Leica Microsystems

DMI6000 B Operating Manual Oct 2004

Operating Manual

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Leica DMI6000 B Operating Manual

Published October 2004 by:

Leica Microsystems Wetzlar GmbH Ernst-Leitz-Straße D-35578 Wetzlar (Germany)

Responsible for contents: Bernard Kleine (Marketing CM, Compound Microscopy, Product Management) Holger Grasse (Safety Officer according to MPG §30)

In case of questions, please contact the hotline: Phone

+49(0)6441-292286

Fax

+49(0)6441-292255

E-mail: [email protected]

Leica DMI6000 B Operating Manual

Copyrights

Copyrights All rights to this documentation are held by Leica Microsystems Wetzlar GmbH. Reproduction of text or illustrations (in whole or in part) by print, photocopy, microfilm or other method (including electronic systems) is not allowed without express written permission from Leica Microsystems Wetzlar GmbH. The term "Windows" may appear in the following text without further identification. It is, however, a registered trademark of Microsoft Corporation. The names of companies and products used herein may be trademarks of their respective owners. The instructions contained in the following documentation reflect state-of-the-art technology and knowledge standards. We have compiled the texts and illustrations as accurately as possible. Nevertheless, no liability of any kind may be assumed for the accuracy of this manual’s contents. Still, we are always grateful for comments and suggestions regarding potential mistakes within this documentation. The information in this manual is subject to modification at any time and without notification.

Contents

Contents 1.

Important Notes about this Manual ...

Intended Purpose of the Microscope ...

3. Safety Notes ... 9 3.1 General Safety Notes ... 9 3.2 Electrical Safety ... 10 4. Overview of the Leica DMI 6000 B ... 12 4.1 Specifications ... 12 4.2 Glossary ... 16 5.

Unpacking the Microscope ... 22

6. Assembling the Microscope ... 6.1 Assembly Tools ... 6.2 Installation of the Transmitted Light Illumination Carrier (DL) ... 6.3 Installation of the DIC Module and DIC Objective Prisms ... 6.4 Installation of Specimen Stages ... 6.5 Installation of Condensers ... 6.6 Installation of Eyepieces ... 6.7 Installation of Objectives ... 6.8 Installation of Filters in the Illumination Arm ... 6.9 Installation of the Transmitted Light Lamp Housing ... 6.10 Installation and Replacement of the Transmitted Light Lamps: ... Lamp Housing 107 or 107/2 ...

25 25 26 27 28 34 39 39 40

6.11 Installation of 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 Installation of the Polarizer and Analyzer ... 6.15 Optional Accessories ... 6.16 Connection to the Electronics Box CTR6000 ... 6.17 Connection to the Computer ... 6.18 Connection to the Power Supply ... 7. 7.1 7.2 7.3 7.4 7.5 7.6

7.7 7.8 7.9

Start-up ... Functional Principle ... Switching on the Microscope ... The LeicaScreen ... The Function Buttons on the Stand ... The SmartMove Remote Control Module ... Illumination ... 7.6.1 Transmitted Light ... 7.6.2 Incident Light - Fluorescence ... Checking Phase Contrast Rings ... Setting the Motorized Polarizer ... Adjusting the Light Sources ...

42 44 48 50 52 53 54 54 55 55 59 60 61 64 65 65 68 69 70 71

Contents 8. Operation ... 8.1 Switching On ... 8.2. Contrast Methods ... 8.2.1 Brightfield (TL) ... 8.2.2 Phase Contrast (TL) ... 8.2.3 Darkfield (TL) ... 8.2.4 Polarization (TL) ... 8.2.5 Differential Interference Contrast (TL) ... 8.3 Fluorescence ... 8.4 Combination Methods ... 8.5 Focusing ... 8.6 Tubes ... 8.7 Eyepieces ... 8.8 Objectives ... 8.9 Stages and Object Displacement ... 8.10 Magnification Changer ... 8.11 Light Sources ... 8.12 Aperture and Field Diaphragm ... 9.

