Table of Contents 1. Image processing Measurements System Tools...10

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1 Introduction

2 Table of Contents 1 An Overview of ScopeImage Advanced...2 Features:...2 Function introduction Image processing Image Import and Export Open image file Import image form TWAIN support devices Image Export Image Processing Geometrical Transform (Warping) Gray Value Image Processing Color Balance Contrast Hue/Saturation Measurements Image Measurements Points Measure Tool Line Measure Tool Circle Measure Tool Rectangle Measure Tool Angle Measure Tool Measure Tool s Color Setting Measure graph and Image Merge Measurement results Clear Measurement data Create Measurement graph Print Measurement Results System Tools ROI (Region of Interest) Tools Zoom Tools Paint Tools Sign Tools Other Tools

3 An Overview of ScopeImage Advanced ScopeImage Advanced is a 24bits real color microscopy image processing software based years of experience in the field of digital image processing and analysis, which provided a large number of image processing and measurement tools, suitable for all kinds of microscopy image processing and measurement tasks. High reliability and robust performance, easy to use, makes it a powerful tool to the user in scientific and research fields. Digital Video record, playback, snap, processing measurement Fig.1 The software interface Features: Live Video and static Image acquisition ScopeImage Advanced provides high-quality capture and playback of multimedia streams. It supports a wide variety of formats, including Advanced Streaming Format (ASF), Motion Picture Experts Group (MPEG), Audio-Video Interleaved (AVI), even MPEG Audio Layer-3 (MP3), and WAV files. It supports capture using Windows Driver Model (WDM) devices or older Video for Windows devices. TWAIN Tool The TWAIN Tool allows the import of TWAIN format image from digital cameras, scanners, and many other TWAIN-compatible devices, such as USB cameras. Video Preview Tool Video preview tool support capture image from the video device compatible with MS WMD and VFW. Video Capture Tools Video Capture tool allows you to capture real time video streams to the AVI files at different capture rate and time limit. User can use playback tool to view the video clips at later time. 2

4 Video Playback Tool The Playback tool allows you to play AVI, MPEG video clips, single-stepped, paused and allows you to select and save individual images for further image processing and measurement operation. Image Processing Tools Extensive possibilities of contrast enhancement, shade correction, color adjustment, edge detection, gray image morphological filters, digital image warping, image arithmetic operation is available for improved visual representation of images. ROI Tool Four kinds ROI (Region of interest) tools allow you to select Rectangle, Ellipse and arbitrary shaped region on the image. Sign Tools Sign tools allow you to add text, arrows and scale ruler signs on the images at arbitrary position. The size and color of the signs can be specified. Calibration Tool The calibration Tool calibrates the system to real-world coordinates and units of measure with sub-pixel accuracy. Calibration parameter data sets can be saved to a file for different camera and lens. Measurement Tools Measurement tool allows you to measure point position, distance between points, lines, area and center point of rectangle and circle, angles of edges etc. The measurement ruler and results data can be added to the image with specified color. Both measurement graph and image merged with the measurement graph can be printed. User can specify the measurement tolerance; PASS/FAIL detection can be performed with real time. Function introduction 1. Image processing 1.1 Image Import and Export Before image processing, we first explain how to import and export images. ScopeImage Advanced provides five methods to import image data to the software Open image file Click on the toolbar, the image can be loaded form an image file previously saved to disk Import image form TWAIN support devices N ote: Before perform capture, you must select scanner in the menu. Procedures are as follows. First click select scanner in the file menu (Fig 1), a dialog (Fig 2) will be displayed screen, select you current device and click select. on the 3

5 Fig 1 Fig Image Export Click button to print images on the printer. Click button to save an image to the disc. 1.2 Image Processing Image processing is used to improving visual representation of images Geometrical Transform (Warping) Geometrical Transform used to alter image shape and position. There are 5 Geometrical Transform tools,you select these tools on the warping toolbar. Horizontal flip Vertical flip, Flip the image horizontally, along the vertical axis., Flip the image vertically, along the horizontal axis. Image warping menu Zoom Transform,Change image size and position. When the mouse is on the control point, Press mouse left button and move the control point on the adjustment frame, you can change the size of the image. When the mouse is inside the adjustment fram e, keep pressing the left button; you can move the image in different directions. Image Rotate,Rotate tool is used to alter the image angle and position. Using mouse to rotate or move the adjustment frame, you can change the image angle and position. Free Transform,Free Transform used to change the image shape freely. Using mouse to move the control point on the adjustment frame, you can change the shape of the image freely. Net warping can warp the image.,using mouse to move the control point on the net, you Control frame Gray Value Image Processing a. Invert Inverts the image, converts positive image to negative image,or converts negative image to positive image. 4

