Leica DMI3000B User manual

3
Leica DMI3000B,
DMI4000B, DMI6000B
Instructions

5
Contents
6.10 Installation and Replacement of the
transmitted Light Lamps:
107 or 107/2 Lamp Housing........................ 45
6.11 Installing the Lamp Housing Mount
and Mirror Housing)................................... 46
6.12 Installation and Replacement of
Incident Light Lamps.................................. 48
6.13 Equipping the Incident Light
Turret Disk.................................................... 52
6.14 Inserting the Front Module Slider............ 55
6.15 Installation of the Polarizer
and Analyzer................................................ 55
6.16 Optional Accessories................................. 57
6.17 Connection to the Electronics Box.......... 58
6.18 Connection to the Computer..................... 59
6.19 Connection to the Power Supply ............. 59
7. Start-up ........................................................ 60
7.1 Functional Principle .................................. 60
7.2 Switching on the Microscope.................. 64
7.3 The LeicaDisplay ........................................ 65
7.4 The Function Buttons on the Stand......... 66
7.5 The SmartMove
Remote Control Module ............................ 69
7.6 Illumination.................................................. 69
7.6.1 Transmitted light .............................. 69
7.6.2 Incident Light - Fluorescence........ 73
7.7 Checking Phase Contrast Rings............... 74
7.8 Checking modulation contrast
slit diaphragms............................................ 77
7.9 Setting the Motorized Polarizer ............... 77
7.10 Adjusting the Light Sources ..................... 78
Contents
1. Important Notes about this Manual........ 7
2. Intended Purpose of the Microscope..... 8
3. Safety Notes................................................ 9
3.1 General Safety Notes ................................ 9
3.2 Electrical Safety.......................................... 10
3.3 Safety Instructions
for Handling the Light Sources ................ 12
3.4 Notes on handling laser devices ............. 12
3.5 Safety Instructions
for Handling Acids and Bases.................. 12
3.6 Disposal........................................................ 13
4. Overview of the Leica DMI Series.......... 14
5. Unpacking the Microscope ..................... 27
6. Assembling the Microscope.................... 30
6.1 Assembly Tools........................................... 30
6.2 Installation of the Transmitted Light
Illumination Carrier (TL)............................. 31
6.3 Installation of the DIC Module
and DIC Objective Prisms ......................... 32
6.4 Installation of Specimen Stages.............. 33
6.5 Installation of Condensers........................ 38
6.6 Installation of Eyepieces........................... 43
6.7 Installation of Objectives .......................... 43
6.8 Installation of Filters
in the Illumination Arm............................... 44
6.9 Installing the transmitted Light
Lamp Housing ............................................. 44

6
Contents
8. Operation..................................................... 81
8.1 Switching on................................................ 81
8.2 Contrast Methods....................................... 83
8.2.1 Bright Field (TL)................................ 83
8.2.2 Phase Contrast (TL) ....................... 85
8.2.3 Dark Field (TL) .................................. 86
8.2.4 Polarization (TL)............................... 87
8.2.5 Differential
Interference Contrast (TL) ............. 88
8.2.6 Integrated Phase Contrast (TL)..... 89
8.2.7 Integrated
Modulation Contrast (TL)................ 90
8.3 Fluorescence............................................... 91
8.4 Combination Methods ............................... 93
8.5 Focusing....................................................... 94
8.6 Tubes ............................................................ 96
8.7 Port selection ............................................. 96
8.8 Eyepieces..................................................... 97
8.9 Objectives.................................................... 98
8.10 Stages and Object Displacement ............ 101
8.11 Magnification Changer.............................. 102
8.12 Light sources............................................... 103
8.13 Aperture and Field Diaphragm ................ 104
9. Troubleshooting.......................................... 105
10. Care of the Microscope ............................ 109
10.1 Dust Cover ................................................... 109
10.2 Cleaning ....................................................... 109
10.3 Handling Acids and Bases........................ 110
11. Major Consumable
and Replacement Parts............................. 111
12. Dimensions.................................................. 112
13. Abbreviations and Pictograms................ 113
14. Index ............................................................ 115
15. EU Declaration of Conformity .................. 117

