Leica DMIRE2 User manual

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Cell Imaging Unit – UIC
LEICA DMIRE2 MICROSCOPE MANUAL

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TABLE OF CONTENTS
ANATOMY OF THE LEICA DMIRE2 INVERTED MICROSCOPE 3
INITIALISATION PROCEDURE 4
LEFT SIDE OF MICROSCOPE (DETAILED VIEW) 5
DMIRE2 FRONT PANEL 6
1. LCD DISPLAY OF MICROSCOPE STATUS 6
2. LCD DISPLAY CONTROL BUTTONS 7
3. L.E.D. PANEL SHOWING ACTIVE FLUORESCENCE CUBE 7
4. FILTER CUBE SELECTION SWITCH 7
5. MERCURY LAMP SHUTTER 8
6. PORT SELECTION SWITCH 8
7. TUBE LENS 8
CONTRAST METHODS 9
BRIGHTFIELD 9
FLUORESCENCE 9
MAGNIFICATION 11
CHANGING OBJECTIVE 11
FOCUSSING 12
SCANNING MODE 13

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ANATOMY OF THE LEICA DMIRE2 INVERTED MICROSCOPE
1. Binocular phototube 16. Filters
2. Eyepiece adapter tube 17. Aperture diaphragm adjustment
3. Eyepieces 18. Lamp housing mount
4. Tube mount 19. Lamp housing
5. Tube port for photo/TV connect 20. Stage plate
6. Beam splitter 21. Analyser
7. Mains switch 22. Tube lens module
8. Brightness adjustment 23. Switch rod for lateral TV port
9. Lateral TV port 24. Transmitted light illumination
10. Coaxial coarse & fine focus 25. Condenser
11. Fluorescence module 26. Transmitted light lamp housing
12. ICT prism adjustment 27. Transmitted light field diaphragm
13. Sextuple objective nosepiece 28. SLR port
14. Centring buttons for incident light 29. Second lamp housing
15. Field diaphragm adjustment

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INITIALISATION PROCEDURE
1. Ensure that both the DMIRE2 control box (CTRMIC) and the mercury arc lamp
power supply have been switched on (shown below). After switching on the
CTR6000 control box the microscope will take a few seconds to initialize
2. During microscope initialisation the stage position will be reset and recently used
settings will be recalled. After initialisation the microscope status menu will appear
on the L.C.D. display screen (shown below).

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LEFT SIDE OF MICROSCOPE (DETAILED VIEW)
A Halogen lamp intensity adjustment wheel
B Coaxial coarse & fine focus
*Beam splitter (not functional)
C Epifluorescence illumination diaphragm (controls brightness of illumination)
DEpifluorescence field diaphragm (controls area of illumination)
EObjective prism turret
FFilter cube turret

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DMIRE2 FRONT PANEL
1. LCD DISPLAY OF MICROSCOPE STATUS (ALPHA-NUMERIC).
Displays the following information:
•Position of objective lens relative to upper limit (e.g. -145µm)
•Step size setting for focussing (S0-3)
•Upper & lower limits for stage position (↑↓)
•Current objective settings (5x, 10x, 25x, 40x, 63x)
•Current filter cube. For brightfield (BF) or fluorescence optics (A, N21, I3; below)
•Dry/ immersion mode (D/I)

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2. LCD DISPLAY CONTROL BUTTONS
LEARN Puts microscope in learn mode (DO NOT USE). If you press
this button by accident, you will see the word “EXIT” flashing.
Press the LEARN button again to exit the learn mode.
CHANGE Toggles between voltage readout and objective readout.
Set lower limit of travel for objective (DO NOT USE)
Set upper limit of travel for objective (DO NOT USE)
STEP Adjust step size for coarseness of focussing. Four settings are
available (S0, fine; S1, medium fine; S2, medium coarse & S3,
coarse).
3. L.E.D. PANEL SHOWING ACTIVE FLUORESCENCE FILTER CUBE
FILTER COLOUR OF LIGHT FLUOROCHROMES
(1) --------------- ---------------
(2) DAPI 31000UZ
UV light (BP350/50) DAPI
(3) GFP
Blue light (BP470/40) FITC, Alexa 488
(4) mRFP
Green light (BP 584/50) TRITC, Texas Red, Alexa 546
4. FILTER CUBE SELECTION SWITCH (TWO-WAY)
Pressing the left button will rotate the motorised filter cube turret left, whereas the
right switch will rotate the filter cube turret right. The active filter cube will be
illuminated (red) in the horizontal LED panel (see above).
5. MERCURY LAMP SHUTTER SWITCH (ONE WAY)
Red LED is on when shutter is closed

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6. PORT SELECTION SWITCH (TWO WAY)
VIS Light directed to binocular eyepieces
SIDE Light directed to scanner
BOTTOM Disabled
The active port will be illuminated (red) in the vertical LED panel.

