Leica DM4000 B Leica DM4000 M Leica DM5000 B Operating Manual 1
Published 2003 by: Leica Microsystems Wetzlar GmbH Ernst-Leitz-Straße D-35578 Wetzlar (Germany) Responsible for contents: Katja Peter, Karin Schwab Marketing CM, Compound Microscopy, Product Management In case of questions, please contact: 2 Phone +49(0)6441-292261 Fax +49(0)6441-292255 E-mail: MQM-Hotline@leica-microsystems.
Leica DM4000 B Leica DM4000 M Leica DM5000 B Operating Manual 3
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 methods (including electronic systems) is not allowed without express written permission from Leica Microsystems Wetzlar GmbH. The term "Windows" can be used in the following text without further identification. It is a registered trademark of the Microsoft Corporation.
Contents Contents 1. Important Notes about this Manual ..... 7 2. Safety Notes .............................................. 2.1. General Safety Notes ............................... 2.2. Electrical Safety ........................................ 8 8 8 3. Overview of the Instrument .................... 10 4. Unpacking the Microscope .................... 14 5. 5.1 5.2 5.3 5.4 5.5 Assembling the Microscope .................. Stage ...........................................................
Contents 8.2 Fluorescence ............................................. 9. Imaging Procedure for Leica DM4000 M ....................................... 9.1 Incident Light ............................................. 9.1.1 Bright Field ...................................... 9.1.2 Dark Field ......................................... 9.1.3 Polarization ..................................... 9.1.4 Interference Contrast .................... 9.2 Transmitted Light ...................................... 9.2.
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 put into operation or used. This operating manual contains important instructions and information for the operational safety and maintenance of the microscope and accessories. Therefore, it must be kept and taken care of. Text symbols and their meanings: (1.2) Numbers in parentheses, such as "(1.
2. Safety Notes 2. Safety Notes 2.1 General Safety Notes 2.2 Electrical Safety This safety class 1 device is constructed and tested in accordance with EN 61010-1/IEC 1010-1, safety regulations for electrical measuring, control, and laboratory devices. General specifications 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.
2. Safety Notes Supply unit ebq 100 For indoor use only. Supply voltage: Frequency: Power input: Fuses: Ambient temperature: Relative humidity: Overvoltage category: Pollution degree: (see enclosed manual) Caution! 90-250 V~ 50-60 Hz max. 155 VA 2xT2A (IEC 127) 15-35°C max. 80% to 30°C II 2 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.
3. Overview of the Instrument 3.
3. Overview of the Instrument Specification Condenser Magnification Changer Leica DM4000 B / DM5000 B Leica DM4000 M • motorized condenser head • motorized condenser turret for light rings, DF stop, DIC prisms • optional polarizer integrated and motorized • automatic Köhler Illumination • manual • absolute coded • 1x; 1.25x; 1.6x • manual • absolute coded • 1x; 1.
3. Overview of the Instrument 1 2 3 14 4 5 6 7 13 Fig.
3. Overview of the Instrument 15 22 16 21 Fig.
4. Unpacking the Microscope 4. Unpacking the Microscope The device is delivered in two boxes. The stand box contains the following components: • Stand with integrated incident light axis and objective nosepiece • Specimen stage with stage bracket The external ebq 100 supply unit* is delivered in separate packaging. For the Leica DM5000 B microscope: The CTR5000 electronics box is also delivered in separate packaging. First, carefully remove all components from the transportation and packaging materials.
4. 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 workplace, 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.
5. Assembly 5. Assembling the Microscope The microscope components are logically assembled in this order: • • • • • • • Stage Condenser Tube Eyepieces Objectives Light sources Filter cubes/reflectors* Only a few commonly used screwdrivers and keys are necessary for assembly, which are included in the delivery package. 16 When using intermediate systems and optical accessories, the sequence may vary. In this case, read Chapter, "5.10 Optional accessories" → p.
