DM4000B & M, DM4500P and DM500B Operating Instructions Sept 2007

Copyrights

Copyrights All rights to this documentation are held by Leica Microsystems CMS 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 CMS 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.

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Contents

Contents 1.

Important notes about this manual ...

7

2.

Intended purpose of the microscope ...

8

3. 3.1 3.2 3.3

Safety notes ... 9 General safety notes ... 9 Electrical safety ... 10 Disposal ... 11

4.

Overview of the instrument ... 12

5.

Unpacking the microscope ... 17

7. 7.1 7.2 7.3 7.4 7.5

7.6. 7.7 7.8

6. 6.1 6.2 6.3 6.4 6.5

Assembling the microscope ... Specimen stage ... Condenser ... Tube and eyepieces ... Objectives ... Light sources for the transmitted light axis ... 6.6 Light sources for the incident light axis ... 6.7 Equipping the incident light turret disk ... 6.8 Polarizer and analyzer ... 6.9 DIC prisms ... 6.10 Optional accessories ... 6.11 Connection to the power supply ... 6.12 Connecting to the CTR5000 electronics box ...

19 19 21 22 23 23 25 30 31 32 33 34

Startup ... Functional principle ... Switching on the unit ... The display (DM4000 B/4500 B/4000 M/4500 P) ... The function keys ... Köhler illumination ... 7.5.1 Transmitted light ... 7.5.2 Incident light ... Checking the phase contrast rings ... Setting the motorized polarizer (DM4500 P/DM5000 B) ... Adjusting the light sources ...

35 35 38 39 40 41 41 42 44 45 45

8. Operation ... 51 8.1 Switching on the unit ... 51 8.2 Stages and object displacement ... 51 8.3 Focusing ... 53 8.4 Tubes ... 53 8.5 Eyepieces ... 55 8.6 Objectives ... 55 8.7 Magnification changer ... 58 8.8 HC P 1x/1.6x tube optics ... 58 8.9 Light sources ... 59 8.10 Aperture diaphragm and field diaphragm ... 59

34

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Contents 9.

Contrast methods for Leica DM4000 B/DM4500 B/ DM4500 P/DM5000 B ... 9.1 Transmitted light ... 9.1.1 Bright field ... 9.1.2 Phase contrast ... 9.1.3 Dark field ... 9.1.4 Polarization ... 9.1.4.1 Manual method ... 9.1.4.2 DM4500 P - examinations in polarized transmitted light ... 9.1.4.3 Motorized method ... 9.1.4.4 Combined methods ... 9.1.5 Differential interference contrast ... 9.1.5.1 DM4500 B/DM4500 P ... 9.1.5.2 DM5000 B ... 9.2 Fluorescence ...

68 68 69 70

10. Contrast methods for Leica DM4000 M ... 10.1 Incident light ... 10.1.1 Bright field ... 10.1.2 Dark field ... 10.1.3 Polarization ... 10.1.4 Interference contrast ... 10.2 Transmitted light ... 10.2.1 Bright field ... 10.2.2 Polarization ...

71 71 71 71 72 73 73 73 73

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11. Troubleshooting ... 74 60 60 60 60 61 61 61

12. Care of the microscope ... 12.1 Dust cover ... 12.2 Cleaning ... 12.3 Handling acids and bases ...

77 77 77 78

13. Essential wear and spare parts ... 79 14. Abbreviations and pictograms ... 80

62 68 68

15. Index ... 81 16. EU Declaration of Conformity ... 82

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 and put into operation.

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.

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.

Instructions on disposing of the microscope, accessory components and consumables. Item not contained in all configurations. * 7

2. Intended purpose of the microscope

2. Intended purpose of the microscope The DM4000 – DM5000 microscopes to which these operating instructions belong, and which have the identifying letter B, are intended for biological routine and research applications. This includes examining specimens taken from the human body for the purpose of gaining information about physiological or pathological conditions or inborn anomalies, or testing for safety and compatibility for potential recipients, or for monitoring therapeutic measures. The microscopes that have the identifying letters M or P are intended for materials science, geological or mineralogical examinations. The above-named microscopes comply with the Council Directive 98/79/EEC concerning in vitro diagnostics. They also conform to the Council Directives 73/23/EEC concerning electrical apparatus and 89/336/EEC concerning electromagnetic compatibility for use in an industrial environment.

