Motic Images Plus 3.0 User Manual

Motic Images Plus 3.0 ML Software

Windows OS User Manual

Introduction 05
Menus and tools 05
File 06

New 06
Open 07
Save 07
Save as 07
Export 07
Capture 08
Image Comparison 08
Album 09
Folder 09
Amalgamation 10
Report 11
Report tool bar 12
New Page 12
Load 12
Save 12
Print Current Page 13
Print 13
Edit 13
Text 13
Color 14
Font 14
Settings 14
Capture 14
Audio 15
Exit 15

Edit 15

Rectangle Marquee 16
Circle Marquee 16
Ellipse Marquee 16
Polygon Marquee 16
Irregular Marquee 16

CONTENTS (Linked)

Motic Images Plus 3.0 ML Software | Windows OS User Manual

Image 17

Brightness/Contrast 17
Hue/Saturation 18
RGB 18
Mirror 18

Vertical 19

Rotate 90° 19
Gray Scale 19
Invert 19
Image Size 19
Filter 19
User defined Filter 20

Measure 21

Line 21
Rectangle 21
Circle 21
Circle (3 points) 21
Arc 22
Ellipse 22
Polygon 22
Freehand 22
Freehand Line 22
Angle 22
Parallel 22
Marker 22
Auto Calculation 23
Calibration Wizard 24
Calibration Table 25

Language 25
Style 25
View 25
Help 26

About 26
Help 26

Control Panel 26

Edit 27
Text 27
Paint 27
Audio 28
Measure 30
Magnifier 32

Preview Window 33
How to use the Popup Menus... 33

...in the Image window 33
...in the Album window 35

Calibration... 36

...with Calibration Circle 36

...with Scale Cross 37

...with Scale Line 38

Motic Images Plus 3.0 ML is multi-language software offered by Motic China Group Co., Ltd. It is
especially designed to adjust interactively a live image supplied by a Motic USB camera, capture the
optimized image and perform further analyzing and measuring tasks.

Opening Motic Images Plus 3.0 ML will display the following interface:

The interface is composed of the Title Bar, Menu Bar, Tool Bar, Image Window, Control Panel and

Preview Window.

Menus and tools

The Menu Bar includes the functions File, Edit, Image, Measure, Language, Style, View and Help.
Click the buttons on the Menu Bar to display the corresponding tools.

Menus and tools / File

File includes the following commands: New, Open,Save, Save As, Export, Capture, Preview, Image
Comparison, Album, Report, Settings and Exit. Shown below is the corresponding tool bar:

Menus and tools / File / New

The New command enables to create a new image file:

Select the format of the image file (jpg; bmp; tiff; png) and the file name. “png” is a proprietary Motic
format which can only be opened with Motic Images Software. Define the image size. Capture Folder
defines the location where the new image file should be saved.

Menus and tools / File / Open

The Open command allows users to open already existing image files:

Menus and tools / File / Save

The Save command allows to save the current image in the selected format.

Menus and tools / File / Save As

The Save As command provides another method for saving the current image. The following dialog box
will display allowing users to save the image to a different destination and in a different format than its
source.

Menus and tools / File / Export

Export saves the current image in a selected path.

Menus and tools / File / Capture

Selecting the Capture command will bring up the Motic Live Imaging Module to handle images seen
through the microscope connected to the computer.

Note: The capture device can be preset using the Setting command of the File menu or the Setting
button on the File Toolbar:

Menus and tools / File / Image Comparison

The Image Comparison command opens a window composed of two frames for a side-by-side
comparison of two images.

There are four ways to load the images for comparison:

1. Double click on either frame to load images from the local disk.

2. Right click within either frame and select Load Image from the shortcut menu to load the desired

images for comparison.

3. Images may also be dragged into either of the frames directly from the Album or Preview window.

4. Click “Load Image” button to import the selected image.

Menus and tools / File / Album

The Album command allows to open already existing images from their thumbnails. It also opens the
general WINDOWS explorer. Click the Album button to bring up the following window:

Menus and tools / File / Album / Folder

The upper column displays the Motic Images Plus 3.0 ML image folders, the images captured will be
saved here. The contents of an Image Folders on the left will be shown in a thumbnail display on the
right. Double click on a thumbnail selects a file for single image display; “Drag and Drop” from thumbnail
to Preview window is also possible. With Set default capture folder a predefined path for image storage
can be set.

