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Leica DMI 4000 tutorialBefore using the Leica

DMI 4000,

You will need to put down your name on the reservation system =

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Welcome to the Leica DMI4000 Microscope tutorial

How to start up the system (p.3)How to use the stand DMI4000 (p.5)How to use the stand from the soft (p.10)The Differential Interference Contrast (DIC) (p.11)Fluorescence (p.12)How to acquire an image and its different adjustments (p.13)How to focus easier (p.17)How to acquire several images and overlay them (p.18)How to see, to save your images and their information (p.21)How to annotate your image (p. 22)How to shut down the system (p.23)

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System Start up (step1)

Turn on the Fluorescence lamp (Xcite)

Turn on the Microscope button

You have to turn on the light only if it is cold (approximately 30 minutes after its last use)

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System Start up (step2)Turn on the PC

Introduce your UsernameIntroduce your password

Start the Leica Application suite software

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Microscope ( left side)

Intensity of halogen or fluorescence lamp (+/-) (INT)

Condenser Aperture for the contrast (+/-) (AP)

Fluorescence to transmitted lightselection

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Microscope (front part)Eyes piece position

Camera position

Shutter for the fluorescence (Xcite)

Filters (BLUE/GREEN/RED/DEEP RED)

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Microscope (right side)Choose your objective manually:

HCX PL S-APO 5x 0.15

HCX PL APO 10x 0.30

N PLAN 10x 0.25

HC PL APO 20x 0.7 IMM CORR λbl

HCX PL APO 63x 1.40-0.6 OIL

To put a magnifying lens (1.6 x)

To move the stage

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Microscope (up part)

Field diaphragm, for Köhler adjustment (close/open)

condenser

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Check your sample through the eyes piece

Select the objective

For bright field

On the left side, Press on the TL/BF buttonSetup the intensity, with the buttons INTsetup the aperture diaphragm, with the button APFocus the sample

For fluorescence

On the front part,Select the filter (BLUE / GREEN / RED / DEEP RED )Press the shutterFocus the sample

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Microscope on the softUnder Acquire and Mic1,

you can see the objective that you’re using

You can change the kind of contrast in transmitted light, or select fluorescence

For Fluorescence, you can change filters and press the shutter

You can select the eye piece or camera position

For Acquiring your image after all settings done

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Differential interference contrast (DIC)

Under Acquire and MIC1, select a DICcontrast

You can change the aperture diaphragm and the intensity of the light

To change and choose the right contrast , turn this button

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FluorescenceUnder Acquire and MIC1, select

FLUO, so you can

setup the field diaphragm

Setup the intensity of the fluorescent light

Change the filters

Press the shutter

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Acquire an imageUnder Acquire and Camera, you can adjust

the exposure time

The contrast (gamma)

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Histogram and saturation optionsUnder Acquire and Camera and Histogram,

You can see the grey scale

You can tick Show Over/Under Exposure, and you will see the over exposure in red and the under exposure in blue, you shouldn’t have some red dots on your picture

You can also reduce the grey scale

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The Input optionsUnder Acquire and camera and input

option, you can choose

if you want some pseudo color

The bit depth

The right format (HQ 1600x1200)

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The binning

Under Acquire and Camera and Input option,

You can choose a binning, it will increase the camera sensitivity, reduce the exposure time

But you will lose some resolution

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How to focus easierUnder Acquire and Camera

and Region of interest,

If you select zoom focus, it will zoom the region you want so you can easily find the right focus

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How to acquire several images and overlay them

Under Select Acquisition Mode, select Image Overlay

The window “lambda” will be opened

With the play button, you can start and stop the live and the shutter

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To do the overlay and save the configuration

For each channelyou have to select the filterChoose the exposure timeChoose a pseudo-color

You can save your configuration

When it’s done you can Acquire your overlay

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To see the overlay imageUnder Browse and λ, you can see

Each channelAnd the overlay picture

You can also tick the images that you want to see in overlay mode and click on overlay show

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How to see, to save your images and their information

Under Browse and file information, you can see the information linked to the image

Under Browse and File directory, you can see in which file your images are saved

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How to annotate your imageUnder Process and Annotate, you can choose to note some information about the image

The name , description , date and time

The scale bar

Lines, measurements (ex. distance line)

To save the picture with the information, you have to select Merge.

Be careful, if you do it, you can’t go back. So if you need the original picture, copy it before you annotate it

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How to shut down the system

If you was using oil/glycerol objective, clean the objective with ethanol/water

Before shutting down the system, you have to look if there is somebody using this microscope after you. In this case you haven’t to shut it down but you have to log off your session

Shut down the PCTurn off the microscopeTurn off the light