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4. Overview of the Instruments
Fig. 1 Left side, Leica DMI4000 B and DMI6000 B
1Eyepiece
2Eyepiece tube
3Top port
4Intermediate pupil interface
5 LeicaScreen
6Light intensity
7Field diaphragm
8TL/IL switching
9Aperture diaphragm
10 Focus wheel (motorized Leica DMI6000 B,
manual (fine and coarse) Leica DMI4000 B)
11 Variable function buttons
12 Left side port
13 Booster lens
(Leica DMI6000 B fluorescence microscopes only)
14 Lamp mount (fluorescence microscopes only)
15 Condenser head
16 Condenser base
17 Field diaphragm
18 Transmitted light lamp housing
19 DIC objective prism disk
1
2
3
4
5
678910111213
14
15
17
16
18
19

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4. Overview of the Instruments
Fig. 2 R ight side Leica DMI4000 B and DMI6000 B
1E-Focus buttons (Leica DMI6000 B only)
2Focus wheel (motorized Leica DMI6000 B,
manual (fine) Leica DMI4000 B)
3Variable function buttons
4Opener for drawer (fluorescence microscopes only)
5Drawer (fluorescence microscopes only)
6Right side port
7Analyzer slot
8Centering window (fluorescence microscopes only)
9Field diaphragm centering
(fluorescence microscopes only)
10
Incident light lamp housing (fluorescence microscopes
only)
11 Objective turret
12 Stage with attachable mechanical stage
12 3
4
5
6 78
12
9 10
11

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4. Overview of the Instruments
Fig. 3 Front view Leica DMI4000 B and Leica DMI6000 B
1 LeicaScreen
2 Front control panel
3Port switching
4 Top port
5Manual transmitted light filters
6Bertrand lens centering
1
2
3
4
6
5

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4. Overview of the Instruments
Fig. 4 General view Leica DMI4000 B and Leica DMI6000 B with SmartMove remote control module
Fig. 3b SmartMove remote control module
1Travel in x
2Travel in y
3 Focus
4Variable function buttons
(pre assigned at factory)
1
2
3
4
Fig. 3a Front control panel
1Fluorescence cube
2Shutter
3 100% light to all eyepieces
4 Port selection
5Magnification selection
61x tube lens
11
2543

60
7. Start-up
7. Start-up
7.1 Functional Principle (Leica DMI4000 B and Leica DMI6000 B)
1. Intelligent Automation
• Switching between contrast methods at the touch of a button. Light rings, DIC prisms, etc.
are automatically positioned in the beam path.
• The microscope recognizes the selected objective and associated contrast method.
The intensity (INT), aperture diaphragm (AP) and field diaphragm (FD) are always set to suit-
able values.
• The INT, AP and FD values are always based on the currently activated illumination axis
(transmitted light or incident light).
• The INT, AP, and FD values can be adjusted individually. Manual adjustments overwrite the
previous settings. The current setting is stored and is retained from one session to the next
when power is switched off.
2. Controls
• SmartMove knobs
for stage and focus control
• Fixed function buttons on stand
for INT, AP, and FD, as well as for switching between transmitted light and incident light axis
• Variable function buttons on stand, SmartMove, STP6000
These function buttons have functions suitable to the configuration of your microscope as-
signed to them at the factory. The functions can be reprogrammed and/or adapted to your
specific requirements, however.
• Complete control of microscope and camera via software
(Leica Application Suite (LAS/LAS AF))
Thanks to its intelligent automation, the Leica DMI4000 B and DMI6000 B can be controlled using a
variety of control elements.

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7. Start-up
Function Fixed Variable SmartMove Software
(DMI4000 B and DMI6000 B) Function Function Function Rotary and
buttons buttons buttons knobs STP6000
Stand Stand
4000 6000 4000 6000 4000 6000 4000 6000 4000/6000
Select contrast method - - + + + + - - +
Change transmitted light/incident light axis + + - - - - - - +
Change to objective - - - + - + - - +
Teach-in parfocality - - - - - - - - +
Change operating mode (dry/imm) - - + + + + - - +
Illumination Manager + + + + + + - - +
Magnification changer (motorized) + + - - - - - - +
Focusing - + - - - - - +1) +
Set stops - + - - - - - - +
Go to stop - + - - - - - - +
Change step increment (coarse/fine) - - - + - + - - +
XY stage positioning - - - + +
Change speed - - - - - - - - +
Stage positions (store/go to) - - - - - - - - +
Change to filter/reflector cube + + (+) + + - - +
Side and bottom port (DMI6000 B only) + (+) + - +
DIC fine adjustment + + - - - - - - +
Adapted Focus Control (AFC) - + - + - + - - +
+ always possible
(+) optional
- not possible
1) Focusing alternatively via wheels