Menus and tools / File / Album / Amalgamation

Amalgamation is a helpful tool for image overlay, especially for multiple stained samples in fluorescence.
Click the Amalgamation tab to display the following interface:

Select the desired images and drag them into the Amalgamation panel. Click the Amalgamation button
and a popup menu listing various amalgamation calculations will be displayed.

Select an amalgamation method and a dialog box will display to confirm the name of the resulting image.
Check the different options to find the best possible amalgamation result. The overlay will be saved in the
format “combination_1.”
Clicking the Remove/Remove All buttons will remove selected image.
Users can also remove an image in the Amalgamation panel by left click for selecting it; then clicking the
right mouse button to bring up the following popup menu:

Menus and tools / File / Report

The Report function is an integral part of Motic Images Plus 3.0 Software. Users are able to load images
and add text fields for printing or export purposes. A report will be saved under proprietary format
moticreport.mwp.

To close the report button click located in the top right corner.
Click the right mouse button in the Motic Report window to bring up the following popup menu:

Menus and tools / File / Report / Report Tool Bar

The Report Tool Bar is located at the top of the Report window and displays as follows:

Menus and tools / File / Report / Report tool Bar / New Page

The New Page button creates a new report page.

Menus and tools / File / Report / Report tool Bar / Load

The Load button brings up the following dialog box from which an existing report may be opened.

Menus and tools / File / Report / Report tool Bar / Save

The Save button brings up the Save As dialog box which allows users to save the report documents as
*.mwp format files.

Menus and tools / File / Report / Report tool Bar / Print Current Page

The Print Current Page button enables users to preview the current report.

Menus and tools / File / Report / Report tool Bar / Print Setting

The Print Setting button brings up the Print dialog box.

Menus and tools / File / Report / Edit / Text

Click the Text button and move the cursor to the location on the report where the text is to be displayed, click the left mouse button and begin typing.

Menus and tools / File / Report / Edit / Text / Color

The Color button brings up the Color Palette through which users are able to select a text color.

Menus and tools / File / Report / Edit / Text / Font

The Font button brings up the Font dialog box.

Menus and tools / File / Settings

The Settings command provides access to the parameter of Still Image capture, image sequences and
Audio options.

Menus and tools / File / Settings / Capture

Click the Capture tab to display the capture settings. Here, define the file name, image size, the
frequency and the maximum number of images to be captured automatically. To choose the Using
current date and time as file names option, click the respective check box.
Click OK to confirm.

Menus and tools / File / Settings / Audio

Click the check box to the left of the Auto Play option and the music selected in the Playlist of the Control
Panel will play automatically each time Motic Images Plus 3.0 ML is opened. Click OK to confirm.

Menus and tools / File / Exit

The Exit command closes the program after prompting users to save any unsaved files.

Menus and tools / Edit

The Edit Menu allows to define selected areas for further handling. The corresponding tool bar functions
are equivalent:

Rectangle Marquee

A Rectangle Marquee will be defined by clicking and dragging the left mouse button.
To move the selected rectangular region, place the mouse pointer on the side of the figure, and then click
and drag when the cross pointer appears.To change the size of the selected region, place the mouse
pointer on a corner of the rectangle, and then click and drag the mouse when the hand pointer appears.

Circle Marquee

The Circle Marquee will be defined by clicking the left mouse button to define the center and to drag for
size variation.To move the circle, place the mouse pointer at the center, then click and drag the mouse
when the cross pointer appears. To change the size of the selected region, place the mouse pointer at
the extreme point of the radius, then click and drag the mouse when the hand pointer appears.

Ellipse Marquee

The Ellipse Marquee is defined like a circle. To move the selected elliptical region, place the mouse
pointer at the center of the ellipse, then click and drag the mouse when the cross pointer appears.
To change the size of the selected region, place the mouse pointer at the extreme point of the major or
minor half axis, then click and drag the mouse when the hand pointer appears.