5. Assembly 5.1 Stage ! Caution: Before assembling the stage, make sure no objectives are installed! • 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). • Place the specimen holder on the stage and fasten it with the two screws (3.1). • Using the condenser height adjuster (3.
5. Assembly 5.2 Condenser • Using the condenser height adjuster (5.4), turn the condenser holder (5.1) completely downwards. • 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. 37. 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.
5. Assembly 5.3 Tube and Eyepieces The tube is mounted to the stand either directly or with the use of intermediate modules. It is fastened in place with the side clamping screw (9.1). Fig. 9 Fastening the tube 1 Clamping screw 1 • Loosen the clamping screw (9.1). • Insert the tube in the circular receptacle (dovetail ring). • Retighten the clamping screw (9.1). • Only for the MBDT motorized tube: Connect the tube to the stand with the connector socket (10.1).
5. Assembly 5.5 Light Sources for the Transmitted Light Axis Caution: Fig. 12 Lamp housing 107/2 Releasing the fastening screw Be sure that the lamp housing is disconnected from the power supply. Unplug the power plug and the power supply during assembly. 107/2 Lamp Housing This lamp housing is used with a 12V 100W halogen lamp, which is already mounted. In case the lamp has to be removed: • Remove the fastener screw on the housing (Fig. 12). • Remove the housing by pulling it upwards.
5. Assembly 5.6 Light Sources for the Incident Light Axis Caution: During assembly, always unplug the power supply unit of the 106 z lamp housing from its socket. Never touch the glass parts of the burner with bare hands. This lamp housing is used with a 12V 100W halogen lamp or various gas discharge lamps. Inserting the 12V 100W halogen lamp into the 106 z lamp housing • Unscrew the fastening screws of the cover and lift up the cover (16.1). • Unscrew the fastening screws of the lamp mount (16.
5. Assembly • Insert the lamp with the dust cover straight into the socket until it stops. • Place the lamp housing in the incident light lamp housing receptacle (18.1) and fasten it with the clamping screw on the side. • Remove the dust cover. • Reinsert the lamp mount and retighten the fastening screw (16.8). • Connect the lamp housing to the power supply for incident light (symbol ) (18.4). Caution: Do not remove the lamp’s dust cover until after you have installed the lamp.
5. Assembly Inserting the gas discharge lamps (Hg and Xe) into the 106z lamp housing Hg and Xe lamps are powered by the separate ebq 100 supply unit. Read the separate instruction manual provided with this supply unit. The following gas discharge lamps may be used and require different lamp mounts (Fig.
5. Assembly • To open the 106 z lamp housing, unscrew the fastening screws on the cover. Caution: • Remove the transport anchorage (red plastic rod in place of the burner) in the lamp mount. To do so, remove the lower clamp (19.1). Pull up the cooling element (19.3) and turn it to the side. Detach the lower clamp system (19.2) and remove the transport anchorage. Hg 50 burner: After installation, the labeling must be upright. If a glass melt nipple is present (19a.
5. Assembly • Insert the lamp mount, with the burner installed, into the lamp housing and tighten it with the screws (20.8). • Put the lid down again. Plug in the contact plug as far as it goes and retighten the screws. • Place the lamp housing in the incident light lamp housing receptacle (21.1) and fasten it with the clamping screw on the side. • Connect the lamp housing to the power supply (22.1). Fig.
5. Assembly 5.7 Equipping the Incident Light filter turret The receptacles on the turret are numbered. According to your equipment, the individual filter and/or reflector cubes have already preassigned positions. A list is provided along with your shipment (“Identification Sheet”). Fig. 23 Filter cube front side Fig. 24 Filter cube back side Insert the filter and reflector cubes in the following manner: • Equip the incident light turret only when the microscope is switched off.
5. Assembly ICT/P transmitted light polarizer • Using the left clamping screw, fasten the ICT/P transmitted light polarizer to the underside of the condenser holder (Fig. 27). Fig. 27 Assembly of the ICT/P transmitted light polarizer 1 Clamping screw • Make sure that the red index point on the front of the polarizer is aligned with 0. • If necessary, insert the compensators (λ- and λ/4 plates) into the polarizer’s receptacle (Fig. 28). 1 Fig.
