Leica DMi8A Manual de usuario

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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

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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.

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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

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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:

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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

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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.

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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=

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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

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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.

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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

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