74 74 76 76 77 77 78 79 80 81 82 84 85 85 88 89 89 90

Troubleshooting ... 91

10. Care of the Microscope ... 10.1 Dust Cover ... 10.2 Cleaning ... 10.3 Handling Acids and Bases ...

95 95 95 96

11. Major Consumable and Replacement Parts ... 97 12. Dimensions ... 98 13. Abbreviations and Pictograms ... 99 14. Index ... 101 15. EU Declaration of Conformity ... 104

1. Important Notes about this Manual

1. Important Notes about this Manual Caution! This operating manual is an essential component of the microscope, and must be read carefully before the microscope is assembled, put into operation or used.

This operating manual contains important instructions and information for the operational safety and maintenance of the microscope and accessories. It must therefore be kept safely for future reference. A separate manual is available on CD-ROM covering the operation of the Leica Application Suite (LAS).

Text symbols, pictograms and their meanings: (1.2)

Numbers in parentheses, such as "(1.2)", correspond to illustrations (in the example, Figure 1, Item 2).

→ p.20 Numbers with pointer arrows (for example → p.20), point to a certain page of this manual. Caution! Special safety instructions within this manual are indicated with the triangle symbol shown here, and have a gray background.

Caution! The microscope and accessories can be damaged when operated incorrectly.

Explanatory note.

Item not contained in all configurations.

2. Intended Purpose of the Microscope

2. Intended Purpose of the Microscope The Leica DMI 6000 B microscope described in these instructions is intended for biological routine and research applications. This includes the examination of samples taken from the human body to provide information on physiological or pathological states or congenital abnormalities, or determing the safety and compatibility with potential recipients, or monitoring therapeutic measures.

The above-named microscope complies with the Council Directive 98/79/EEC concerning in vitro diagnostics. It also conforms to the Council Directives 73/23/EEC concerning electrical apparatus and 89/336/EEC concerning electromagnetic compatibility for use in an industrial environment.

Caution! The Leica DMI 6000 B is a further development of Leica’s proven inverted research microscopes. It is designed for cellular and tissue examination, micromanipulation and microinjection techniques, microdissection, and confocal microscopy. The Leica DMI 6000 B is suitable for universal deployment, all contrast methods such as darkfield, brightfield, phase contrast, DIC, fluorescence, and modulation contrast are integral to the microscope and can be adapted or changed quickly and easily. Variable illumination and imaging beam paths, as well as HCS optics, modular accessories and a comprehensive range of peripherals, complement the Leica DMI 6000 B inverted research stand.

The manufacturer assumes no liability for damage caused by, or any risks arising from using the microscope for other purposes than those for which it is intended or not using it within the specifications of Leica Microsystems Wetzlar GmbH. In such cases the declaration of conformity shall cease to be valid.

Caution! This (IVD) device is not intended for use in the patient environment defined by DIN VDE 0100-710. Neither is it intended for combining with medical instruments according to EN 60601-1. If a microscope is electrically connected to a medical instrument according to EN 60601-1, the requirements defined in EN 60601-1-1 shall apply.

3. Safety Notes

3. Safety Notes 3.1 General Safety Notes This safety class 1 device is constructed and tested in accordance with EN 61010-2-101:2002, EN 61010-1:2001, IEC 1010-1:2001, Safety regulations for electrical measuring, control, and laboratory devices.

Caution! The devices and accessories described in this operating manual have been tested for safety and potential hazards. The responsible Leica affiliate or the main plant in Wetzlar must be consulted whenever the device is altered, modified, or used in conjunction with non-Leica components that are outside of the scope of this manual.

Caution! In order to maintain this condition and to ensure safe operation, the user must follow the instructions and warnings contained in this operating manual.

Unauthorized alterations to the device or noncompliant use shall void all rights to any warranty claims!

3. Safety Notes 3.2 Electrical Safety Caution!

General specifications Leica CTR6000 Electronics Box For indoor use only. Supply voltage: Frequency: Power input: Fuses: Ambient temperature: Relative humidity: Overvoltage category: Pollution degree:

90-250V~ 50-60 Hz max. 290VA T6.3 A (IEC 60127-2/3) 15-35°C max. 80% to 30°C II 2

Power plugs may only be plugged into an outlet equipped with a grounding contact. Do not interfere with the grounding function by using an extension cord without a ground wire. Any interruption of the ground wire inside or outside of the device, or release of the ground wire connection, can cause the device to become hazardous. Intentional ground interruption is not permitted!