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Caution! The manufacturer assumes no liability for damage caused by, or any risks arising from using the microscopes for other purposes than those for which they are intended or not using them within the specifications of Leica Microsystems CMS GmbH. In such cases the declaration of conformity shall cease to be valid.

Caution! These (IVD) instruments are not intended for use in the patient environment defined by DIN VDE 0100-710. Nor are they designed to be combined with medical instruments in accordance with EN 60601-1. If a microscope is electrically connected to a medical instrument in accordance with 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 was built and tested in accordance with the safety regulations for electrical measuring, control, regulating and laboratory devices in accordance with EN 61010-2-101:2002 EN 61010-1:2001 IEC 1010-1:2001

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.

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, Germany, 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. Unauthorized alterations to the device or noncompliant use shall void all rights to any warranty claims and void product liability!

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3. Safety notes 3.2 Electrical safety

ebq 100 supply unit*

General specifications

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

Leica CTR5000 electronics box (for DM5000 B) For indoor use only. Supply voltage: Frequency: Power input: Fuses: Ambient temperature: Relative humidity: Overvoltage category: Pollution degree:

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

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

Caution! Microscope For indoor use only. Supply voltage: Frequency: Power input: DM4000 DM4500 DM5000 Fuses: DM4000 DM4500 DM5000 Ambient temperature: Relative humidity: Overvoltage category: Pollution degree:

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90-250 V~ 50-60 Hz max. 180 VA max. 180 VA max. 290VA T6.3 A (IEC 60127-2/3) T6.3 A (IEC 60127-2/3) See CTR5000 15-35°C max. 80% to 30°C II 2

The power plug 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!

Caution! Through connection to the grounding connection, ancillary equipment with its own and/or extra power supply may be brought to the same ground wire potential. For connections without a ground connector, Leica Service must be consulted.

3. Safety notes 3.3 Disposal Caution! Never use any fuses as replacements other than those of the types and the current ratings listed here. Bypassing fuse holders is not permitted.

Once the product has reached the end of its service life, please contact Leica Service or Sales about disposal. Please observe and comply with the national and federal laws and regulations that are equivalent to EU guidelines such as WEEE.

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

Like all electronic devices, the microscope, its accessory components and consumables must never be disposed of with general household waste.

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. Operating 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.

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4. Overview of the instrument

4. Overview of the instrument Specification

Contrast methods

Transmitted light axis

Incident light axis

Z pinion Objective turret

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Leica DM4000 B

Leica DM4000 M

Leica DM5000 B

Leica DM4500 P

• Transmitted light: DM4000 B: BF, DF, PH, Pol DM5000 B: and ICT (mot.) • Incident light: Fluorescent

• Transmitted light: DM4000 M: BF, DF, PH, ICT, Pol DM4500 P: BF, DF, PH, ICT, Pol (conoscopy) • Incident light: BF, DF, ICR, Pol, Fluo

• Automatic illumination manager (mot. aperture diaphragm and field diaphragm, mot. intensity control) • Automatic constant-color intensity control • Motorized shutter • Integrated into the stand • Motorized 5x filter turret disk (DM5000 B 8x optional) • With FIM (fluorescence intensity manager) for reducing the light intensity in 5 increments • Mechanical booster lens for increasing fluorescence intensity • Motorized shutter

• Integrated into the stand • Motorized 4x filter turret disk • Automatic illumination manager • DM4000 M: motorized shutter

• Manual • Manual, fully encoded • DM4000 B: 6x/7x with M25 thread DM5000 B: 7x (M25) DM5000 B: With object prism disk (3 positions)

• Manual, fully encoded • DM4000 M: 6x with M32 thread DM4500 P: 6x with M25 thread, centerable, encoded • Receptacle for DIC prisms and Pol compensators (for DM4000 M: optional)

4. Overview of the instrument

Specification

X/Y stage

Leica DM4000 B

Leica DM4000 M

Leica DM5000 B

Leica DM4500 P

• Manual • Replaceable specimen stage • Coaxial drive length: 155 mm

• Manual • DM4000 M: • Replaceable specimen stage • Coaxial drive length: 140 mm DM4500 P: • Replaceable Pol stage

