SPOT 4.1 Basic and Advanced Software Manual for SPOT Insight ...

Ch. 3 – Basic Program Mode• There are six view/exposure options at the bottom of the Live Image window:RestartPause/ResumeCloseFit to WindowReset AreaFaster ReadoutResample the brightness of the image, compute new exposure times, and restart the live imagedisplay. This is useful for updating exposure times when specimen brightness has changed.Freeze the live image window. This is useful when you are changing a specimen and want topreserve the last view. In addition, to use the Transfer option (see below), you must first Pausethe live image view. Click on Resume to restart the live image viewClose the Live Image window.Size images to automatically fit the resizable display window.Return to the non-zoomed view of the live image.Faster Readout enables you to “speed up” the live image display (higher frame rates) in caseswhere you are willing to sacrifice some resolution for a higher frame rate.Computing White Balance Values (Insight Color Mosaic & 3Shot)White balance is the ratio of red, green, and blue gains necessary to achieve the proper colorrendition for an image. Many factors impact the coloration of an image capture, includinglamp voltage, the coloration of the glass used in the slide, and the coloration of the glass used in the lensesof the microscope’s objectives. To minimize the impact of these factors, use the Compute White Balancefeature. When you perform a white balance calculation, SPOT samples the color of the light from yourlight source after it has passed through the slide and the objective, and then calculates the red, green, andblue gains needed to produce images where the white areas are white and the color areas are true.You should perform a new white balance calculation when:• Starting a new image capture session.• Changing the lamp voltage.• Switching to a different objective on the microscope.To compute new white balance values, follow these steps:1. Set the illumination and the objective that you will use for the image capture.2. If you have not done so, select your Image Type (see above). Each Image Type has its own set ofwhite balance values. Any white balance values that you compute for an image type remain linked tothat image type until you compute new values. When selecting a new image type, remember that youwill be using the white balance values linked to that image type.3. Click on the White Balance button.4. Show the camera a sample of white light. This step varies according to microscopy technique andImage Type (sample), as shown in the table below:Image TypeBrightfieldtransmittedlightDarkfieldtransmittedlightFluorescenceNomarskiDIC/HoffmanPhase contrastWhite Balance TechniquePosition the slide so that the specimen is not in the field of view, but light is going through the slidenear the specimen.Perform the white balance using the brightfield mode on your microscope. See Brightfield–transmitted light above.Use the Match Color featureFor Nomarski DIC: Adjust the Nomarski prism until the image is in either the “brightfield” or the“gray scale” view.For Hoffman: Perform the white balance directly on the Hoffman image.Position the slide so that the specimen is not in the field of view, but light is going through the slidenear the specimen.User Guide to the SPOT Insight Camera 43