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CONTRAST METHODS
BRIGHTFIELD
Use the wheel on the front left side of the microscope stand near the base to turn on
and adjust the halogen lamp brightness (shown below).
The lamp voltage will be displayed automatically on the microscope control panel
when the intensity is adjusted. To switch off transmitted light illumination adjust lamp
intensity to 2.5V then continue rotating the wheel beyond this point. 0V on control
panel display indicates illumination is off. To switch on, rotate the wheel in the
opposite direction.
FLUORESCENCE
Ensure transmitted light has been switched off (0V on LCD display panel) and the
mercury lamp is switched on. Cycle through fluorescent cubes by pressing the
fluorescent cube selection switches (*).
Press the shutter switch to close the mercury lamp shutter (red light is on when shutter
is closed).

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MAGNIFICATION
The following objective lenses are fitted on the DMIRE2:
TUR
Ret
pos
Objective type Mag N.A. Immersion Cover-glass
DIC prisms
(upper S1)
DIC
prisms
(upper
S23)
DIC
prisms
(lower)
Pixel size
(1xc-
mount)
(um/pixl)
1 N PLAN 2,5x 0.07 DRY With/without -K1 C 2.58
2 HC PL FLUOTAR 10x 0.30 DRY With /without Not avail. (K11) K3 A 0.64
3 HC PL APO 20x 0.70 DRY 0.17 #1.5 K3 K3 D
4 HCX PL APO 40x 0.85 DRY 0,17 #1.5 K3 K7 A 0.16
5 HCX PL APO CS 63x 1.30 GLYC 0.17 #1.5 K7 K7 D
6 HCX PL APO 100x 1.40-
0.70 OIL 0,17 # 1.5 K7 Not avail. (K10)
D
HCX APO
* 40x 0.80 WATER With /without - - - -
CHANGING OBJECTIVE
To reduce the chance of immersion medium getting on to a dry objective, the
microscope will not allow you to move freely between oil immersion and dry lens
mode. To switch from one mode to another, simultaneously press the upper limit and
lower limit buttons on the microscope control panel (asterisks).
The words “CHANGE OBJECTIVE” will flash on the LCD display. Now you can
change the objective using the objective turret control buttons located behind the
coaxial coarse and fine focus controls on the LEFT side of the microscope (upper
button increases the magnification; lower button decreases the magnification)

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FOCUSSING
1. Coaxial focus knobs
Coaxial focussing knobs are located on the left and right side of the microscope.
Turning the knob so that your thumb moves away from you focuses down. Turning
the knob so that your thumb moves toward you focuses up.
2. Focussing buttons
In addition to using the conventional coaxial focussing mechanism, it is possible to
use the focussing buttons located on the right side of the microscope (behind coaxial
focus control) to focus your specimen. The upper key will focus up whereas the lower
key will focus down.
If an upper limit is set for the objective (See DMIRE2 Front Panel – LCD Display
Control Buttons) then the objective will not move above that limit when using the
focussing buttons. The only way to focus above the upper limit is to use the focusing
knobs.*
The focussing mechanism is electronic and has five settings:
Setting Step Size
S0 Fine
S1 Medium fine
S2 Medium coarse
S3 Coarse
* This is a safety feature to prevent accidental damage of the objective.

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SCANNING MODE
1. If you didn’t already does it start the Image-Pro Plus software available both
as a Desktop shortcut and the Windows start menu.
2. On Image-Pro Plus press the small camera in the toolbar for acquisition.
3. On the Acquire dialog:
o Press “Preview” to preview your image.
o Adjust the exposure time with the small bars in the center of the
window for optimal contrast.
o When you’re happy with your image press “Snap” to get the image.
4. Save the image to your folder on D:\users.