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7. Start-up
Possible Assignments for Variable Function Buttons on Stand and SmartMove
For Leica DMI4000 B and Leica DMI6000 B:
Function button Function
BF Bright field transmitted light
PH Phase contrast transmitted light
ICT Interference contrast, transmitted light
DF Dark field transmitted light
IMC Integrated modulation contrast
POL Polarization transmitted light
CHANGE TL Cycle through all contrast methods
INT ↑ Increase intensity (transmitted light)
INT ↓ Reduce intensity (transmitted light)
AP ↑Open aperture diaphragm (transmitted light)
AP ↓ Close aperture diaphragm (transmitted light)
FD ↑ Open field diaphragm (transmitted light)
FD ↓ Close field diaphragm (transmitted light)
SHUTTER TL Open/close TL shutter
TL FLT 1 Enable/disable transmitted light filter at position 1
TL FLT 2 Enable/disable transmitted light filter at position 2
FLUO Fluorescence (last filter cube)
CUBE 1-6 Select filter cube in position 1-6
CHANGE CUBE CW Change cube clockwise (1 → 4)
CHANGE CUBE CCW Change cube counterclockwise (4 → 1)
INT FLUO ↑ Increase intensity (fluorescence)
INT FLUO ↓ Reduce intensity (fluorescence)
FD FLUO ↑ Open field diaphragm (fluorescence)
FD FLUO ↓ Close field diaphragm (fluorescence)
CHG FW Toggle filter functions
IFW Activate external filter wheel
ExMan Activate Excitation Manager
SHUTTER FL Open/close fluoshutter
COMBI Combination method (PH fluorescence or ICT fluorescence)
CHANGE COMBI Cycle through all combination methods
CHANGE OBJ CW Cycle through objectives clockwise
CHANGE OBJ CCW Cycle through objectives counterclockwise
Z FINE Activate fine focusing (Leica DMI6000 B only)
Z COARSE Activate coarse focusing (Leica DMI6000 B only)
XY PRECISE Activate precise stage
XY FAST Activate fast stage
BTP ON/OFF Bottom port on/off (Leica DMI6000 B only)
DRY/IMM Switch dry/immersion
CHANGE FLT Switch TL filter
CHANGE CS Switch to confocal application
OBJ 1-6 Select objective at position 1-6
MEM 1-6 Memory activated stored functions
AFC ON/OFF Turns AFC on or off
AFC HOLD Holds current position

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7. Start-up
If a component has not been installed correctly,
the LeicaScreen will display an error message.
See Troubleshooting chapter, → p. 105.
Components such as diaphragms, condensers,
light, and phase rings have been pre-centered
at the factory. It may be necessary to correct the
centering after the microscope has been trans-
ported and assembled.
Before performing the required steps, please fa-
miliarize yourself with the LeicaScreen and the
controls.
Caution!
After turning on the gas discharge lamp, the
burner must be immediately adjusted. There-
fore, do not turn on the power supply unit
yet. First, work in transmitted light in order
to familiarize yourself with the microscope’s
controls.
Fig. 81
LeicaScreen
Initialization
7.2 Switching on the Microscope
Leica DMI3000 B:
• Switch on the microscope’s power at the On/
Off switch. The signal lamp is lit when the in-
strument is ready. (For the Leica DMI3000 B
please continue at 7.4. Function Buttons on the
Stand)
Leica DMI4000 B and Leica DMI6000 B:
• Switch on the power of the electronics box at
the On/Off switch (80.1). The signal lamp (80.2)
is lit green when the unit is ready. All motorized
microscope components will then run through
an initialization phase.
Note:
If a PC is connected, switch on the electronics
box first, and then the computer.
After the initialization (Fig. 81) is complete, the
LeicaScreen will display the microscope’s cur-
rent settings (Fig. 82).
Fig. 80
front side
Leica CTR6000
1On/Off switch
2Signal lamp
1
2
Fig. 82
LeicaScreen
after
Initialization