6 b. Grayscale Delete color information of the image, and converts the image to grayscale image. c. Shade Correction Shade correction is used to correct systematic errors in the image brightness, usually caused by uneven illumination of the sample. There are three shade correction methods in the software. Use them to produce different results. d. Field Shade Correction First capture an image under the same illumination condition but without the sample and selects menu item reference image to save it as a reference image. Second capture the image need to correct, and select menu item shade correction, a dialog box will request an reference image, press load button, and slide the slider control button, will produce different correction results. e. Image Smooth ScopeImage Advanced offers the following smooth filters: Average, Median and Low Pass. Average filter smoothes an image by replacing the center pixel in a neighborhood with the average value of the neighborhood. Median filter smoothes an image by modifying pixels that varies significantly from their surroundings. This is accomplished by replacing the center pixel in a neighborhood with the median value of the neighborhood. This filter is particularly effective at removing random spots or points that vary significantly from the background and high-impulse noise from an image. Low Pass filter blurs an image by modifying a pixel value to be more like its neighbors. This eliminates harsh edges by reducing the intensity differences between adjacent pixels. All these filters can be used to blur an image to eliminate details or random image noise. f. Edge detection ScopeImage Advanced offers the following Edge detectors: Roberts, Sobel, Laplacian, Kirsch, Prewitt, Horizontal Edge, Vertical Edge, Upright edge, Down Left Edge. These edge detectors extract and enhance edges and contours in an image by expressing intensity differences between neighboring pixels as an intensity value. Roberts filter highlighted edges against a dark background; Sobel filter is less sensitive to image noise. This generally results in an image with smoother, more pronounced outlines of only the principal edges. The Roberts filter enhances all edges within an image, even those introduced by noise. It highlighted edges against a dark background, and can get good results for binary image. So if your image appears black following a Roberts s filter, use your brightness, contrast to lighten the image and bring out the edge detail. Sobel filter is less sensitive to image noise. This generally results in an image with smoother, more pronounced outlines of only the principal edges. Laplacian results are very similar to those of Hi-Pass. It produces an image with harsh intensity transitions, and results in an image with only edges of high contrast visible. Kirsch, and Prewitt can get thick edges; they work in the way similar to pattern matching process. 5

7 Horizontal Edge detects horizontal edges, so used it when horizontal features need to be extracted from an image. Vertical Edge extracts vertical edges in an image by highlighting pixels with significant intensity differences from those to the left and right of it. It produces an image with just its vertical edges visible against a flat background. Vertical Edge filtering is used when vertical features need to be extracted from an image. You can select suitable edge detectors for different images. g. Image Arithmetic Image Arithmetic performs logic operations and arithmetic upon the active image, or AOI. Such operations are performed between two images. When you select the arithmetic command, the Image calculation dialog box will be presented. Within this box, select the kind of calculation you want to perform, the input image and out put image you want it to work with. Output: The first image (which is the current active image) to use for the operation, this image will be replaced by the result image. Input: Select one of the files in this list box to tell ScopeImage Advanced what image to use for the operation as the second image. Channel: Tell ScopeImage Advanced which channel (Red, Green, Blue or all RGB channel), will join the calculation. Calculate: Select the kind of operation you want to perform from the drop-down list box. There are total 15 kinds of calculations as follows: Add: add the specified two images. Minus: subtract the specified input image from the active image. E.g. subtracting A smoothed image from an original image to emphasize high-contrast elements. Multiply: multiply the active image by the specified input image. Divide: divide the active image by the specified input image. AND: perform a logical "AND" between your active image and the specified input image. NOT: perform a logical "NOT" on the bit values of your active image and input image. XOR: perform a logical "XOR" (exclusive OR) between your active image and the specified input image. AND NOT: perform a logical "NAND" between your active image and the specified input image. OR AND: perform a logical "AND OR" between your active image and the specified input image. Diff: obtain the absolute value between the specified input image and the active image. Max: replace each pixel with the largest value of the two images. Min: replace each pixel with the smallest value of the two images. Feature and: perform feature and between the specified input image and the active image. Mask and: perform mask and between the specified input image and the active image Color Balance Color Balance tool is used to adjust the Red (Cyan), Green (Magenta) and Blue (Yellow) value of your images. C/R: Move this slider right to increase the image Red (decrease Cyan) value, or left to decrease 6

