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Leica DMi8A Quick Guide

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Optical Microscope Quick Start Guide The following instructions are provided as a Quick Start Guide for powering up, running measurements, and shutting down Leica’s DMi8A Inverted Optical Microscope.

Table of Contents Leica DMi8A Quick Guide ........................................................................................................... 1

Optical Microscope Quick Start Guide ....................................................................................... 2 Instrument Information ......................................................................................................................... 3 Preparing the Machine ........................................................................................................................... 3

Physical Setup ...................................................................................................................................... 3 Computer Setup .................................................................................................................................... 3 Computer Setup .................................................................................. Error! Bookmark not defined.

Operating the Microscope ...................................................................................................................... 4 Navigation ............................................................................................................................................ 4 Adjusting the Filtering Parameters ....................................................................................................... 4

Getting Results ........................................................................................................................................ 6 Taking Pictures ..................................................................................................................................... 7 Editing Pictures .................................................................................. Error! Bookmark not defined.

Finishing Up ............................................................................................................................................ 8 Turning off the System ....................................................................... Error! Bookmark not defined. Before Leaving ................................................................................................................................... 10

 Warning: Although optical microscopy has no inherent danger to usage, 

only users who have received training by qualified department staff or faculty are permitted to use this equipment. Contact the DMSE for training information.  

Notice: Italicized words indicate external buttons in the real world. Bolded words indicate buttons within the computer system.  

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Instrument Information Contrast Fields: Bright Field, Dark Field, Differential Interference Contrast, Polarizing-Contrast Magnification: 5X, 10X, 20X, 50X, 100X Focus Increments: .05 µm .1 µm, .7 µm, 1.5 µm, 5.0µ Stages: 150 mm x 200 mm

Preparing the Machine Physical Setup

1) Turn on the optical microscope then turn on the computer a. The power switch is on the CTR advanced box

2) Wait a few moments for the DMi8A to self-align and go through its boot-up sequence

Computer Setup

1) Double click on the LAS V4.7 computer icon a. The icon is the Leica Application Suite as to which the

DMi8A connects to. 2) If you only see black or white on the live feed, adjust the light

intensity to see your sample. a. Small knob on the left of the optical microscope

Figure: The Main Menu for the Optical Microscope. Areas of interest have been marked with an arrow. 

Figure: The Icon for the Leica Application Suite program. This icon 

links to the software for Leica’s 

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Operating the Microscope Navigation

1) Adjust the specimen to where you want to be using the X and Y controls a. Rotary Knobs: Top = X; Bottom = Y b. Can be done on computer w/ Mic2 c. Can be done w/ manual stage adjustment

2) Adjust the Zoom Levels on the microscope a. Available magnifications: 5x, 10x, 25x, 50x, 100x

Adjusting the Filtering Parameters Depending on the properties of your sample and its environment, different combinations of camera controls and scanning fields may be more viable than others when looking for the highest photo resolution. In order to make your sample more visible and colored to the proper extent, take a look at the following variables:

Scanning Fields The different scanning fields reflect numerous forms of illumination techniques used within advanced microscopes. Incorrect illumination of the sample can lead for sophisticated microscopes to take blurry pictures even though they are more than well-equipped. Use the following table to determine which scanning field is best for your particular sample:

Scanning Fields Advantages Disadvantages Bright Field Shows the specimen dark on a light background

ꞏAllows for viewing specimens that are unstained, transparent, and have a high reflection index

ꞏ Has a low contrast index and typically needs to be stained for proper viewing

Figure: The X‐Y‐Z Controller for the microscope. The larger two knobs 

control the X‐Y axis (respectively), and the smaller knob controls the Z‐axis.  

Figure: The available magnifications for the DMi8A. The selected magnification is highlighted orange. 

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Camera Controls Despite the importance of scanning fields for proper illumination of your sample, the camera parameters are equally consequential for getting the right tone and color balance within your picture. Any large perturbations can lead to washed out greys taking over your picture with no discernible sample features. Take a look at the following to determine which parameters are best for how you want your picture to appear:

Dark Field Shows the specimens light on a dark background.

ꞏWell suited for live and unstained samples with a low reflection index or close to its environment

ꞏ Must be very strongly illuminated which may cause damage to samples

DIC Highlights the otherwise invisible features of a sample

ꞏ Very useful for observing unstained samples that need to be stained with the other scanning fields

ꞏ Enhanced light and shadow may slightly distort the appearance of the sample

Polarizing-Contrast Uses polarizing light to enhance images of birefringent materials

ꞏ Allows for qualitative analysis of anisotropic specimen. Used primarily within optical mineralogy

ꞏ Very difficult to perform any quantitative analysis. Should only be use on very specific materials

Camera Controls Increase Effect Decrease Effect

Brightness Illumination of the sample

ꞏYour sample will appear lighter

ꞏYour sample will appear darker

Figure: The list of Scanning Fields available for the microscope 

Figure: The list of exposure adjustments available. The color 

adjustment is alongside the bottom of the tab. 

