Leica Microsystems

DMI3000 B, 4000 and B 6000 Instructions

Instructions

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File Name: Leica Microsystems - DMI3000 B, 4000 and B 6000 Instructions.pdf

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Leica DMI3000 B, DMI4000 B, DMI6000 B Instructions

Contents

Contents 1.

Important Notes about this Manual ...

Intended Purpose of the Microscope ...

3. 3.1 3.2 3.3

9 9 10

3.6

Safety Notes ... General Safety Notes ... Electrical Safety... Safety Instructions for Handling the Light Sources ... Notes on handling laser devices ... Safety Instructions for Handling Acids and Bases ... Disposal...

Overview of the Leica DMI Series ... 14

Unpacking the Microscope ... 27

6. 6.1 6.2

Assembling the Microscope ... Assembly Tools ... Installation of the Transmitted Light Illumination Carrier (TL) ... Installation of the DIC Module and DIC Objective Prisms ... Installation of Specimen Stages ... Installation of Condensers ... Installation of Eyepieces ... Installation of Objectives ... Installation of Filters in the Illumination Arm... Installing the transmitted Light Lamp Housing ...

3.4 3.5

6.3 6.4 6.5 6.6 6.7 6.8 6.9

12 12 12 13

30 30 31 32 33 38 43 43 44 44

6.10 Installation and Replacement of the transmitted Light Lamps: 107 or 107/2 Lamp Housing... 6.11 Installing the Lamp Housing Mount and Mirror Housing) ... 6.12 Installation and Replacement of Incident Light Lamps ... 6.13 Equipping the Incident Light Turret Disk... 6.14 Inserting the Front Module Slider ... 6.15 Installation of the Polarizer and Analyzer... 6.16 Optional Accessories... 6.17 Connection to the Electronics Box ... 6.18 Connection to the Computer ... 6.19 Connection to the Power Supply ... 7. 7.1 7.2 7.3 7.4 7.5

Start-up ... Functional Principle ... Switching on the Microscope ... The LeicaDisplay ... The Function Buttons on the Stand ... The SmartMove Remote Control Module ... 7.6 Illumination ... 7.6.1 Transmitted light ... 7.6.2 Incident Light - Fluorescence... 7.7 Checking Phase Contrast Rings ... 7.8 Checking modulation contrast slit diaphragms... 7.9 Setting the Motorized Polarizer ... 7.10 Adjusting the Light Sources ...

45 46 48 52 55 55 57 58 59 59 60 60 64 65 66 69 69 69 73 74 77 77 78

Contents 8. 8.1 8.2

Operation ... 81 Switching on... 81 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

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

4. Overview of the Instruments

1 16 2 15 3

Fig. 1 Left side, Leica DMI4000 B and DMI6000 B 1 Eyepiece 2 Eyepiece tube 3 Top port 4 Intermediate pupil interface 5 LeicaScreen 6 Light intensity 7 Field diaphragm 8 TL/IL switching 9 Aperture diaphragm 10 Focus wheel (motorized Leica DMI6000 B, manual (fine and coarse) Leica DMI4000 B)

9 8 7

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

4. Overview of the Instruments

4 5

Fig. 2 Right side Leica DMI4000 B and DMI6000 B 1 E-Focus buttons (Leica DMI6000 B only) 2 Focus wheel (motorized Leica DMI6000 B, manual (fine) Leica DMI4000 B) 3 Variable function buttons 4 Opener for drawer (fluorescence microscopes only) 5 Drawer (fluorescence microscopes only) 6 Right side port 7 Analyzer slot

7 8

8 9

Centering window (fluorescence microscopes only) Field diaphragm centering (fluorescence microscopes only) 10 Incident light lamp housing (fluorescence microscopes only) 11 Objective turret 12 Stage with attachable mechanical stage

4. Overview of the Instruments

6 3

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

4. Overview of the Instruments Fig. 3a Front control panel 1 Fluorescence cube 2 Shutter 3 100% light to all eyepieces 4 Port selection 5 Magnification selection 6 1x tube lens

3 4

Fig. 3b SmartMove remote control module 1 Travel in x 2 Travel in y 3 Focus 4 Variable function buttons (pre assigned at factory)