Polygon Marquee

A Polygon Marquee is defined by clicking and dragging the left mouse button.To move the polygon,
place the mouse pointer on the side of the polygon, then click and drag the mouse when the cross pointer
appears. To change the size of the selected region, place the mouse pointer on a corner of the polygon,
then click and drag the mouse when the hand pointer appears.

Irregular Marquee

With the Irregular Marquee command, users are able to define an irregular region by clicking and
dragging the mouse. The selected irregular region may be moved using the drag-and-drop method when
the cross pointer appears.

All selected areas can be deleted by or right mouse button: Deselect region

Menus and tools / Image

The Image Menu includes the all commands for a post-capture image correction. The corresponding tool
bar functions are shown below:

The RGB box displays the color information at the point of the cursor's current location.

The XY box displays the XY coordinates of the cursor's current location.

Menus and tools / Image / Brightness/Contrast

The Brightness/Contrast command displays separate sliders for both parameters.

The Default values are both set to “0”. For brightness, the value range is -255 to +255; for contrast it is

-127 to +127.

Menus and tools / Image / Hue/Saturation

Select the Hue/Saturation command to bring up the following window:

Hue values can be varied from -360 to +360; the default value is “0”. Saturation range is from -100 to
+100; default value is “0”.

Menus and tools / Image / RGB

Select the RGB command to bring up the following window:

The three sliders are used to adjust the red, green and blue color channels of the active image. The value
range always is from -100 to +100 with default value “0”.

Menus and tools / Image / Mirror

The Mirror command mirrors the current image horizontally.

Menus and tools / Image / Vertical

The Vertical command mirrors the current image vertically.

Menus and tools / Image / Rotate 90

The Rotate 90 command rotates the current image clockwise by 90 degrees.

Menus and tools / Image / Gray Scale

The Gray Scale command converts the current image into a B/W image.

Menus and tools / Image / Invert

The Invert command inverts the current image colors.

Menus and tools / Image / Image Size

Image Size brings up the Image Size dialog box. The units may be "Pixel" or "Percent".
Select the Constrain Proportions checkbox to keep the image proportions even though the image size
is changed. If Width or Height values are changed manually, the corresponding value will automatically
be adjusted to keep the image proportions.

Menus and tools / Image / Filter

The Filter command allows to apply several filters on the saved image.

Menus and tools / Image / User defined Filter

The User Define Filter command allows the individual configuration of a filter.

To define a new filter, click "New" and enter the name of the new filter. Enter values into the blanks to
create a filter matrix with divisor and bias.
To open an existing filter, select an item from the list and click "Apply".

Menus and tools / Measure

The Measure Menu includes the commands for measuring several geometrical figures as well as the
quantification tools Auto Segment, Auto Calculation, Calibration Wizard and Calibration Table.

Line

Line measures distances, diameter etc. Click the left mouse button and drag a line. The result is

displayed immediately.
Place the mouse pointer at the extreme point of the line (arrow → hand pointer) and change the length
interactively. Place the mouse pointer on the line, then click and drag the mouse when the cross pointer
appears to move the line.

Rectangle

The Rectangle command measures the width, height, area and perimeter of rectangles. Click and drag
the left mouse button on the image to create a rectangle.
Place the mouse pointer at the vertex of the rectangle, then click and drag the mouse when the hand
pointer appears to change the size of the rectangle. Place the pointer on an edge of the rectangle, then
click and drag the mouse when the cross pointer appears to move the rectangle.

Circle

The Circle command measures the radius, area and perimeter of circles.
Place the mouse pointer at the extreme point of the radius, then click and drag the mouse when the hand
pointer appears to change the size of the circle. Place the mouse pointer on the center of the circle, then
click and drag the mouse when the cross pointer appears to move the circle.

Circle (3 points)

The Circle (3 points) command measures the radius, area and perimeter of circles. Define a circle by 3
points. Place the mouse pointer on any of the three points on the circle, then click and drag the mouse
when the hand pointer appears to change the size of the circle. Place the mouse pointer on the circle,
then click and drag the mouse when the cross pointer appears to move the circle.

Arc

The Arc command allows users to measure the radius, perimeter, angle and arc length.
Click and drag the mouse to create an arc with 3 points. Place the mouse pointer at one red point of the
arc, then click and drag the mouse (arrow → hand pointer) to change the shape of the arc. Place the
mouse pointer on the arc, then click and drag the mouse (arrow → crosspointer) to move the arc.

