplifies this in several ways: (1) allows you to click on the electrode tip under low power instead of having you move the electrode to a special rectangle drawn on a video monitor; (2) temporarily extracts all electrodes while you change the objective.
Inserting electrodes into deep tissue require drawing an electrode out of the field of view, lowering the electrode (by guesswork), and blindly inserting the electrode into the tissue. Robotic targeting simplifies this operation by allowing you to bring the desired position into focus and then click on the desired position. The electrode automatically withdraws, moves and inserts according to a variety of paths and insertion protocols.
Selecting a new tissue site for study requires moving the field of view and then moving one or more electrodes. Our Target program permits moving electrodes to the field of view, as well as moving to previous fields of view or moving the field of view and level of focus to the electrode.
Focus and Stage follows electrode while you move the electrode with the manufacturer's manual input device (joystick or other). This is valuable, for example, when dragging a cell soma from deep tissue.
The stand-alone robotic targeting application contains five windows: Control, Manual, Joy, List, and Image. The Control window provides buttons for various special features such as selecting electrode or objective, moving electrode, level of focus or field of view, removing electrodes, setting a "safe" level, retracting electrodes from tissue, extracting electrodes to change objective, moving electrode to focus or focus to electrode or field of view to electrode or electrode to field of view. There are automated "electrode following" methods. NOTE: the following picture is old. Location tools have been removed; action tools have been added; the window now has a vertical arrangement (18 Nov. 2005).
The Manual window permits you to move motors in a step-wise manner. You click on the x, y, and z columns of the manipulator keys to move the respective axis up or down. You click on the "S-" or "S+" keys to change the step size which is reflected in the Target Control window. Keys on the computer keyboard duplicate functionality of this window. These short-cuts speed the process of moving motors is a successively refined manner. The step size is changed by factors of 2 or 2.5 so that it is always some power of ten times 1, 2 or 5. |
The Joy window applies to those motor controllers that have joy sticks, and for which the joy-stick sensitivity is under computer control. Selecting numbers 1 through 5 set the joy-stick sensitivity, and hence maximum motor velocity.
You may add locations by clicking on them in the Image window when the "+" image tool is selected, or by moving the micropipette to that location and clicking on the "+" tool. You may save the resulting list to the clipboard and you may move any pipette to any location in the list. This list is used for example, to find a set of locations during an assay phase of the experiment and later visit those sites during a data collection phase. Entries may be edited and keyboard short-cuts simplify operation. The clipboard copy is a tab-delimited text file. |
The Image window permits you to continuously observe your research specimen and move electrodes with point and click operations. Various tools permit calibrating objectives, camera and electrodes and setting the origin (tying the electrode tip to a screen location and level of focus). Graphics in the window show you the location of the point where the electrode tip was tied to the screen. The Image window is optional: Target can use the image from other image capture programs running in the background. This is done by positioning the mouse over the external image window, and typing a key. Here is Target's Image window:
The tools in the Image window are, from top to bottom: select, calibrate, set origin, target, target direct, incremental calibration, and add-to-list.
MacOSX note: Target supports only some cameras. For those cameras that are not supported, and for those situations where the image is supplied by another imaging program, the tools of the Image window are provided by an Image Tools window. Although a little awkward (except on laptop computers), mouse clicks are simulated by the space bar. The Windows version uses invisible windows and so the Image Tools window is not useful.
Contact Jim Nash for copies of software.
Third party communication via DLL
TargetDLL. for Windows XP - Third party communications via enclosed DLL or by compiling and linking to enclosed source file.
Known problems:
Setting of surface level assumes par-focality. This means that the surface level should be set under highest magnification and not changed, and the safe level offset (releative to surface level) should be larger than the error in par-focality.
Original design by Dr. Jeff Smith, Dr. Naohiro Koshiya, and Jim Nash.
Programming by Jim Nash and Dr. Larry Woltz.
Testing and ideas by Brian Howie, Dr. Hide Koizumi, Dr. Naoshira Koshiya,
Dr. Lorin Milescu, Brian Gorman
The List Window holds locations in a global coordinate
system defined to include image, focus, and stage. The coordinate system
is aligned with the image, with origin in the upper left corner of the image
when the "preferred" objective is selected, and when the stage
and focus locations are zero. Prior to use of the List Window, you select
the preferred objective, move to a field of view you choose and can easily
return to, focus on the surface of the tissue sample, and set the origin
of the stage and focus by clicking on the two origin setting buttons in
the List Window that have a little "o".