5 3

1 2

Fig. 4

General view Leica DMI4000 B and Leica DMI6000 B with SmartMove remote control module

7. Start-up

7. Start-up 7.1 Functional Principle (Leica DMI4000 B and Leica DMI6000 B) Thanks to its intelligent automation, the Leica DMI4000 B and DMI6000 B can be controlled using a variety of control elements. 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 suitable 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 assigned 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))

7. Start-up

Function (DMI4000 B and DMI6000 B)

Fixed Function buttons Stand

Variable Function buttons Stand

SmartMove Function Rotary buttons knobs

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 Set stops Go to stop Change step increment (coarse/fine)

+ + + -

+1) -

+ + + +

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

Focusing alternatively via wheels

Software and STP6000

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 PH ICT DF IMC POL CHANGE TL

Bright field transmitted light Phase contrast transmitted light Interference contrast, transmitted light Dark field transmitted light Integrated modulation contrast Polarization transmitted light Cycle through all contrast methods

INT ↑ INT ↓ AP ↑ AP ↓ FD ↑ FD ↓ SHUTTER TL TL FLT 1 TL FLT 2

Increase intensity (transmitted light) Reduce intensity (transmitted light) Open aperture diaphragm (transmitted light) Close aperture diaphragm (transmitted light) Open field diaphragm (transmitted light) Close field diaphragm (transmitted light) Open/close TL shutter Enable/disable transmitted light filter at position 1 Enable/disable transmitted light filter at position 2

FLUO CUBE 1-6 CHANGE CUBE CW CHANGE CUBE CCW

Fluorescence (last filter cube) Select filter cube in position 1-6 Change cube clockwise (1 → 4) Change cube counterclockwise (4 → 1)

INT FLUO ↑ INT FLUO ↓ FD FLUO ↑ FD FLUO ↓ CHG FW IFW ExMan SHUTTER FL

Increase intensity (fluorescence) Reduce intensity (fluorescence) Open field diaphragm (fluorescence) Close field diaphragm (fluorescence) Toggle filter functions Activate external filter wheel Activate Excitation Manager Open/close fluoshutter

COMBI CHANGE COMBI

Combination method (PH fluorescence or ICT fluorescence) Cycle through all combination methods

CHANGE OBJ CW CHANGE OBJ CCW Z FINE Z COARSE XY PRECISE XY FAST BTP ON/OFF DRY/IMM CHANGE FLT CHANGE CS OBJ 1-6 MEM 1-6

Cycle through objectives clockwise Cycle through objectives counterclockwise Activate fine focusing (Leica DMI6000 B only) Activate coarse focusing (Leica DMI6000 B only) Activate precise stage Activate fast stage Bottom port on/off (Leica DMI6000 B only) Switch dry/immersion Switch TL filter Switch to confocal application Select objective at position 1-6 Memory activated stored functions

AFC ON/OFF AFC HOLD

Turns AFC on or off Holds current position

7. Start-up 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 instrument 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.

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 transported and assembled. Before performing the required steps, please familiarize yourself with the LeicaScreen and the controls.

Caution! After turning on the gas discharge lamp, the burner must be immediately adjusted. Therefore, do not turn on the power supply unit yet. First, work in transmitted light in order to familiarize yourself with the microscope’s controls.

After the initialization (Fig. 81) is complete, the LeicaScreen will display the microscope’s current settings (Fig. 82). Fig. 81 LeicaScreen Initialization

Fig. 80 front side Leica CTR6000 1 On/Off switch 2 Signal lamp

Fig. 82 LeicaScreen after Initialization

2 1

7. Start-up 7.3 The LeicaDisplay (Leica DMI 4000 B and DMI 6000 B)

Pictograms

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.

Contrast method Objective/ Magnification

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)

Illumination Diaphragm Ports/Eyepiece Focus/stops (DMI6000 B only)

The content of the display changes according to the active function.

Fig. 83 Arrangement of the function buttons – overview 1 Four variable function buttons 2 Illumination Manager 3 Front control panel 4 Focus buttons (DMI6000 B only) 5 Three variable function buttons 6 SmartMove knobs 7 SmartMove function buttons

6 1

7. Start-up The motorized DMI stands can be controlled using the function buttons on the stand, the remote control SmartMove or the STP6000. 7.4 The Function Buttons on the Stand

Note: Changes to the light intensity as well as aperture and field diaphragm settings are stored for the individual objectives and contrast methods.