Tube

• Manual or motorized (DM4500P: manual) • Optionally with one or two camera outputs • DM4500 P: conoscopy module (tube optics HC P1x/1.6x with Bertrand lens, encoded)

Condenser

• Motorized condenser head • Condenser disk for the light ring, DF-Stop, DIC prisms • Automatic Köhler illumination • Optional polarizer (integrated and motorized)

Magnification changer (optional)

• Manual • 3x fully encoded • 1x; 1.25x; 1.6x

• Manual • 3x fully encoded • 1x; 1.5x; 2x

Controls

• Operating buttons on the stand for all motorized microscope functions • Additional variable multifunction keys • Focus wheels • LCD • DM5000 B with Leica SmartTouch

Computer interface

• USB2.0

Software tools

• Leica Application Suite (LAS) for WindowsTM 2000, XP, Vista • With plug-ins for: • Microscope and camera configuration • Microscope and camera control • Image acquisition 13

4. Overview of the instrument Specification

Electronics box Leica CTR5000

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Leica DM4000 B

Leica DM4000 M

Leica DM5000 B

Leica DM4500 P

Only for the Leica DM5000 B: Separate operating unit with a power supply for 100 W halogen lamps. See → p.10 (Electrical safety)

4. Overview of the instrument

1 2

3

14 4

5 6

7 13

Fig. 1 1 2 3 4 5 6 7

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11 10 9 8

Left side of the stand with the advanced AET22 ErgoTube

Eyepiece Eyepiece tube Tube Objective turret with objectives Specimen stage with specimen holder Condenser LCD

8 Field diaphragm operating buttons 9 Transmitted light / incident light switch 10 Aperture diaphragm operating buttons 11 Brightness adjustment buttons 12 Focus wheel with coarse and fine adjustment 13 Variable function keys (preset at the factory) 14 Lamp adjustment window

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4. Overview of the instrument

15 22

16

21

Fig. 2

Right side of the stand with the advanced ErgoTube AET22

15 Lamp housing for incident light 16 Lamp housing for transmitted light 17 Transmitted light filter, optional 18 Transmitted light filter, optional 19 Variable function keys (preset at the factory) 20 x/y coaxial drive, adjustable height 21 Handwheel for fine focus 22 Motorized filter block exchanger

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20

19 18

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5. Unpacking the microscope

5. Unpacking the microscope The microscope is delivered in two packages. The stand package contains the following components: • Stand with integrated incident light axis and objective turret • Specimen stage with stage bracket

The external ebq 100 supply unit* is delivered in a separate package. For the Leica DM5000 B microscope: The Leica CTR5000 electronics box is delivered in a separate package. First, carefully remove all components from the transportation and packaging materials.

• Power cable and PC connecting cable • CD with Leica Application Suite (LAS) software package • Instructions and list of microscope default settings The system package contains the microscope’s accessories:

Note: If at all possible, avoid touching the lens surfaces of the objectives. If fingerprints do appear on the glass surfaces, remove them with a soft leather or linen cloth. Even small traces of finger perspiration can damage the surfaces of optical devices in a short time. See the chapter on "Care of the microscope" → p. 77 for additional instructions.

• Tube • Eyepieces • Objectives • Condenser

Caution! Do not connect the microscope or peripherals to an AC power source at this time under any circumstances!

• Lamp housings with accessories • Assembly tools • Additional microscope accessories such as filter cubes, etc. depending on product configuration

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5. Unpacking the microscope Installation location

Transport

Work with the microscope should be performed in a dust-free room, which is free of oil vapors and other chemical vapors, as well as extreme humidity. At the workstation, large temperature fluctuations, direct sunlight and vibrations should be avoided. These conditions can distort measurements and micrographic images.

For shipping or transporting the microscope and its accessory components, the original packaging should be used.

Allowable ambient conditions Temperature 15-35°C Relative humidity maximum 80% up to 30°C

• Unscrew the objectives.

Microscopes in warm and warm-damp climatic zones require special care in order to prevent the build up of fungus. See the chapter on "Care of the microscope" → p. 77 for additional instructions.

• Remove the specimen stage.

As a precaution to prevent damage from vibrations, the following components should be disassembled and packaged separately:

• Remove the condenser.