5. Assembly • Remove the plug cap on the left side of the stand. • Insert the polarizer into the receptacle until it latches in place (Fig. 30). Motorized analyzer • Insert the analyzer cube as described in section 5.7 "Equipping the Incident Light filter turret" → p. 26, in the corresponding position on the filter turret. See the list provided (“identification Sheet”) for the correct position. 6.9 DIC Prisms Fig. 30 Inserting the analyzer 1 The plug cap is replaced with the analyzer.
5. Assembly Ergomodule • Remove the clamping ring from the filter slide. For raising the eye level of the tube opening, the ergomodule may be used. It is fastened in place with the side clamping screw. • Insert the Manager. Booster Lens or Excitation • Push the cover back. • Insert the clamping ring. Mirror Housing • Place the mirror housing directly onto the lamp housing receptacle on the back of the stand and attach it using the side clamping screw.
5. Assembly After completing the assembly work, connect the stand to the power supply using the power cable (Fig. 34.2). 5.12 Connection to the CTR5000 Electronics Box Fig. 34 Rear side of stand Leica DM4000 B/M 1 Power switch 2 Power supply Only for the Leica DM5000 B: • Connect the microscope (36.1) to the "Microscope" jack (35.1) on the rear of the electronics box. Use the cable with the 25-pin plug. • Connect the electronics box to the power supply using the power cable (35.2). 1 2 Fig.
6. Startup 6. Startup 6.1 Functional Principle The microscope’s most important functions may be easily accessed using function keys. • The microscope may be switched between various contrast processes by pressing a single button. • The microscope recognizes the objective chosen and the respective contrast process. Therefore, the values for intensity (INT), aperture diaphragm (AP) and field diaphragm (FD) are always set correctly. • The values for INT, AP and FD can be changed individually.
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6. Startup • Switch-on the microscope at the power switch (34.1,36.1). All motorized microscope components first undergo an initialization phase. After initialization is complete, the display on the stand shows the current microscope setting (Fig. 37). After turning on the gas discharge lamps, 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. 6.
6. Startup The display shows the current microscope settings. The display depends on the microscope’s configuration. In the first column, corresponding pictograms indicate the type of information: contrast method, magnification, light intensity, diaphragms, light splitting for photo tubes. Please see the abbreviation index for a list of abbreviations and pictograms used → p. 67. Light Intensity The actual brightness setting is graphically depicted by a beam.
6. Startup There is a row of function keys both on the right and left side of the stand. Some of these keys are defined, and some of them are variable. The variable function keys have various meanings depending on the microscope configuration. Defined Function Keys on the left side of the stand The TL/IL key (38.1) switches between incident light and transmitted light. The last contrast method used is restored. The INT (38.3) keys adjust the light intensity individually.
6. Startup However, depending on how the condenser is disassembled and reassembled, it may be necessary to re-adjust the condenser in some cases. Therefore, check the condenser centering. The following procedure is provided for the transmitted light-bright field illumination. • Select an objective magnification (10x-40x). with moderate • Activate the transmitted light axis by pushing the TL/IL button (38.1). "TL" appears in the first line of the display.
6. Startup • Open the field diaphragm just enough for it to disappear from the field of view (41d). Do not adjust the aperture diaphragm. The aperture diaphragm is already set optimally for each objective. 6.6. Checking Phase Contrast Rings Caution: Fig. 40 Condenser centering 1 Centering bolts 1 38 Fig.
6. Startup If your microscope is equipped for the use of phase contrast, the light rings that fit the objectives are built into the condenser. The light rings are already leveled in the factory. However, the leveling should be rechecked. • In the place of an eyepiece, insert the focusing telescope (Fig. 42) into the observation tube. • Swivel in the phase contrast objective with the least magnification. Note: Every objective is assigned its own light ring in the condenser disc.