Ellipse

The Ellipse command measures the major half axis, minor half axis, area and perimeter.
Place the mouse pointer on the extreme point of the major or minor half axis, then click and drag the
mouse when the hand pointer appears to change the size and shape of the ellipse. Place the mouse
pointer on the center of the ellipse, then click and drag the mouse when the cross pointer appears to
move the ellipse.

Polygon

The Polygon command measures the area and perimeter.
To complete a polygon, double click the left mouse button on the end point.
Place the mouse pointer on any edge of the polygon, then click and drag the mouse when the hand
pointer appears to change the size of the polygon. Place the mouse pointer on any point of the polygon,
then click and drag the mouse when the cross pointer appears to move the polygon.

Irregular

The Irregular command measures the area and perimeter of irregular figures. Click and drag the left
mouse button on the image to drawn an irregular image, release the button to complete the shape
automatically. Click and drag the mouse when the cross pointer appears to move the irregular figure.

Freehand line

This Freehand Line command allows users to measure the length of free lines.
Place the mouse pointer on the free line, then click and drag the mouse when the cross pointer appears
to move the free line.

Angle

3 points set by the left mouse button will define an Angle. Depending on the drawing, the internal or
external angle is measure. Place the mouse pointer at the end of the angle leg, then click and drag the
mouse (arrow → hand pointer) to change the angle. Place the mouse pointer on any point of the angle,
then click and drag the mouse (arrow → crosspointer) to move the angle.

Parallel

Parallel allows to measure the distance between 2 parallel lines. Draw the first line and end it by mouse

click. Moving the mouse creates a parallel line. Both lines can be moved by clicking the grey middle
marking (arrow →cross).

Marker

Using Marker allows to highlight defined points of interest. The x/y coordinates are displayed.

Auto Calculation

The Auto Calculation command provides detailed data of the objects segmented from the image.
The Auto Calculation Result window will then be displayed, such as number of objects, total area,
percentage of total area, minimum object area, maximum object area, average object area, minimum
object perimeter, maximum object perimeter, average object perimeter and the serial number, area and
perimeter of respective objects.
To access the data of a certain object, click on the object in the image window and the data for that object
will be highlighted.
Click on the data of the desired object(s) and the corresponding object(s) will flicker in the image window.
Click and drag on a particular data item to change its location in the list of results.

Click to bring up the Save As dialogue for export the calculation results as a Text file (*.txt)
or CVS file (*.cvs).

Calibration Wizard

A correct calibration is essential for any kind of measurement, for correct scale bars and grid dimensions.
The Calibration with Calibration Circle is based on the dot patterns metallized on the glass calibration
slide provided with Motic cameras. The calibration with Scale Cross and Scale Line are intended to be
used in case the smallest dot on the calibration slide is even too large to be captured completely by an
image.
Select the Calibration Wizard command to bring up the following interface:

Calibration with Calibration Circle

To calibrate with a calibration circle, select the respective Calibration Wizard tab. Click Load Image to
select a calibration circle from your image database. Confirm the objective lens used to capture the
selected image and input the diameter of the calibration circle. Click the Calibration button to open a new
dialogue box. Check the x/y-axis values; they should only differ by one decimal place. Activate the
“Scale” option (if not active by Default setting). This function will adjust e.g. the scale bar depending on
the display mode (resolution, Full screen). Enter a New Sign Name to refind your calibration easily within
the calibration table. Next, click the Save button to finish calibration.

Calibration with Scale Cross

To calibrate with a scale cross, select the respective Calibration Wizard tab.
Click Load Image button to select a scale cross from your image database.
When the image is loaded, a circle will appear on the scale cross. If desired, choose a different color.
Place the center of the circle (set as "O") exactly into the center of the scale cross. Next, take a reddish
marked point ("A") in the X-direction and drag it on the horizontal line to a certain scale division, then
perform identically in Y-direction ("B"). The magnifier window under the image window may help to place
the points accurately.
Confirm the objective lens used to capture the selected image and input the actual length of "OA" in the
"Width" bar and the actual length of "OB" in the "Height" bar. Take note about the scaling of the scale bar.
Click the Calibration button to open a new dialogue box. Continue as under Calibration with

Calibration Circle.