8 the image Red (increase Cyan) value. A value of 0 represents no change to the image values. M/G: Move this slider right to increase the image Green (decrease Magenta) value, or left to decrease the image Green (increase Magenta) value. A value of 0 represents no change to the image values. Y/B: Move this slider right to increase the image Blue (decrease Yellow) value, or left to decrease the image Blue (increase Yellow) value. A value of 0 represents no change to the image values Contrast Contrast adjustment tool is used to modify the visual qualities of your image. There are two slider controls for Brightness and Contrast characteristic. Brightness: Move this slider right to increase, or left to decrease the image brightness. A value of 0 represents no change to the image values. Contrast: Move this slider right to increase, or left to decrease, the image contrast. A value of 0 represents no change to the image values Hue/Saturation Hue/Saturation tool is used to adjust the Hue, Saturation and brightness value of your images. Hue: Move this slider right to increase or left to decrease the image Hue value. A value of 0 represents no change to the image values. Saturation: Move this slider right to increase or left to decrease the image Saturation value. A value of 0 represents no change to the image values. Brightness: Move this slider right to increase or left to decrease the image Brightness value. A value of 0 represents no change to the image values. Hue is the color reflected from or transmitted through an object. In common use, hue is identified by the name of the color such as red, orange, or green. Saturation, sometimes called chroma, is the strength or purity of the color. Saturation represents the amount of gray in proportion to the hue, measured as a percentage from 0% (gray) to 100% (fully saturated). Brightness is the relative lightness or darkness of the color, usually measured as a percentage from 0% (black) to 00% (white). 2. Measurements 2.1 Image Measurements Using the Measurements tools in the Measurement toolbar (or measurement menu), you can measure the length of lines, the area of circles, and the angles of edges that you define manually.. The Measurements tools also let you measure distances between any two points, and perform tolerance testing. To measure object on the image, you have to create measurement frames or features on the image first. The measurement frame is consisting of control points; coordinate points, distance lines and texts. You adjust the measurement frames using the mouse manually. 7

9 2.1.1 Points Measure Tool Button is used to create a point measurement feature on the current image. Button is used to select a point. You can move the point feature on the current image when press mouse left button. You can delete that point using Delete key on the keyboard when the point is selected. Button is used to create a line connecting two points, which gives the distance between the tow points Line Measure Tool Line measure tool is used to measure the parameter of the line between two points. Button creates a line measurement feature on the image. Button is used to select a line measurement frame. You can move the line feature on the current image when press mouse left button and move the mouse cursor when the mouse hits the line. You can change the line s position and length by adjusting the end the line s end points. You can delete that line using Delete key on the keyboard when the line is selected Circle Measure Tool is used to create a circle measurement frame on the current image, which is used to measure circle parameters, such as diameter, perimeter, area, and center position. is used to create a circle measurement feature on the current image by using three points. is used to set active circle and adjust circle size and position. You can move the circle on the current image when press mouse left button at the circle center point. You can change the size of the circle by moving the adjust point on the circle edge. And you can delete that circle by using Delete key on the keyboard when the circle is selected Rectangle Measure Tool is used to create a rectangle measurement feature on the current image, which is used to measure rectangle parameters, such as perimeter, area, and center position. is used to set active rectangle and adjust rectangle size and position. You can move the rectangle on the current image when press mouse left button inside the rectangle. You can change the size of the rectangle by moving the adjust point at the rectangle corners. You can delete that rectangle by using Delete key on the keyboard when the rectangle is selected. 8