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Note: Not all camera controls will be available depending on the scanning field selected. Selecting the Easy Camera button within the tab provides less user-inputted options but a simpler experience. If you are having issues restoring the camera, press the Reset camera icon.

Getting Results

Gain Signal Intensity on the detector

ꞏBrighter images with more noise

ꞏDimmer images with less noise

Saturation Intensity of the color to white

ꞏImages will have more purer colors

ꞏImages will have more washed up colors

Gamma Factor between linear and non-linear light perception

ꞏHarder to differentiate between brighter colors

ꞏHarder to differentiate between darker colors

Color Actual color of the sample as recorded by the detector

ꞏCloser to the sample’s actual color

ꞏCloser to black and white

Figure: The Navigator tab with a few annotation options for image display resizing 

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After figuring out the best field and camera parameters for your live view, taking and annotating pictures is the next step for making your work more discernible to others. The Leica Application Suite has basic tools that can be useful for the analysis of the microstructures within your sample. With methods such as X-Y-Z stitching available, intricate and complex photos can be made. If need be, post-processing image correction along with any annotations can be applied to your image for any second thoughts on the way that the image’s camera controls were captured.

Taking and Saving Pictures 1) On the right side of the window, adjust your image size to your desired dimensions

o This can be done with the Zoom In, Zoom Out, etc. icons 2) Select the Show Navigator tab. 3) In the Navigator tab, find and left click on your data folder.

o If properly selected, you should see a red dot on the folder 4) Close out of the Navigator tab and press Acquire Image

o Acquire Image is located on the lower left corner of the screen. 5) Name your file, select the type of image, and press Save.

   

Figure: The Browse tab showing both the highlighted image, directory, and image data 

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Editing Pictures 1) Within the Process tab, select the Enhance window. 2) From here, you can change the following parameters of the photo: 

Brightness, Contrast, Gamma, Hue, Saturation, Intensity, Color, Black/White Levels.  

a. For the majority of the variables, they are now represented as 1’s or 0’s to display where the original image was before correction.  

3) This tab also allows for physical changes to the photo through: a. Orientation: Allows for flipping along the X‐Y axis or a degree 

based rotation  

Analyzing Pictures 1) At the top of the screen, select the Interactive window within the 

Analysis tab 2) Here, you will see an assortment of tools as listed:  

                    

Selection Tool: Allows for the selection of any measurements on the image Distance Line Tool: Find the distance between two points on your surface Segment Line Tool: Find the distance between a multitude of points on your surface.  Area Line Tool: Find the surface areas within a user‐defined geometry Angle Tool: Find the angle between different features on your sample  Circle Tool: Creates a circle using two points to create an arc of variable length 

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Count Tool: Figure out how many of something there is on the sample Parallel Line Tool: Create parallel lines on your sample  Cross Tool: Creates a cross on your sample Shape Tool: Creates a rectangle or a triangle on your sample.   

Note: Right clicking on the tool icon will reveal more options for data‐gathering. Such as showing either the diameter of a circle or its area=  

3) Properties allows for the parameters of the tool annotations. They function very similarly to the settings within Annotating Pictures with a couple of additions.  

a. Display Labels: Allows for the displaying of elements such as units or numbers b. Label Offset: Determines how far away you want the label from the mark 

4) Within the Automatic and Grain Expert windows, you can follow the computer‐prompted steps to automatically analyze your picture and generate a report based on your preferences for processing.  

5) When completed setting up the operation, press on Run Measurement to begin the computer‐analysis and report generation.  

Example: Simple Automatic Processing in Phase Measurement  

1) Select your Image to Measure a. In the menu, choose your photo then press Append 

2) Adjust your threshold a. Using the color wheel, adjust the hue and the saturation of the colors until your 

phases are properly selected. Clicking and dragging on the curve adjusts the hue while clicking and moving on the curve adjusts the saturation.  

3) Click on Run Measurement to get information on the phase distribution in your image.               

Finishing Up

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Before Leaving 1) Reset all objective magnifications to their lowest value. 2) Make sure that all equipment is turned off

a. Power off Sequence: Optical Microscope -> Computer 3) Clean up any and all messes created by the microscope to ensure a neat environment.

Tips and Tricks Have curves in your path that you want to analyze? Hold the LMB on the mouse and move for a continuous path to see your total distance. Want to see two image parameters that aren’t normally together? Click and hold on an empty space of the menu to pop it out of its location. Press X to return it.

FAQ

There’s no image showing up on the live feed My guided stitching failed

Apertures on Side