Leica DM4000M manual

1 Leica DM4000 B Leica DM4000 M Leica DM5000 B Operating Manual[...]

2 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 Phone +49(0)6441-292261 Fax +49(0)6441-292255 In case of questions, please contact: E-mail: MQM-Hotline@leica-microsystems.com[...]

3 Leica DM4000 B Leica DM4000 M Leica DM5000 B Operating Manual[...]

4 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 (includ- ing electronic systems) is not allowed without express written permission from Leica Microsystems Wetzlar GmbH. The term "Wind[...]

5 Contents 6. Startup ........................................................ 31 6.1 Functional Principle .................................. 31 6.2 Switching on the Microscope ................ 34 6.3 The Display (Leica DM4000 B/DM4000 M) ................. 35 6.4 The Function Keys .................................... 36 6.5 Köhler Illumination ...[...]

6 Contents 8.2 Fluorescence ............................................. 57 9. Imaging Procedure for Leica DM4000 M ....................................... 58 9.1 Incident Light ............................................. 58 9.1.1 Bright Field ...................................... 58 9.1.2 Dark Field ......................................... 58[...]

7 1. Important Notes about this Manual (1.2) → p. 20 ! * Numbers in parentheses, such as "(1.2)", corre- spond to illustrations (in the example, Figure 1, Item 2). Numbers with pointer arrows (for example → p.20), point to a certain page of this manual. Special safety instructions are indicated with the triangle symbol shown here, and[...]

8 2. Safety Notes 2. Safety Notes 2.2 Electrical Safety General specifications 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: Microscope For indoor use only. Supply voltage: Frequency: Power input: DM4000[...]

9 2. Safety Notes Caution! Never use any fuses as replacements other than those of the types and the current rat- ings listed here. Using patched fuses or bridging the fuse holder is not permitted. Caution! The microscope’s electrical accessory com- ponents are not protected against water. Water can cause electric shock. Caution! Protect the micr[...]

10 3. Overview of the Instrument Specification Imaging Procedure Transmitted Light Axis Incident Light Axis Z Pinion Objective nosepiece X/Y Stage Tube 3. Overview of the Instrument Leica DM4000 B / DM5000 B • transmitted light: BF, DF, PH, Pol (DM5000 B also ICT) • incident light: fluorescence • integrated into the stand • motorized 5x fil[...]

11 3. Overview of the Instrument Specification Condenser Magnification Changer Control Panels Computer Interface Software Tools CTR5000 Electronics Box Leica DM4000 B / DM5000 B • manual • absolute coded • 1x; 1.25x; 1.6x For Leica DM5000 B only: Separate control unit with power supply for 100W halogen lamp see p. 8 (electrical safety) Leica [...]

12 3. Overview of the Instrument 1 Eyepiece 2 Eyepiece tube 3 Tube 4 Objective nosepiece with objectives 5 Specimen stage with specimen holder 6 Condenser 7 LC display 8 Function keys field diaphragm 9 Transmitted light/incident light switch 10 Function keys aperture diaphragm 11 Function keys: Light intensity 12 Focus dial with coarse and fine adj[...]

13 3. Overview of the Instrument 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 (factory pre-assigned) 20 X/Y coaxial drive, height adjustable 21 Focus fine adjustment 22 Motorized filter cube exchanger Fig. 2 Leica DM4000[...]

14 4. Unpacking the Microscope The device is delivered in two boxes. The stand box contains the following compo- nents: • Stand with integrated incident light axis and objective nosepiece • Specimen stage with stage bracket • Power cable and PC connecting cable • CD with Leica software package • Instructions and list of microscope default[...]

15 4. Unpacking the Microscope Installation location 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 m[...]

16 5. Assembly 5. Assembling the Microscope T he microscope components are logically as- sembled 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 in- cluded in the delivery package. When usi[...]

17 5. Assembly • From above, set the stage clamp onto the dovetail guide (4.2) and push the stage down- wards 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 leve[...]

18 5. Assembly 5.2 Condenser • Using the condenser height adjuster (5.4), turn the condenser holder (5.1) completely down- wards. • Unscrew the clamping screw for the con- denser (5.3) far enough so that the condenser can be inserted from the front. • From the front, insert the condenser into the condenser holder as far as it will go. On the [...]

19 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). • 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[...]

20 5. Assembly 5.5 Light Sources for the Transmitted Light Axis Caution: Be sure that the lamp housing is discon- nected from the power supply. Unplug the power plug and the power supply during as- sembly. 107/2 Lamp Housing This lamp housing is used with a 12V 100W halo- gen lamp, which is already mounted. In case the lamp has to be removed: • R[...]

21 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. Never look directly into the beam path (blind- ing hazard). During assembly work on xenon burners, al- ways wear the supplied[...]