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 mirrored 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 function 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 incident-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.

Fig. 84 Fixed function buttons (left side of stand) 1 Toggle transmitted light/incident light 2 Aperture diaphragm 3 light intensity 4 field diaphragm

3 2

7. Start-up Variable Function Buttons on the Stand The variable function buttons are assigned functions 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.

Fig. 85 Function buttons (left side of stand) 1 Variable function buttons 2 Open/close aperture diaphragm 3 TL/IL switching 4 Open/close field diaphragm 5 Increase/decrease light intensity

Note: The Leica Application Suite (LAS) software is required for changing the button assignments.

Possible functions*: 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 ↓ 1 2 3 45 FD FLUO ↑ FD FLUO ↓ CHG FW IFW ExMan Fig. 86 Function buttons (right side of stand) COMBI 1 Variable function buttons 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

* See page 63 for abbreviations

7. Start-up Function Buttons on the Front Panel (Fig. 87)

CUBE

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 (Leica DMI6000 B only), switching to top port: manually.

SHUTTER Opens and closes the shutter (87.3). Switches

between

the

The CUBE 1 to CUBE 6 (87.6) buttons permit the direct selection of individual filter cubes, provided the selected cube is valid for the selected method. Press the CUBE 3 and CUBE 4 buttons 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) Z↑ Z↓

Moves the Z drive in the indicated direction.

SET + Z↑

Sets the upper focus stop.

possible

→ magnifications of the magnification

→

changer (87.4). SET + Z↓ Sets the lower stop. The magnification changer is set to the magnification 1x (87.5).

Fig. 87 Front control panel 1 100% light to eyepiece 2 Toggle ports 3 Shutter 4 Switch between subsequent magnifications 5 Subsequent magnification 1x 6 Selecting filter cubes 1

Fig. 88 1 Focus control buttons 2 Open filter drawer

5 2

6 1

7. Start-up 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 drawer 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).

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 magnification (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.

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 functions 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.

• Close the field diaphragm manually until the edge of the diaphragm appears in the field of view. Fig. 89 SmartMove remote control module 1 travel in x 2 Travel in y 3 Focus 4 Individual adjustment of button height 5 Variable function buttons (factory preset)

Note: The Leica Application Suite (LAS) software is required for changing the button assignments.

4 2

5 69

8. Operation 8.2 Contrast Methods

8.2.1

Bright Field (TL)

All of the contrast methods of the Leica DMI4000 B and Leica DMI6000 B can be selected and controlled via the variable function buttons and the Leica Application Suite (LAS). The only exceptions are methods that involve components requiring manual control (e.g. systems with manual analyzers). The following section describes the use of the function buttons on the stand. For instructions on the use of the software, please refer to the separate manual.

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.

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.

• 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, filter cube „A“ can be swapped for filter cube „ATL“ using the Leica Application Suite (LAS/LAS AF). TL contrast methods are possible with that filter cube.

8. Operation • Use the TL/IL function button to switch to transmitted light (TL).

• Insert a transmitted light specimen. • Rotate an appropriate objective into place.

• 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) 1 variable function buttons 2 Open/close aperture diaphragm 3 TL/IL switching 4 Open/close field diaphragm 5 Increase/decrease light intensity

2 3 4 5

• Focus the image with the knob on the SmartMove or the focusing wheel and adjust the intensity with the INT function buttons.

Fig. 109 Function buttons (right side of stand) 1 variable function buttons

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 completely. • 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 method 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 SmartMove or the focusing wheel and adjust the intensity with the INT function buttons.

Note: When selecting the phase contrast method, the aperture diaphragm is opened fully and can not be adjusted.

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).

Leica DMI4000 B and Leica DMI6000 B: • Use the TL/IL function button to switch to fluorescence 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 LeicaScreen:

• The filter cubes are swiveled manually into the beam path by turning the incident light turret disk. 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 SmartMove or the focusing wheel and adjust the intensity with the INT function buttons.

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