Microscope For indoor use only. Supply voltage: Frequency: Power input: Fuses: Ambient temperature: Relative humidity: Overvoltage category: Pollution degree:

Caution! 90-250V~ 50-60 Hz See CTR6000 See CTR6000 15-35°C max. 80% to 30°C II 2

Peripheral devices with their own or separate power supplies that are connected to the microscope can have the same protective conductor potential by connecting them to the ground screw on the back of the Leica CTR6000 electronics box. For connections without a ground connector, Leica Service must be consulted.

ebq 100 Supply Unit* For indoor use only. Supply voltage: Frequency: Power input: Fuses: Ambient temperature: Relative humidity: Overvoltage category: Pollution degree: (See enclosed manual)

90-250V~ 50-60 Hz max. 155VA 2xT2A (IEC 127) 10-36°C max. 80% to 30°C II 2

Caution! Never use any fuses as replacements other than those of the types and the current ratings listed here. Using patched fuses or bridging the fuse holder is not permitted. The use of incorrect fuses may result in a fire hazard.

3. Safety Notes

Caution! The microscope’s electrical accessory components are not protected against water. Water can cause electric shock.

Caution! Protect the microscope from excessive temperature fluctuations. Such fluctuations can lead to the accumulation of condensation, which can damage the electrical and optical components. Ambient temperature: 15-35°C.

Caution! Before exchanging the fuses or lamps, be absolutely certain to switch off the main power switch and remove the power cable.

4. Overview of the Instrument

4. Overview of the Leica DMI 6000 B 4.1 Specifications Contrast Methods

• Transmitted light (DL): • Incident light (IL): • Combination (DL/IL): • Intermediate pupil:

Transmitted Light Axis

• Automatic Illumination Manager (aperture, field diaphragm, intensity, process switching) • Automatic, color-neutral intensity control • Manual or motorized shutter • Lamp housing mount for interchangeable lamp housings. • Automatic, electronic condenser identification

Incident Light Axis

• Aautomatic Illumination Manager (aperture, field diaphragm, intensity, process switching) • Automatic, color-neutral intensity control • Motorized shutter (switching speed < 50ms) • Lamp housing mount for up to 3 interchangeable light sources • Motorized 6-place filter turret • Fluorescence Intensity Manager (FIM) (reduction of incident illumination intensity) • Mechanical booster lens for central boosting of fluorescence or uniform distribution • Motorized Excitation Manager to monitor fluorescence emission when using double and triple filter cubes • Ultrafast filter wheel for 3 excitation wavelengths (switching speed < 50 ms)

Tube

BF, DF, PH, DIC, Pol Fluo Fluo/DIC, Fluo/PH IMC (integrated modulation contrast) IPH (integrated pos./neg. phase contrast)

• Ergonomic with or without camera port at left • 2 switching positions: 100%VIS and 50%VIS / 50%CAM • Optional Bertrand lens • Eye spacing adjustment • Height and angle adjustment (10° - 45°)

4. Overview of the Instrument Magnification Changer

• Motorized • 3 switching positions (choice of magnifications: 1x, 1.5x, 1.6x or 2.0x) • Effective for all camera ports or • Manual • 2 switching positions (choice of magnifications: 1x, 1.5x, 1.6x or 2.0x) • Effective on tube port

Objective Turret

• Motorized and coded • 6x for objectives with M25 thread and 45mm parfocal distance • For DIC: motorized/coded Wollaston prism carousel • Anti-vibration locking