22 5. Assembly Fig. 17 Lamp mount with 12 V 100 W halogen lamp • Insert the lamp with the dust cover straight into the socket until it stops. • Remove the dust cover. • Reinsert the lamp mount and retighten the fas- tening screw (16.8). Caution: Do not remove the lamp’s dust cover until af- ter you have installed the lamp. Be certain to avo[...]

23 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. 19): Type Typical bulb life* 50 W high-pre[...]

24 5. Assembly Caution: Hg 50 burner: After installation, the labeling must be upright . If a glass melt nipple is present (19a.4), posi- tion it by turning the burner so that the nipple does not come in the way of the beam path later, but instead is positioned sideways. Xe 75 burner: Remove the burner’s dust cover (19b.5) after you have installe[...]

25 5. Assembly Fig. 22 Rear side of the ebq 100 supply unit 1 Lamp connection • Insert the lamp mount, with the burner in- stalled, 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 hous[...]

26 5. Assembly Fig. 25 Removing the front panel 1 Filter receptacle 2 Retention pin 3 Front panel Fig. 26 Inserting the filter or reflector cubes 1 Mounting 1 2 Fig. 23 Filter cube front side 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[...]

27 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). • 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 polar[...]

28 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 sec- tion 5.7 "Equipping the Incident Light filter turret" → p. 26, in the corresponding position on the filter turret. Se[...]

29 5. Assembly Ergomodule For raising the eye level of the tube opening, the ergomodule may be used. It is fastened in place with the side clamping screw. 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. • Place the lamp housing onto the mir[...]

30 5. Assembly Fig. 34 Rear side of stand Leica DM4000 B/M 1 Power switch 2 Power supply 1 2 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 Only for the Leica DM5000 B: • Connect the microscope (36.1) to the "Microscope" jack (35[...]

31 6. Startup 6. Startup 6.1 Functional Principle The microsc ope’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 micro scope recognizes the objective chosen and the respective contrast process. There- fore, the values[...]

32 6. Startup Possible Assignments for the Function Keys For Leica DM4000 B/DM5000 B: Function key Meaning BF Bright field (Transmitted light) PH Phase contrast (Transmitted light) ICT Interference contrast (Transmitted light) DF Dark field (Transmitted light) POL Polarization (Transmitted light) CHANGE TL Switch through all transmitted light proce[...]

33 6. Startup For Leica DM4000 M: Function key Meaning BF Bright field (Incident light) ICR Interference contrast (Incident light) DF Dark field (Incident light) POL Polarization (Incident light) CHANGE RL Switch through all incident light processes INT ↑ Increase brightness ( incident light) INT ↓ Reduce brightness (incident light) AP ↑ Open[...]

34 6. Startup Fig. 37 Display after initialization • 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). The microscopic components such as dia- phragms, conden[...]

35 6. Startup The display shows the current microscope set- tings. The display depends on the microscope’s configuration. In the first column, corresponding pictograms indicate the type of information: con- trast method, magnification, light intensity, dia- phragms, light splitting for photo tubes. Please see the abbreviation index for a list of [...]

36 6. Startup Variable function keys: A factory preset is performed which fits your mi croscope configuration. The function keys are labeled accordingly, and a separate description of the key occupation accompanies the microscope (“Identification Sheet”). Abbreations are listed on p.32f. 6.5 Köhler Illumination For each objective, optimal valu[...]

37 6. Startup However, depending on how the condenser is disassembled and reassembled, it may be nec- essary 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 with moderate magnification (10x-40x). • A[...]

38 6. Startup • Open the field diaphragm just enough for it to disappear from the field of view (41d). Caution: Do not adjust the aperture diaphragm. The aper- ture diaphragm is already set optimally for each objective. 6.6. Checking Phase Contrast Rings A B CD Fig. 41 Köhler Illumination a Field diaphragm not focused, not centered b Field diaph[...]

39 6. Startup If your microscope is equipped for the use of phase contrast, the light rings that fit the objec- tives are built into the condenser. The light rings are already leveled in the factory. However, the leveling should be rechecked. Note: Every objective is assigned its own light ring in the condenser disc. Therefore, a check must be perf[...]

40 6. Startup • Insert the centering key through the corre- sponding 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 con- trast objectives. • Re[...]

41 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. Incident light axis (IL) with 106 z lamp housing • When a supply unit is used, it is turned on first. • Activate the incident light axis using the TL/IL function key. FLUO (Leica DM4000 B/ DM5000 B) or IL (Leica DM4000 M[...]

42 6. Startup • In the adjustment window, you see the direct filament image and the mirror image, which in most cases are shifted together. • Focus the direct filament image with the col- lector (46.6). • Use the adjusting buttons on the rear side of the lamp housing (46.2, 46.4) to pivot the lamp filament’s mirror image to the side or com-[...]