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7. Start-up
7.3 The LeicaDisplay
(Leica DMI 4000 B and DMI 6000 B)
The screen displays the microscope’s current
settings. The content of the display depends on
the features of the individual microscope.
For information on the abbreviations used, please
turn to the table of abbreviations → p. 113.
The screen has a number of areas and lines.
Line 1: contrast method
Line 2: objective/magnification
Line 3: illumination/diaphragms
Line 4: active ports
Line 5: focus/stops
(DMI 6000 B only)
The content of the display changes according to
the active function.
Contrast method
Objective/
Magnification
Focus/stops
(DMI6000 B only)
Ports/Eyepiece
Illumination
Diaphragm
Fig. 83 Arrangement of the function buttons – overview
1Four variable function buttons
2Illumination Manager
3 Front control panel
4Focus buttons (DMI6000 B only)
5Three variable function buttons
6SmartMove knobs
7SmartMove function buttons
543321
6
77
6
Pictograms

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7. Start-up
Note:
Changes to the light intensity as well as aperture
and field diaphragm settings are stored for the in-
dividual objectives and contrast methods.
Fig. 84 Fixed function buttons (left side of stand)
1Toggle transmitted light/incident light
2 Aperture diaphragm
3light intensity
4field diaphragm
4
1
3
2
7.4 The Function Buttons on the Stand
Leica DMI3000 B:
• Focus wheels: the left-hand focus wheels can
be used for both coarse and fine focusing; the
right-hand focus wheel for fine focusing only
(a version of the Leica DMI3000 B with mir-
rored focus controls is also available)
• Light intensity: the transmitted light intensity
can be adjusted continuously from 0 to 12 V
using the potentiometer at the lower left of the
front of the microscope stand.
For the Leica DMI3000 B please continue at 7.6.
Illumination.
Leica DMI4000 B and Leica DMI6000 B:
A number of function buttons are located on both
sides of the stand. These can be broken down
into fixed and variable buttons. The variable func-
tion buttons have different functions depending
on the features of the individual microscope.
Fixed function Buttons on the Left Side
The TL/IL button (84.1) toggles between the inci-
dent-light and transmitted light axis. The contrast
method last used with a given axis is restored
when switching.
The INT buttons (84.3) adjust the light intensity.
The adjustment can be made in coarse or fine
steps. Pressing both INT buttons at the same time
toggles between coarse and fine adjustment.
The AP buttons (84.2) for the aperture diaphragm
and FD (84.4) for the field diaphragm open and
close their respective diaphragms.
The motorized DMI stands can be controlled
using the function buttons on the stand, the re-
mote control SmartMove or the STP6000.

67
7. Start-up
Variable Function Buttons on the Stand
The variable function buttons are assigned func-
tions at the factory that are appropriate to the
features of your microscope. They are labeled
accordingly. For details on button assignments,
please refer to the included identification sheet.
For information on the abbreviations used, please
refer to the list → p. 63.
Note:
The Leica Application Suite (LAS) software is re-
quired for changing the button assignments.
Fig. 85 Function buttons (left side of stand)
1Variable function buttons
2Open/close aperture diaphragm
3TL/IL switching
4Open/close field diaphragm
5Increase/decrease light intensity
Fig. 86 Function buttons (right side of stand)
1Variable function buttons
BF
PH
ICT
DF
IMC
POL
CHANGE TL
INT ↑
INT ↓
AP ↑
AP ↓
FD ↑
FD ↓
SHUTTER TL
TL FLT 1
TL FLT 2
FLUO
CUBE 1
CUBE 2
CUBE 3
CUBE 4
CUBE 5
CUBE 6
AFC ON/OFF
AFC HOLD
CHANGE CUBE CW
CHANGE CUBE CCW
INT FLUO ↑
INT FLUO ↓
FD FLUO ↑
FD FLUO ↓
CHG FW
IFW
ExMan
COMBI
CHANGE COMBI
CHANGE OBJ CW
(only DMI6000 B)
CHANGE OBJ CCW (only DMI6000 B)
Z FINE (only DMI6000 B)
Z COARSE (only DMI6000 B)
XY PRECISE
XY FAST
BTP ON/OFF
(only DMI6000 B)
DRY/IMM
CHANGE FLT
CHANGE CS
OBJ 1-6
MEM 1-6
Turns AFC on or off
Holds current position
Possible functions*:
* See page 63 for abbreviations
54321
1