6. Startup • Insert the centering key through the corresponding openings (44.1) in the condenser holder. • Turn the centering screws until the dark ring (phase ring in the objective) is congruent with the slightly narrower bright ring (light ring in condenser) (43 c). • Repeat the process for all other phase contrast objectives. • Remove the centering keys after the centering procedure. Note: During change of objectives the centering keys must not remain in the openings of the condenser. 6.
6. Startup The 107/2 lamp housing with 12 V 100 W halogen lamp has a defined presetting. The lamp need not to be centered. When switching to the BF or Smith reflectors, there is a danger of being glared! • When a supply unit is used, it is turned on first. For the 106 z lamp housing, the direct filament image (for halogen lamps) or direct arc image (for gas discharge lamps), and its mirror image are focused separately and adjusted to each other.
6. Startup • In the adjustment window, you see the direct filament image and the mirror image, which in most cases are shifted together. Fig. 47 Direct lamp filament image focused, but not centered (in reality, the image is less focused) Fig. 48 Direct lamp filament image in target position (in reality, the image is less focused) Fig. 49 Direct lamp filament image and mirror image in target position (in reality, the image is less focused) • Focus the direct filament image with the collector (46.6).
6. Startup Fig. 50 Direct arc image focused but decentered (in reality, the image is less focused) Fig. 51 Direct arc image in target position (in reality, the image is less focused) Fig. 52 Direct arc image and mirror image in target position (in reality, the image is less focused) • Focus the direct image with the collector (46.6). • Use the adjusting buttons on the rear side of the lamp housing (46.2,46.4) to pivot the arc’s mirror image to the side or completely out of the beam path.
6. Startup Fig. 53 Direct arc image focused but not centered (in reality, the image is less focused) Fig. 54 Direct arc image in target position (in reality, the image is less focused) Fig. 55 Direct arc image and mirror image in target position (in reality, the image is less focused) • Focus the direct image with the collector (46.6). • Use the adjusting buttons to pivot the arc’s mirror image on the rear side of the lamp housing (46.2,46.4) to the side or completely out of the beam path.
6. Startup In older lamps, the structure of the arc is no longer clearly recognizable. The image is then more like that of a HG 50 lamp. The image and mirror image can no longer be superimposed exactly. In this case, align both images. • Using the collector, defocus the image with the knob (46.6) until the arc image and mirror image are no longer recognizable and the image is uniformly illuminated. • Exchange the reflector cube for lamp adjustment for the original filter cube.
7. Operation 7. Operation 7.1 Switching on the Microscope When using a gas discharge lamp, the ebq 100 external supply unit must be turned on separately (56.1). Then switch-on the microscope at the power switch. All motorized microscope components first undergo an initialization phase. After the initialization is complete, the display on the stand (Fig. 57) shows the current microscope setting. 7.2 Stages and Specimen Displacement Lengthening the coaxial pinion • For lengthening, pull the lower grip (58.
7. Operation Rotating the Stage 7.3 Focusing The swiveling range of the rotating stages is 0°- 110°. There is a focus dial on the left side of the stage for coarse and fine focus adjustment (Fig. 59). • In order to revolve the stage, loosen the fastening screw (59.1). On the right side of the stand, there is also a focus dial, which is used exclusively for fine focusing (58.4).
7. Operation 7.4 Tubes Adjusting the Viewing Angle • For the AET22 and EDT22 ergotubes, the viewing angle can be adjusted by tilting the binocular viewer in the range of 5° - 32° (Fig. 61). Note: Close any unused tube openings, as otherwise stray light can interfere with observation. Note: Make sure that the connector cable is plugged in on the MBDT25+ motorized tube (60.1). Adjusting the Eyepiece Extension to the Arm Length • With the AET22 tube, the eyepieces can be extended up to 30 mm (Fig. 61).