Stages

Fixed regular stages • Ceramic-coated stage plate (248mm x 204mm) • Heated stage plate (from 3°C above room temperature to 60°C) (248 x 212mm) • Temperature-controlled stage plate (0°C to 60°C) (248mm x 212mm) Fixed micromanipulation stages • Ceramic-coated stage plate (248mm x 204/122mm) • Heated stage plate (from 3°C above room temperature to 60°C) (248mm x 204/122mm) • Temperature-controlled stage plate (0°C to 60°C) (248mm x 204/122mm) Regular manual 3-plate cross-stage • Positioning range: 83mm x 127mm • 20 optional inserts (standard, heating, cooling) for a variety of applications, size of inserts:160mm x 110mm (compatible with scanning stages) Manual micromanipulation 3-plate cross-stage • Positioning range: 40mm x 40mm • 3 optional inserts for a variety of applications Motorized micromanipulation 3-plate cross-stage • Positioning range: 40mm x 40mm Scanning stage IM 120 x 100 (motors on top) • 1mm, 2mm, 4mm spindle pitch (higher resolution v. higher speed) • 20 optional inserts (standard, heating, cooling) for a variety of applications, size of inserts:160mm x 110mm 13

4. Overview of the Instrument

Scanning stage IM 120 x 100 (motors on bottom) • 1mm, 2mm, 4mm spindle pitch (higher resolution vs. higher speed) • 20 optional inserts (standard, heating, cooling) for a variety of applications, size of inserts:160mm x 110mm Condensers

• Motorized and coded • Motorized or manual aperture diaphragm • Contrast methods: BF, DF, PH, DIC, Pol, IMC • Automatic method switching • Condenser turret with 7 positions for contrast methods • 2 condenser housings (S1-S28 and S70) • Condenser heads: S1/1.4 oil, S1/0.9 dry, S23/0.53, S28/0.55 • Condenser heads can be swung out • Condenser S70 with additional lens for low magnifications • All condensers suitable for magnifications from 1.25x to 100x • With or without motorized or manual polarizer • With or without motorized or coded Wollaston prism disk

Z focus

• Motorized and coded • 9mm travel (1mm below, 8mm above the stage) • Maximum travel speed: 5mm/s • 5 focus steps: 0.05 µm, 0.1 µm, 0.7 µm, 1.5 µm, 5.0 µm • Electronic focus repositioning • Automatic lowering prior to objective change • Electronic parfocality

Observation Ports

• Motorized and coded • Left side ports (100%, 80%, or 50% transmission) • Left side port dichroic splitting at 680 nm • Right side ports (100%, 80%, or 50% transmission) • Bottom port Optional • Top port with 2 switching positions • 100% to eyepieces • 50% to eyepieces/ 50% to port

4. Overview of the Instrument

Controls

• 7 fixed control buttons for illumination and apertures • 7 variable function buttons behind the focus controls • 3 fixed control buttons for focus steps • 2 focus hand wheels • 7 buttons for fluorescence cubes and shutters • 4 buttons for magnification changers and ports • SmartMove: ergonomic controller for x, y, z and 4 additional variable function buttons

Electronics Box Leica CTR6000

• Separate control unit for all motorized and electronic elements of the microscope such as: • Objective turret • Ffocus • Ports • Magnification changer • Fluorescence • Condenser • Motorized stages With • Power supply for 100W halogen lamp • Power supply for SmartMove

Interfaces

• 2 x RS232C • 2 x USB • 4 x external/internal peripherals

Software Tools

• Leica Application Suite (LAS) for WindowsTM 2000, XP with plug-ins For: • Microscope and camera configuration • Microscope and camera control • Image acquisition

4. Overview of the Instrument 4.2 Glossary

Eyepieces

The Stand

The eyepieces create an enlarged, virtual image of the actual intermediate image created by the objective. The eyepiece serves as a magnifier in this respect.

Four basic versions of the Leica DMI stand (DMI 3000 B, DMI 4000 B, DMI 5000 B, DMI 6000 B) are available which can be combined into a wide range of microscope variants. The basic building blocks of the Leica DMI6000 B stand are: • DMI6000 B electronic stand • Integrated fluorescence axis with motorized filter cube changer (6 positions) • With or without bottom port • Lateral camera ports, 100%, 80%, or 50% • Optional top port at the left side of the tube • Integrated motorized tube lens changer • Optional Bertrand lens The individual variants and their components, differences, and applications are described in this manual. The function and performance of all microscopy techniques and required accessories of the Leica DMI6000 B will be described in detail in the section of this manual that covers the operation of the microscope.