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7. Start-up
Function Buttons on the Front Panel (Fig. 87)
100% of the light goes to the eyepiece
(87.1).
Toggle function for the side ports
(87.2). This function depends on the
individual microscope configuration.
Note:
Switching to the bottom port:
via the variable function buttons (Lei-
ca DMI6000 B only), switching to top
port: manually.
SHUTTER Opens and closes the shutter (87.3).
Switches between the possible
magnifications of the magnification
changer (87.4).
The magnification changer is set to
the magnification 1x (87.5).
Fig. 87 Front control panel
1100% light to eyepiece
2Toggle ports
3Shutter
4Switch between subsequent magnifications
5Subsequent magnification 1x
6Selecting filter cubes
Fig. 88
1Focus control buttons
2Open filter drawer
CUBE The CUBE 1 to CUBE 6 (87.6) buttons
permit the direct selection of indi-
vidual filter cubes, provided the se-
lected cube is valid for the selected
method.
Press the CUBE 3 and CUBE 4 but-
tons at the same time to display the
assignments of the variable function
buttons. To reset the display, press
the buttons again or wait 3 seconds.
Focus buttons (Fig. 88) (DMI6000 B only)
Moves the Z drive in the indicated di-
rection.
SET + Z↑Sets the upper focus stop.
SET + Z↓Sets the lower stop.
1
Z↑
Z↓
2
1x
→
→
→
→
13
4
66
2 5

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7. Start-up
Fig. 89 SmartMove remote control module
1travel in x
2Travel in y
3 Focus
4Individual adjustment of button height
5Variable function buttons (factory preset)
1
2
3
4
5
Shutter button + Cube buttons 1-6 (Leica
DMI4000 B and Leica DMI6000 B only)
The selected cube is moved to the
loading position for replacement.
„Load“ appears on the screen. After
pressing the button (88.2) the draw-
er is opened and the cube can be
changed. The next filter cube will be
moved to the loading position after
the drawer is closed.
7.5 The SmartMove Remote Control Module
SmartMove knobs
(Leica DMI4000 B and Leica DMI6000 B)
Use the knobs 89.1 and 89.2 to move the stage in
X and Y directions.
The image is focused using the knob 89.3 (Leica
DMI6000 B only).
The height of the knobs can be adjusted to a
comfortable working position by turning 89.4.
Variable function buttons on SmartMove
The variable function buttons are assigned func-
tions at the factory that are appropriate to the
features of your microscope. They are labeled
accordingly. For details on button assignments,
please refer to the included identification sheet.
For information on the abbreviations used, please
refer to the list → p. 63.
Note:
The Leica Application Suite (LAS) software is re-
quired for changing the button assignments.
7.6 Illumination
7.6.1 Transmitted light
If your microscope has not yet been set up for
Koehler illumination, please continue with the
„Koehler Illumination“ section.
Leica DMI3000 B:
• If necessary, adjust the TL bright field position
at the filter slider.
• Select an objective with moderate magnifica-
tion
(10x–20x) .
• Set the condenser to the bright field position.
• Place a specimen on the stage.
• Focus on the specimen using the focus wheels.
• Adjust the light intensity.
• Close the field diaphragm manually until the edge
of the diaphragm appears in the field of view.

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8. Operation
8.2 Contrast Methods
All of the contrast methods of the Leica DMI4000
B and Leica DMI6000 B can be selected and con-
trolled via the variable function buttons and the
Leica Application Suite (LAS). The only excep-
tions are methods that involve components re-
quiring manual control (e.g. systems with manual
analyzers). The following section describes the
use of the function buttons on the stand. For in-
structions on the use of the software, please re-
fer to the separate manual.
Contrast methods for the Leica DMI3000 B are
controlled via the manual condenser, the manual
objective turret, as well as turning knobs and
sliders at the microscope.
8.2.1 Bright Field (TL)
Leica DMI3000 B:
• If necessary, adjust the TL bright field position
at the filter slider.
• Set the condenser to the bright field position.
• Remove all other optical components such
as analyzers, polarizers or IC prisms from the
beam path.
• Insert a transmitted light specimen.
• Select your objective
• Set the brightness at the light potentiometer
• Focus the image with the focus wheels.
Leica DMI4000 B and Leica DMI6000B:
Note:
If all positions of the filter turret are occupied, fil-
ter cube „A“ can be swapped for filter cube „A-
TL“ using the Leica Application Suite (LAS/LAS
AF). TL contrast methods are possible with that
filter cube.