7. Operation Beam Splitting in Photo Tubes 7.5 Eyepieces EDT22 tube: The beam splitting between the observation and documentation outputs has a definite presetting (50:50). BDT25+ tube: The beam splitting is set manually by pulling out a control bar. Control Bar VIS 50/50 PHOTO Observation 100 % 150 % 110 % Photo 0% 50 % 100 % MBDT25+ tube: This tube is similar to the documentation tube BDT25+, but it is motorized. The control positions are selected using a variable function key on the stand.
7. Operation 7.6 Objectives The objective must be moved manually into the light path. Be sure that the nosepiece turret locks into place. The objective’s position in the turret is factoryset and must be adhered to while screwing in the objectives (see Objective Assembly → p.
7. Operation For lockable immersion objectives: 7.7 Magnification Changer • Lock these by pushing the front part upwards until it stops (approx. 2 mm). Optionally, a coded magnification changer can be used, which is manually operated. On the knurled ring, the following magnification factors can be set: For objectives with corrective mounts: • Turn the knurl to adjust the objective to the thickness of the cover glass. Immersion objective (released) ↔ Fig. 63 B Stand 1x 1.25x 1.6x M Stand 1x 1.
7. Operation 7.8 Light Sources 7.9 Aperture Diaphragm and Field Diaphragm • The brightness is set using the function keys (65.5). Then, the INT function keys are assigned to the currently active axis for transmitted light (TL) or incident light (IL). Both diaphragms are already factory-set to the optimum setting for the current objective. • For TL and IL: Settings can be made either in large or small increments. Pushing both INT buttons simultaneously switches between coarse and fine setting.
8. Imaging Procedure for Leica DM4000 B/DM5000 B 8. Imaging Procedure for Leica DM4000 B/ Leica DM5000 B 8.1 Transmitted Light 8.1.1 Bright Field (TL) • Switch to the transmitted light axis (TL) by pushing the TL/IL button. • Select the BF (bright field) contrast method. Do so by pressing the BF variable key. Alternatively: Press the CHANGE TL | variable key. (For key occupation please see “Identification Sheet”.) The display indicates BF. • Insert a transmitted light specimen.
8. Imaging Procedure for Leica DM4000 B/DM5000 B 8.1.3 Dark Field (TL) Notes: • The microscope automatically selects the correct light ring in the condenser. • When selecting the phase contrast method, the aperture diaphragm is opened completely and may not be adjusted. To avoid errors in operation, the function keys for setting the aperture diaphragm (AP) are locked. • Switch to the transmitted light axis (TL) by pushing the TL/IL button. • Select the DF (dark field) contrast method.
8. Imaging Procedure for Leica DM4000 B/DM5000 B 8.1.4 Polarization (TL) • Switch to the transmitted light axis (TL) by pushing the TL/IL button. • Select the POL (polarization) contrast method. Do so by pressing the POL variable key. Alternatively: Press the CHANGE TL | variable key. (For key occupation please see “Identification Sheet”.) The display indicates POL. Mechanical procedure: • Turn the polarizer on the underside of the condenser in the light path (Fig. 66).
8. Imaging Procedure for Leica DM4000 B/DM5000 B 8.1.5 Differential Interference Contrast (TL) (only for DM5000 B) • Switch to the transmitted light axis (TL) by pushing the TL/IL button. • Insert a specimen and rotate a suitable objective into place. • Select the DIC contrast method. Do so by pressing the DIC variable key. Alternatively: Press the CHANGE TL | variable key. (For key occupation please see “Identification Sheet”.) The display indicates ICT.
8. Imaging Procedure for Leica DM4000 B/DM5000 B 8.2 Fluorescence • Switch to the fluorescent light axis (FLUO) by pushing the TL/IL button. • Insert a specimen and rotate a suitable objective into place. • The current fluorescence cube is indicated on the display. • Closing the incident light shutter protects your specimen from fading. Do so by pressing the SHUTTER variable key. (For key occupation please see “Identification Sheet”.