Intensity Controller The stand contains a 12V 100 W transformer for continuous regulation of the intensity via the intensity controller. The intensity can be adjusted using the controls (1.6). Focus Wheel The focus wheel allows quick, precise focusing of the microscopic image. Focusing is realized by the vertical travel of the objective turret, with a total range of 9mm. Incident Light-Fluorescence Unit The stand features an integrated fluorescence axis and a motorized filter cube changer with 6 positions. Aperture Ddiaphragm

Tube The tube and tube lens create the primary image together with the objective. The tubes are an integral part of the stand and consist of a basic body and a binocular section. The trinocular tube also features a photo/video port. A switchable mirror diverts either 100% of the light to the eyepieces or camera port, or splits it, with 50% each going to the eyepieces and camera port. A Bertrand lens is also available as an option.

The aperture diaphragm determines the resolution, depth of field, and contrast of the microscopic image. The best resolution is obtained when the apertures of the objective and the condenser are roughly the same. Caution: The aperture diaphragm in the illumination light path is not for setting the image brightness. This should be done only with the intensity controls or neutral filters.

4. Overview of the Instrument Condenser The condenser is a lens system that gathers light and projects it onto the specimen from above. The condenser is designed for the utilization of the numerical aperture in the objective. Condenser Height Adjustment The markings of the transmitted-light column indicate the height to be set for the used condenser. Stages and Accessories The stage is designed to accommodate the specimens to be observed. Mechanical and motorized 3-plate cross-stages are available for the Leica DMI 6000 B. Motorized Objective Turret and Objectives The motorized objective turret is designed to accommodate the objectives. The L-objectives with their long working distances especially take into consideration the correction of varying container bottom thicknesses. All microscope objectives are usable, from 1.25:1 to 100 : 1 magnification. All objectives in the Leica product range with a 25mm thread and coverslip correction are compatible. For performance data on Leica objectives, please refer to the most current valid objective lists available from your Leica representative. Transmitted Light Illumination Unit The transmitted light illumination unit consists of an illumination carrier and the transmitted light illumination column. The transmitted light illumination carrier does not contain a lamp housing, but a filter module for two swing-in filters and, depending on the condenser used, a field diaphragm. Filter

The filters are generally used to enhance the contrast of the specimen and are installed in the illumination carrier. A selection of various filters can be interchanged as required. Field Diaphragm The field diaphragm is used to realize Koehler illumination. Lamp Housing for Transmitted Light The lamp housings 107/2 and 107 (both for 12V 100W halogen) are available for the Leica DMI 6000 B. For their descriptions and applications, please refer to the section of this manual describing the use of the microscope. The letter L indicates a lamp housing designed for left-handed operation. Lamp Housing for Incident Light The 106 z L lamp housing (for halogen or xenon) is available for the Leica DMI 6000 B. For their descriptions and applications, please refer to the section of this manual describing the use of the microscope. The letter L indicates a lamp housing designed for left-handed operation. Leica CTR6000 Electronics Box The Leica CTR6000 electronics box contains the power supply for the lamp and the circuit boards required to control the motorized functions of the stand.

4. Overview of the Instrument

1 16 2 15 3

Fig. 1 Leica DMI 6000 B left view 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

9 8 7

11 Variable function buttons 12 Right side port 13 Booster lens 14 Lamp mount 15 Condenser head 16 Condenser base 17 Field diaphragm 18 Transmitted light lamp housing 19 DIC objective prism disk

4. Overview of the Instrument

4 5

Fig. 2 Leica DMI 6000 B right view 1 E-focus control buttons 2 Focus wheel 3 Variable function buttons 4 Opener for drawer 5 Drawer 6 Right side port

7 8

7 Analyzer slot 8 Centering window 9 Field diaphragm centering 10 Incident light lamp housing 11 Objective turret 12 Stage with attachable mechanical stage

4. Overview of the Instrument

6 3

1 Fig. 3 Leica DMI 6000 B front view 1 LeicaScreen 2 Front control panel 3 Port switching 4 Top port 5 Manual transmitted light filters 6 Field diaphragm centering

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