84
8. Operation
• Use the TL/IL function button to switch to
transmitted light (TL).
• Select the BF (bright field) contrast method
by pressing the variable button BF.
Alternatively: press the variable button
CHANGE TL .
(For details on button assignments, please see
the identification sheet.)
BF will appear on the LeicaScreen.
Motorized condensers will now move to the
bright field position. Coded condensers must
be switched manually.
The fluorescence filter turret will automatically
go to an empty position or to the „A-TL“ filter
cube.
Fig. 108 Function buttons (left side of stand)
1variable function buttons
2Open/close aperture diaphragm
3TL/IL switching
4Open/close field diaphragm
5Increase/decrease light intensity
Fig. 109 Function buttons (right side of stand)
1variable function buttons
54321 1
• Insert a transmitted light specimen.
• Rotate an appropriate objective into place.
• Focus the image with the knob on the Smart-
Move or the focusing wheel and adjust the in-
tensity with the INT function buttons.

85
8. Operation
8.2.2 Phase Contrast (TL)
(integrated phase contrast, see 8.2.6)
Leica DMI3000 B:
• If necessary, adjust the TL bright field position
at the filter slider.
• Select a phase contrast objective.
• Select the suitable light ring on the condenser.
• Open the aperture of the condenser com-
pletely.
• Remove all other optical components such
as analyzers, polarizers or IC prisms from the
beam path.
• Insert a phase contrast specimen.
• Set the brightness at the light potentiometer
•Focus the image with the focus wheels.
Leica DMI4000 B and Leica DMI6000 B:
• Use the TL/IL function button to switch to
transmitted light (TL).
• Select the PH (phase contrast) contrast meth-
od
by pressing the variable button PH.
Alternatively: press the variable button
CHANGE TL .
(For details on button assignments, please see
the identification sheet.)
PH will appear on the LeicaScreen.
Motorized condensers will now switch to the
correct light ring. Coded condensers must be
switched manually.
• Insert a transmitted light specimen.
• Rotate an appropriate objective into place.
Objectives that are suitable for phase contrast
are engraved with PH.
• Focus the image with the knob on the Smart-
Move or the focusing wheel and adjust the in-
tensity with the INT function buttons.
Note:
When selecting the phase contrast method, the
aperture diaphragm is opened fully and can not
be adjusted.

91
8. Operation
8.3 Fluorescence
Leica DMI3000 B:
The filter slider (5a.9, S.24) is used to operate the
fluorescence module.
• Pull the filter slider out completely to open the
beam path.
• Push the filter slider into the middle position
(1st detent) to bring the blue filter into the
beam path.
• Insert the filter slider fully in order to block the
beam path (shutter position).
• The fluorescence illumination is controlled by
the rotary knob (5a.10, S.24).
• The filter cubes are swiveled manually into the
beam path by turning the incident light turret
disk.
Leica DMI4000 B and Leica DMI6000 B:
• Use the TL/IL function button to switch to fluo-
rescence FLUO.
• Place a specimen on the stage and select a
suitable objective.
• The current fluorescence filter cube will be
displayed on the LeicaScreen.
• You may protect your specimen from fading by
closing the incident light shutter.
To do so, press the SHUTTER button (87.3) on
the front panel.
The following pictogram will appear on th Lei-
caScreen:
Changing the fluorescence filter cube
Fixed function buttons on the front panel:
CUBE 1 to CUBE 6 or Cube CCW
Variable function buttons on the front panel
and SmartMove: CUBE CW or CUBE CCW
Leica Application Suite (LAS) Software
• Focus the image with the knob on the Smart-
Move or the focusing wheel and adjust the in-
tensity with the INT function buttons.