10 2.1.5 Angle Measure Tool is used to create an angle measurement feature on the image. There are three angles are measured, the first angle is the angle between the first edge and X axis. The second angle is the angle between the second edge and X axis. The third angle is the angle between the two edges. The positions of the edge end points are also measured. is used to set active angle measurement feature and adjust its position. You can move the angle measurement feature on the image when press mouse left button at the edge cross point. You can change the angle of the angle feature by moving the two end points. And you can delete that ang le feature by using Delete key on the keyboard when the angle feature is selected Measure Tool s Color Setting Every measurement frame is consisting of control points; coordinate points, distance lines and text. You can modify the color of them by press the button. When active that button, a Measure Tool s Color Setting dialog box will be displayed. You can select the color you wanted Measure graph and Image Merge This command merges the measurement frames with the current image. You can save or print the merged image. 2.2 Measurement results All the measured data are displayed in the measurement results dialog box. You can save, print, delete or clear the measurement data using that dialog box. Check tolerance test check box, ScopeImage Advanced will perform pass/fail tolerances test. Select a data item in the dialog box and press delete key, you can delete the data selected. Click output button, the output data dialog box will be displayed. There are nearly 40 measurement data items can be selected. Only the selected data items will be print out or saved. The Add button is used to insert measurements of the features to be output. The Delete button is used to delete a selected data item. Add all button select all data items listed in the right list box. Clear button clear all items selected in the list box. You can save, load and reset the selected items if needed. Click tolerance button, the tolerance set dialog box will be displayed. You can set the upper limits and lower limits of the tolerances. ScopeImage Advanced will make tolerance testing using these data. For example, if the standard diameter of the circle is 100mm, and the upper limit is 0.5mm, the lower limit is 0.5. All circles with diameter between 99.5 and will pass the test. You can save the tolerance data for later use. 2.3 Clear Measurement data Clear all measured data. 9

11 2.4 Create Measurement graph Create a white image only with measurement data and graph. 2.5 Print Measurement Results Print measured image. You can add text and adjust the size and position of the image to be printed. Image measurement 3. System Tools ScopeImage Advanced offers more than 50 tools to assist image processing and measurements. 3.1 ROI (Region of Interest) Tools There are four ROI tools in the toolbox The lasso tool let you draw freehand segments of a ROI border. To use the lasso tool: a. Select the lasso tool in the toolbox. b. Drag to draw a freehand ROI border. c. Double click to end the selection. Hold down the Ctrl key while using lasso tool, you will erase the selected ROI The ellipse tool is used to select ellipse ROI To use the ellipse tool: a. Select the ellipse tool in the toolbox. b. Drag to draw an ellipse ROI border. c. Release mouse button to end the selection. Hold down the Ctrl key while using ellipse tool, you will erase the selected ROI. 10

12 3.1.3 The rectangle tool let you draw rectangle ROI border. To use the rectangle tool: a. Select the rectangle tool in the toolbox. b. Drag to draw a rectangle ROI border. c. Release mouse button to end the selection. Hold down the Ctrl key while using rectangle tool, you can erase the selected ROI The ROI cut tool let you to delete the selected ROI. To use the ROI cut tool: a. Select the ROI cut tool in the toolbox. b. Click the ROI you want to delete, the region will be deleted. 3.2 Zoom Tools ScopeImage Advanced offers two zoom tools The Zoom Tool let you to magnify the image. To use the zoom tool: a. Select the zoom tool. The pointer becomes a magnifying glass with a plus sign in its center. b. Click the area you want to magnify. Each click magnifies the image to the next preset percentage, centering the display on the point you click. When the image has reached its maximum magnification level of 1600%, the magnifying glass appears empty. Hold down the Ctrl key while using zoom tool; you will zoom out the image. The pointer becomes a magnifying glass with a minus sign in its center. When the image has reached its minimum magnification level of 1.0%, the magnifying glass appears empty The Magnifying Glass let you to view the image in the way as using a magnifying glass. To use the magnifying glass tool: a. Select the magnifying glass tool in the toolbox. b. Move the glass to view the magnified image. c. Double click to exit magnifying glass tool. 3.3 Paint Tools The line tool is used to draw straight lines. To use the line tool: a. Select the line tool in the toolbox. b. Position the cross pointer where you want the straight line to begin, Hold the button and drag to draw the line. c. Release the button to end line drawing. You can choose the pen size and color using the sign setting tool or eyedropper tool The Fill Tool let you to fill an image region with specified color. 11