9. Imaging Procedure for Leica DM4000 M 9. Imaging procedure for Leica DM4000 M 9.1 Incident Light 9.1.2 Dark Field 9.1.1 Bright Field • Switch to the incident light axis (IL) by pushing the TL/IL button. • Switch to the incident light axis (IL) by pushing the TL/IL button. • Select the BF (bright field) contrast method. Do so by pressing the BF variable key. Alternatively: Press the CHANGE RL | variable key. (For key occupation please see “Identification Sheet”.) The display indicates BF.
9. Imaging Procedure for Leica DM4000 M 9.1.3 Polarization Automatic procedure: • Switch to the incident light axis (IL) by pushing the TL/IL button. • Select the POL (polarization) contrast method. Do so by pressing the POL variable key. Alternatively: Press the CHANGE RL | variable key. (For key occupation please see “Identification Sheet”.) The display indicates POL. • The ICR filter cube is automatically brought into the light path. Mechanical procedure: • Rotate the appropriate polarizer (71.
9. Imaging Procedure for Leica DM4000 M 9.1.4 Interference Contrast 9.2 Transmitted Light • Switch to the incident light axis (IL) by pushing the TL/IL button. 9.2.1 Bright Field • Insert a specimen and rotate a suitable objective into place. • Select the DIC contrast method. Do so by pressing the DIC variable key. Alternatively: Press the CHANGE RL | variable key. (For key occupation please see “Identification Sheet”.) The display indicates ICR.
10. Trouble Shooting 10. Trouble Shooting Problem Cause/Remedy Stand The microscope does not respond. Make sure that voltage is impressed. Make sure that the microscope is connected to the power supply. Check the cable connections. Inform service technician to change the fuses. Illumination The image is completely dark. Open the shutter (→ p. 35). Check the connection of the lamp houses to the microscope.
10. Trouble Shooting Problem Cause/Remedy Bright Field The specimen can not be brought into focus. Use the correct immersion medium. Lay the specimen with the cover glass towards the top. Make sure that the cover glass thickness is correct and that is conform to the indication on the objective. Check the condenser centering. Dark Field No definite DF contrast is possible. Be sure that a DF objective is being used. The objective aperture setting is too high (maximum 0.75).
10. Trouble Shooting Problem Cause/Remedy Polarization No polarization contrast is possible. Bring the polarizer and analyzer into cross position until they reach maximum darkness (without specimen) (→ p. 55, 59). Fluorescence The image is completely dark (no fluorescence). Open the shutter (→ p. 57). Select the incident light axis (IL) (→ p. 36). Check the antigen-antibody combination. The fluorescence is too weak. Insert the Booster Lens (→ p. 29). Center the lamp (→ p. 41ff).
11. Care of the Microscope 11. Care of the Microscope Caution! Unplug the power supply before performing cleaning and maintenance work! Protect electrical components from moisture! ! Caution: Residual fiber and dust can create unwanted background fluorescence. Cleaning Coated Parts Microscopes in warm and warm-damp climatic zones require special care in order to prevent fungus contamination. The microscope should be cleaned after each use, and the microscope optics should be kept strictly clean. 11.
11. Care of the Microscope Cleaning Glass Surfaces Removing Immersion Oil Remove dust on glass surfaces with a fine, dry and fat-free hair brush, by blowing with a blow bag or vacuum suction. Caution! Follow safety instructions for immersion oil! Carefully remove stubborn dirt on glass surfaces with a clean cloth moistened with distilled water. If the dirt still can not be removed, use pure alcohol, chloroform or benzine.
12. Wear and Spare Parts 12. Essential Wear and Spare Parts Order No. Material No. Replacement Lamp 500 974 500 137 500 138 500 321 500 139 Name Used for Halogen lamp 12 V 100 W High-pressure mercury burner 50 W High-pressure mercury burner 100 W High-pressure mercury burner 100 W (103 W/2) High-pressure xenon burner 75 W 107/2 lamp housing 106 z lamp housing 106 z lamp housing 106 z lamp housing Screw cap for unused objective receptacles 020-422.
13. Abbreviations and Pictograms 13.
14. Index 14.
15. EU Declaration of Conformity 15.