Calibration with Scale Line

To calibrate with scale line, select the respective Calibration Wizard. This wizard performs calibration in
X- and Y-direction separately, using the same image type: a scale cross.
Click Load Image to open the image of a scale cross. Click the Horizontal tab to activate a horizontal
line overlaying the scale cross. If desired, choose a different color. Extend the line by dragging with the
left mouse button to a defined scale division. Confirm the objective lens used to capture the selected
image and input the actual length.
Please repeat this procedure for Y-direction (Vertical tab). Click the Calibration button to continue as in

Calibration with Calibration Circle.

Calibration Table

The Calibration Table shows all calibration data saved for later usage.
If the Scale check box is selected, the measurement results will stay correct, no matter which display
format/resolution is used.

Menus and tools / Language

Select a language from the drop down list.

Menus and tools / Style

4 color styles are provided, click “Style” button and select one of them to switch the interface style.

Menus and tools / View

Displays the panel status. When you select it, the panel will display in the software interface.

Menus and tools / Image / Help

The Help menu has two commands:

Help

Provide detailed instructions on how to use the program.

About

Display the program's copyright and version information.

Control Panel

When an image is opened in Motic Images Plus 3.0 ML, the following Control Panel will display inside the Edit Tab.

Control Panel / Edit / Text

The Control Panel is composed of the following tabs: Text, Paint, Measure, and Audio.
Click Text in the Control Panel to display the text editing options.

Control Panel / Edit / Paint

The Paint tab in the Edit Control Panel allows to draw geometrical figures.

The following figures can be drawn: line, rectangle, polygon, circle, ellipse, freehand. All figures can be
edited.

Click the Icon button to bring up a dialog box which provides a variety of icons to add to the
current image.

Select a desired icon and click on a position of the image to place it.
Illumination will vary a shadow-effect representing the direction of an illumination.The slider under the
circles varies the illumination angle.
Click the Rotate 90 button to rotate the icon 90° clockwise.

Control Panel / Edit / Audio

Click the Audio tab to display the Audio control panel:

Play List brings up the Playlist window, to which external audio files can be added.

Click the right mouse button in the window to display this popup menu:

To add music to the playlist, select the Add command to display the open dialog box:

Select the desired music files, click the Open button and the selected music will be added to the playlist.
If the Auto Play option in the Audio panel of the Setting dialog box has been selected, the music in the
playlist will be played automatically each time Motic Images Plus 3.0 ML is started.
To remove the music listed in the playlist, select the Remove All command from the popup menu.

The buttons refer to Start, Pause and Stop.

Clicking dedicates the audio file to an individual image file. Move the cursor onto the image and
click left to display the following box:

The buttons , and refer to Start, Pause and Stop. Click the Record button to display the
Record window with which users are able to record sound, music, etc.

Click the button to bring up the Open dialog box from which users can open existing audio files.

Control Panel / Measure

Click the Measure tab to display the Measure control panel.

From the Objective Lens drop down list, select the objective lens that was used to capture the current
image. The color of Auto Segment can be selected.

The Measure Table button opens the following window which is composed of two tabs, Measure Table
and Options. The Measure Table tab lists all measurement results.

To export the data in the Measure Table, click "Export" to bring up the following dialog box with which
users are able to save the data as *.txt or *.cvs files.

The Measure table can be edited as well:

Toolbar / Magnifier

The Magnifier is a helpful tool especially for measurements. Start and end of a line, etc. can be precisely
defined.

Click the button to increase the magnification up to 800%.and click the button to reduce it.

Preview Window

The Preview Window is located to the bottom of the image window.
It shows all images actually opened.

How to use the Popup Menus

How to use the Popup Menus.... / ...in the Image window

Click the right mouse button in the image window to display the following popup menu:

Deselect

This command deselects the selected objects.

Lock

This command locks the selected region(s) making it impossible to move it/them around.

Freeze

This command freezes the location and size of the selected figure and fixes it to the image.

Freeze All

This command freezes the location and size of all the figures and fixes them to the image.

Delete

This command deletes the selected region.

Delete All

This command deletes all regions that have not been locked or frozen.

Deselect region

This command deselects the selected regions.