13 To use the fill tool: a. Select the fill tool in the toolbox. b. Position the paint bucket pointer where you want to fill and click. Note1: You can choose the color using the sign setting tool or eyedropper tool. Note2: You can choose the fill tolerance using the sign-setting tool. Choose a low tolerance val ue to fill a region with colors very similar to the pixel you click, or a higher value to fill a region with broader range of colors The Eyedropper Tool let you to pick colors on the image. To use the eyedropper tool: a. Select the ey edropper tool in the toolbox. b. Position the eyedropper pointer where the color you want to pick and click The Pencil Tool let you to paint on the image like using a pencil. To use the pencil tool: a. Select the pencil tool in the toolbox. b. Position the pencil pointer where you want to begin to paint. Hold the button and drag to draw with the pencil. c. Release the button to end drawing. You can choose the pencil size and color using the sign setting tool or eyedropper tool. You can also use up and down key (, ) to increase or decrease the pencil size while using the pencil tool The Ellipse Paint Tool let you to draw an ellipse on the image. To use the ellipse tool: a. Select the ellips e paint tool in the toolbox. b. Drag to draw an ellipse on the image. c. Release mouse button to end the draw. You can choose the pen size and color using the sign setting tool or eyedropper tool The Filled Ellipse Paint Tool let you to draw a filled ellipse o n the image. To use the filled ellipse tool: a. Select the filled ellipse paint tool in the toolbox. b. Drag to draw a filled ellipse on the image. c. Release mouse button to end the draw. You can choose the pen size and color using the sign setting tool or eyedropper tool The Rectangle Paint Tool let you to draw a rectangle on the image. To use the rectangle tool: a. Select the rectangle paint tool in the toolbox. b. Drag to draw a rectangle on the image. c. Release mouse button to end the draw. You can choose the pen size and color using the sign setting tool or eyedropper tool. 12

14 3.3.8 The Filled Rectangle Paint Tool let you to draw a rectangle on the image. To use the filled rectangle tool: a. Select the filled rectangle paint tool in the toolbox. b. Drag to draw a filled rectangle on the image. c. Release mouse button to end the draw. You can choose the pen size and color using the sign setting tool or eyedropper tool The Polygon Paint Tool is used to paint a polygon on the image. To use the polygon paint tool: a. Select the polygon paint tool in the toolbox. b. Click in the image to set the starting point. c. Position the pointer where you want the first straight segment to end, and click. Continue clicking to set endpoints for subsequent segments. d. Double-click the polygonal lasso tool pointer to close the polygon. You can choose the pen size and color using the sign setting tool or eyedropper tool The Filled Polygon Paint Tool is used to paint a filled polygon on the image. To use the filled polygon paint tool: a. Select the filled polygon paint tool in the toolbox. b. Click in the image to set the starting point. c. Position the pointer where you want the first straight segment to end, and click. Continue clicking to set endpoints for subsequent segments. d. Double-click the polygonal lasso tool pointer to close the polygon. You can choose the pen size and color using the sign setting tool or eyedropper tool. 3.4 Sign Tools Sc ale Tool The Scale Sign Tool let you to paint a scale sign on the image. The scale presents the real size of the image. To use the scale sign tool: a. Select the scale sign tool in the toolbox. The pointer becomes a cross. b. Move the cross cursor to the desired position on the image and press the left button, a scale rule will be painted on the image. c. Adjust the position of the rule when mouse becomes a hand. Adjust the size of the rule when mouse position is at the end points of the ruler, the cursor becomes a double arrow, and you can modify the size of the ruler at this time. You must calibrate the software before using the scale sign tool. You can change the ruler color using the sign-setting tool Arrow Tool The arrow Sign Tool is used to add arrow signs on the image. To use the scale sign tool: a. Select the arrow sign tool in the toolbox. The pointer becomes a cross. 13

