Center for nanoscale science and engineering Standard Operating Procedure



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Center for nanoscale science and engineering

Standard Operating Procedure

Microscope Software









Brian Wajdyk








Important

  • Images are not to be saved to the computer. They will be deleted without warning. Use a USB drive or burn a CD.

  • No additional software is too installed on the machine without prior approval by Dr. May or Brian Wajdyk.

  • The computer is on the network/internet. Please be careful of which sites you visit.

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Image Aqusition



Figure
Measurement



Figure
Operating Procedure

  1. Preparation

    1. Find the area of the sample to be imaged in the normal manner. Center the area of interest as much as possible. The field of view is less wide for the camera than seen through the eyepieces.

  2. Load the software

    1. Logon to the computer if necessary. Logon ID: Microscope User. There is no password.

    2. Double click the AxioVision icon on the desktop.

  3. Select Camera

    1. There is no instance that you should have to change the camera source. However if you get no image you can check to troubleshoot.

    2. To change or check the camera go to the File menu and scroll down to Select TWAIN device.

    3. The device is that should be selected is called “UCMOS10000PA 1.2 (32-32)”

  4. Start image preview

    1. To start the image acquisition go to the File menu again this time select TWAIN-camera image acquisition.

  5. Acquire image

    1. A live image will appear on the screen. It will be slightly out of focus compared to what you saw through the eyepieces. We may make it parfocal in the future.

    2. Focus while watching the live image. Be VERY careful to not touch your sample to the lens.

    3. The advised Preview resolution is 912*684 because the refresh rate is high enough for near instant feedback as you change the focus and/or positioning of the sample.

    4. You may change to resolution of the captured image to whatever you wish by using the pull down menu labeled still. Full resolution is 3664*2748 (10MP).

    5. When the image is acceptable click on the capture button. The larger the image the longer the processing time. At 3664*2748 it takes about 30sec.

    6. Eventually the acquisition window closes and you will see your image in the main window.

  6. Saving the image

    1. To save the image you can go back to the File menu and select save as or select the 3rd icon on the top toolbar.

    2. You can save the image in one of four formats: a proprietary format for the software, JPEG, BMP, or TIFF.

    3. No images are to be saved to the hard drive. Images saved to the hard drive will be deleted without notification.

  7. Image Manipulation

    1. By default, the image is shown at full resolution. This makes it appeared to be zoomed in at a higher magnification.

    2. You will need to zoom out in order to see the entire image. This can be done by selecting the magnifying glass under the image with the vertical arrows (3rd icon from the left). You can select anything in between by using the magnifying glass with the + to increase the zoom or alternatively the magnifying glass with the – to decrease it. These manipulations do not change the actual image in anyway. Only the way viewed is changed.

  8. Measurements

    1. Measurements are very accurate using the software. To measure a feature on a image go to the Workarea in the upper left section of the screen and click on Scalings.

    2. The available scalings will be shown. Default is Pixels so you will need to select the scaling based on the microscope objective used and the measurement unit desired. For example to measure a feature you obtained a image with the 10x objective in millimeters, you would click on the scaling marked “10x mm”.

    3. Once you have selected the scaling you can hit the “Activate” button. All further measurements will be made using this scaling until the software is shut down.

    4. Now you have to apply that scaling to your current image by hitting the “apply to Image” button.

    5. Now you are able to measure your features. Select the Measure pull down menu to Length. Note: You may want to zoom in on your area of interest if a more accurate measurement is needed. The software won’t be more accurate because it counts pixels, but you might have an easier time selecting the correct pixels at full resolution depending on your sample.

    6. Click and drag a line then click again to define the area to be measured. The result will show up on the screen.

    7. Often you will want to add a Scale bar to your image. This is because the true magnification varies based on screen resolution, display resolution, and scaling when printing. The Scale bar will always be correct relative to the features no matter how the image is displayed. To add the scale bar press the scale bar icon in the top toolbar.

    8. Click and drag as before.

  9. More advanced features

    1. The software is capable of much more. For instance you may want to change the color of the text. This is beyond the scope of this operating procedure. However the full manual is available on the computer desktop. Feel free to make a copy and take it with you.


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