Up one layer

This command brings the selected region up one layer.

Down one layer

This command brings the selected item down one layer.

Up to top

This command brings the selected item to the top layer when there are several items on the image.

Down to bottom

This command brings the selected item to the bottom layer when there are several items on the image.

Fit to Window

This command resizes the image to fit into the current window.

Full Screen

This command brings the image window into full screen view.

Scale

This command resize the current image by scale.

Properties

This command brings up the properties dialogue to define the selected object's parameters.

File Info

This command brings up the File Info dialog box which provides information about the current image.

How to use the Popup Menus... / ...in the Album window

Click the right mouse button in the upper column of the Folder panel to bring up the following popup
menu:

New

To add a sub-folder to the selected folder.

Rename

This command enables users to rename the selected folder.

Delete

This command enables users to delete the selected folder.

Set as default capture folder

This command enables users to set the selected folder as the default capture folder. Images captured
using this program will then be saved in the selected folder automatically.

Refresh (F5)

This command enables users to refresh the contents displayed in the upper column of the Folder panel.
Pressing the F5 key will refresh the contents as well.

Calibration

Any kind of measurement, scale bar or grid needs a precise calibration for a correct quantification.
Place the calibration slide (within the delivery package of your Moticam) into the raypath of the
microscope and focus one a calibration circle (in fact it’s a dot). Depending on the objective/camera
adapter/camera sensor combination, the correct dot has to be chosen. As all dots on the calibration slide

are placed in one line, it is easy just to use the x-drive of the microscope stage to move from one dot to
another. Before taking a picture, note that the selected dot should be grabbed completely, at the same
time being be as large as possible in the image field. This will improve the precision of all measurements.
Click the Calibration button in the Measure tool bar to display the corresponding toolbar.

Calibrate... / ... with Calibration Circle

Select the Calibration Wizard command. Select a calibration method by selecting the corresponding tab.
Click the Load Image button to open an in image with a calibration circle to be selected for calibration.

Click the Open button to load the selected image.

Select the magnification of the objective used for taking this picture, enter the diameter of the calibration
dot (vacuum-metallized on the calibration slide) in the corresponding box. Click "Calibration” to continue.
Check the x/y-axis values; they should only differ one decimal place. If not, the illumination is
inappropriate.
Please note: The calibration slide is intended to work with transmitted light. In incident light, the dot
appears bright. In this case please invert the image before loading for calibration.

Activate the “Scale” option (if not active by Default setting). This function will adjust e.g. the scale bar

value depending on the display mode (resolution, Full screen). Enter a New Sign Name to refind your
calibration easily within the calibration table. Next, click the Save button to finish calibration.

Calibrate... / ... with Scale Cross

To calibrate with a scale cross, select the respective Calibration Wizard.
Click Load Image button to select a scale cross from your image database.
When the image is loaded, a circle will appear on the scale cross. If desired, choose a different color.
Place the center of the circle (set as "O") exactly into the center of the scale cross. Next, take a reddish
marked point ("A") in the X-direction and drag it on the horizontal line to a certain scale division, then
perform identically in Y-direction ("B"). The magnifier window under the image window may help to
accurately place the points.
Confirm the objective lens used to capture the selected image and input the actual length of "OA" in the
"Width" bar and the actual length of "OB" in the "Height" bar. Take note about the scaling of the scale bar.
Click the Calibration button to open a new dialogue box. Continue as under Calibration with
Calibration Circle.

Calibrate... / ... with Scale Line

To calibrate with scale line, select the respective Calibration Wizard. This wizard performs calibration in
X- and Y-direction separately, using the same image type: the scale cross.
Click Load Image to open the image of a scale cross.
Click the Horizontal tab to activate a horizontal line overlaying the scale cross. If desired, choose a
different color. Extend the line by dragging with the left mouse button to a defined scale division. Confirm
the objective lens used to capture the selected image and input the actual length.

Please repeat this procedure for Y-direction (Vertical tab).

Click the Calibration button to continue as in Calibration with Calibration Circle.

When calibration is completed, click Save to close the Calibration Wizard.

The Calibration Table shows all calibration data saved for later usage.
If the Scale check box is selected, the measurement results will stay correct, no matter which display
format/resolution is used.