15 b. Move the cross cursor to desired position on the image and press the left button, a arrow will painted on the image. c. Press up and down arrow key to adjust the size of the arrow. Press left and right key, to rotate the arrow. You can change the arrow s color using the sign-setting tool Text Tool The Text Tool let you to add text on the image. To use the text tool: a. Select the text tool in the toolbox. The pointer becomes a (T). b. Move the T cursor to the desired position on the image and click, the text edit dialog box will be displayed. c. Enter text as desired in the dialog box s edit box. Press Font button to select the font and color of the text. Press OK to paint the text on the image. Note1: Hold a nd drag the mouse, you can move the text painted on the image. Note2: Double-click the text; you can modify the text painted on the image Sign Setting Tool The Sign Setting Tool let you to set the pain tools color, size and tolerance (for fill tool). To use the Sign Setting tool: a. Select the Sign Setting tool in the toolbox. The sign setting dialog box will be displayed. b. Click on the palette to select the color desired. Adjust the slider in the slide control to modify tool size and tolerance. c. Press OK to close the dialog box. You can use eyedropper to select colors on the image. 3.5 Other Tools Undo Tool The undo tool let you undo the last action Redo Tool The redo tool let you redo the previoously undone action Crop Tool The crop tool let you to cut off the image not included in the ROIs Cut Tool The cut Tool let y ou to cut the image and transfer the image to the clipboard. If no ROI exists on the image, the whole image will be processed Copy Tool The copy Tool is used to copy the selected image to the clipboard. If there is no ROIs exist on the 14

16 image, the whole image will be processed Paste Tool The paste Tool let you paste an image on the clipboard to the current active image. If there is a ROI on the image, the image will paste into the ROI Color Balance The color Balance tool is used to adjust the Red (Cyan), Green (Magenta) and Blue (Yellow) value of your images. C/R: Move this slider right to increase the image Red (decrease Cyan) value, or left to decrease the image Red (increase Cyan) value. A value of 0 represents no change to the image values. M/G: Move this slider right to increase the image Green (decrease Magenta) value, or left to decrease the image Green (increase Magenta) value. A value of 0 represents no change to the image values. Y/B: Move this slider right to increase the image Blue (decrease Yellow) value, or left to decrease the image Blue (increase Yellow) value. A value of 0 represents no change to the image values Contrast Contrast adjustment tool is used to modify the visual qualities of your image. There are two slider controls for Brightness and Contrast characteristic. Brightness: Move this slider right to increase, or left to decrease the image brightness. A value of 0 represents no change to the image values. Contrast: Move this slider right to increase, or left to decrease the image contrast. A value of 0 represents no change to the image values Hue/Saturation The Hue/Saturation tool is used to adjust the Hue, Saturation and brightness value of your images. Hue: Move this slider right to increase or left to decrease the image Hue value. A value of 0 represents no change to the image values. Saturation: Move this slider right to increase or left to decrease the image Saturation value. A value of 0 represents no change to the image values. Brightness: Move this slider right to increase or left to decrease the image Brightness value. A value of 0 represents no change to the image values. Hue is the color reflected from or transmitted through an object. In common use, hue is identified by the name of the color such as red, orange, or green. Saturation, sometimes called chroma, is the strength or purity of the color. Saturation represents the amount of gray in proportion to the hue, measured as a percentage from 0% (gray) to 100% (fully saturated). Brightness is the relative lightness or darkness of the color, usually measured as a percentage from 0% (black) to 00% (white) Calibrating Tool The calibration Tool calibrates the system to real-world coordinates and units of measure with 15

17 sub-pixel accuracy. Calibration parameter data sets can be saved to a file for different camera and lens. To use the calibrating tool: a. Put a micro scale horizontally in the filed of view, and capture an image of the scale. b. Select the calibrating tool in the toolbar, A cross ruler and a calibration dialog box are displayed. c. Select the scale s unit using the list box in the dialog box. d. Enter the scale length you want to measure in the X ruler size edit box. e. Move and adjust the ruler cursor s horizontal arm to match the scale marker on the ruler image. f. Press OK. g. Do the same for the vertical direction. Note1: You should calibrate the software before doing measurement works. Note2: You should calibrate the both in horizontal and vertical direction. Note3: You can save and load the calibration data by pressing save and load button on the calibrating dialog box. Calibration dialog box 16