• Remove the lamp housings. • Disassemble the burner of 106 z lamp housing. • Remove all moving or loose parts.

Caution! Electrical components must be assembled at least 10 cm away from the wall and from flammable substances.

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6. Assembly

6. Assembling the microscope The microscope components are logically assembled in this order: • Specimen stage • Condenser with condenser head • Tube • Eyepieces • Objectives • Lamp housings with light sources • Equipment for the incident light turret disk* Only a few commonly used screwdrivers and keys are necessary for assembly; these are included in the delivery package. When using intermediate systems and optical accessories, the sequence may vary. In this case, read chapter, "6.10 Optional accessories" → p.33

6.1 Specimen stage

!

Caution:

Never install objectives before assembling the stage. • Place the specimen holder on the stage and fasten it with the two screws (3.1). • Using the condenser height adjuster (3.2), turn the condenser holder completely upwards, i.e. as close to the stage as possible. • Loosen the stage clamp (3.3) slightly.

Fig. 3 Mechanical object stage 1 Locking screws for specimen holder 2 Condenser height adjuster 3 Stage clamp

1

2

3

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6. Assembly • From above, set the stage clamp onto the dovetail guide (4.2) and push the stage downwards until the upper end of the dovetail guide is tightly fastened to the upper end of the stage clamp. • Firmly tighten the stage clamp (4.1).

Note: For thicker specimens (Leica DM4000 M) the stage can be set to a correspondingly lower level.

• Only for DM4500 P: Pol attachable mechanical stage* Adjust the attachable mechanical stage so that the fastening screw is visible below the holes (4a.1). Set the attachable mechanical stage in the guide holes on the rotating stage and tighten the fastening screw using the hexagonal key. Attachable mechanical stage* The attachable mechanical stage can be installed on the left, on the right or on the front (not pictured). The two clamping screws fasten it into place.

Fig. 4a Pol rotating stage* and Pol 3 attachable mechanical stage* 1 Holes for the fastening screw. 2 Lever for the holder for glass slides of various formats, which can be turned inward and outward 3 Storage for the centering key 4 Locking button pair 5 45° click stop 6 Clamping system for the stage rotation

Fig. 4 Assembling the stage 1 Stage clamp 2 Dovetail guide

4 3

1 1

2

5

2 6

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6. Assembly 6.2 Condenser • Screw the condenser head into the condenser. • Using the condenser height adjuster (5.4), turn the condenser holder (5.1) downward as far as it will go. • Unscrew the clamping screw for the condenser (5.3) far enough so that the condenser can be inserted from the front.

Note: The condenser must be centered before using the microscope. → Köhler illumination p. 41. Fig. 6 Underside of condenser 1 Orientation pin

1

• From the front, insert the condenser into the condenser holder as far as it will go. On the underside of the condenser, there is an orientation pin (6.1) that must be locked into place in the guiding notch (7.1). • Tighten the condenser’s clamping screw (5.3) until the condenser locks into place.

Fig. 7 Condenser holder 1 Guiding groove

• Connect the condenser over the connection (8.1) with the stand. 1

Fig. 5 Condenser holder 1 Condenser holder 2 Condenser centering 3 Clamping screw for the condenser 4 Condenser height adjuster

Fig. 8 Condenser connection 1 Condenser cable socket

1

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4

1 21

6. Assembly 6.3 Tube and eyepieces The tube is mounted on the stand either directly or with the use of intermediate modules. The side clamping screw fastens it into place (9b.1). • For the MBDT motorized tube only: Remove the transport anchor (9a.1) on the underside of the tube.

• The eyepieces are inserted into the eyepiece tubes on the tube. • For the BDTP tube only: The Pol eyepieces are inserted into the eyepiece tubes (using the locking groove).

• Partially unscrew the clamping screw (9b.1). • Insert the tube into the circular receptacle (dovetail ring). • Retighten the clamping screw (9b.1).

Fig. 9b Fastening the tube 1 Clamping screw

• For the MBDT motorized tube only: Connect the tube to the stand with the connector bushing (10.1).

Fig. 9a Underside of the tube 1 Transport anchor

1

Fig. 10 Motorized tube connection 1 Connector socket

1

1

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