43 6. Startup • 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. The lamp filament’s focused image remains visible (Fig. 50). • Use the adjusting buttons (46.1) and (46.5) to place t[...]

44 6. Startup 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. The arc’s focused [...]

45 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 im- age and mirror image can no longer be su- perimposed 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[...]

46 7. Operation Fig. 58 Revolving object stage 1 Object motion (Y direction) 2 Object motion (X direction) 3 Torque adjustment (Y direction) 4 Torque adjustment (X direction) 5 Focus dial for fine focusing 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.[...]

47 7. Operation Rotating the Stage The swiveling range of the rotating stages is 0°- 110°. • In order to revolve the stage, loosen the fas- tening screw (59.1). • Bring the table into the desired position. • Retighten the fastening screw. Fig. 59 Revolving object stage 1 Clamping screw 2 Fine focusing 3 Coarse focusing 1 7.3 Focusing There [...]

48 7. Operation 7.4 Tubes 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 Viewing Distance • Adjust the viewing distance of the eye- pieces so that a congruent total image is seen (Fig. 60). Fi[...]

49 7. Operation Beam Splitting in Photo Tubes 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 Observation Photo VI S 100 % 0 % 50/50 1 50 % 50 % PHOTO 11 0 % 100 % MBDT25+ tube: This tube is s[...]

50 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 factory- set and must be adhered to while screwing in the objectives (see Objective Assembly → p. 19) When you rotate the objective into position, the microscope a[...]

51 7. Operation 7.7 Magnification Changer Optionally, a coded magnification changer can be used, which is manually operated. On the knurled ring, the following magnification factors can be set: B Stand M Stand 1x 1x 1.25x 1.5x 1.6x 2x The selected factor is indicated in the display and included in the total magnification. Fig. 63 Immersion objectiv[...]

52 7. Operation 7.8 Light Sources • The brightness is set using the function keys (65.5). Then, the INT function keys are as- signed to the currently active axis for trans- mitted light (TL) or incident light (IL). • For TL and IL: Settings can be made either in large or small increments. Pushing both INT buttons simultaneously switches between[...]

53 8. Imaging Procedure for Leica DM4000 B/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 “Ident[...]

54 8. Imaging Procedure for Leica DM4000 B/DM5000 B 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 ap- erture diaphragm (AP) are [...]

55 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”.)[...]

56 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 objec- tive into place. • Select the DIC contrast method. Do so by pressing the DIC variable key. Alternatively[...]

57 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 objec- tive into place. • The current fluorescence cube is indicated on the display. • Closing the incident light shutter protects your specimen from fading[...]

58 9. Imaging Procedure for Leica DM4000 M 9.1 Incident Light 9.1.1 Bright Field • 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”.)[...]

59 9. Imaging Procedure for Leica DM4000 M 9.1.3 Polarization • 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 indi[...]

60 9. Imaging Procedure for Leica DM4000 M 9.1.4 Interference Contrast • Switch to the incident light axis (IL) by pushing the TL/IL button. • Insert a specimen and rotate a suitable objec- tive into place. • Select the DIC contrast method. Do so by pressing the DIC variable key. Alternatively: Press the CHANGE RL variable key. (For key occup[...]

61 10. Trouble Shooting 10. Trouble Shooting Problem Stand The microscope does not respond. Illumination The image is completely dark. The image is unevenly or not uniformly illumi- nated. The illumination "flickers." The lamp does not illuminate immediately upon being switched on. Cause/Remedy  Make sure that voltage is impressed. [...]

62 10. Trouble Shooting Problem Bright Field The specimen can not be brought into focus. Dark Field No definite DF contrast is possible. The image is unevenly or not uniformly illumi- nated. Undesirable stray light Phase contrast No phase contrast is possible. Cause/Remedy  Use the correct immersion medium.  Lay the specimen with the cover gl[...]

63 10. Trouble Shooting Problem Polarization No polarization contrast is possible. Fluorescence The image is completely dark (no fluorescence). The fluorescence is too weak. Display The display flashes. FAIL! appears. Cause/Remedy  Bring the polarizer and analyzer into cross po- sition until they reach maximum darkness (without specimen) ( → p[...]

64 11. Care of the Microscope Caution! Unplug the power supply before performing cleaning and maintenance work! Protect electrical components from moisture! 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[...]

65 11. Care of the Microscope Cleaning Glass Surfaces Remove dust on glass surfaces with a fine, dry and fat-free hair brush, by blowing with a blow bag or vacuum suction. 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.[...]

66 12. Wear and Spare Parts Order No. Material No. Name Used for Replacement Lamp 500 974 Halogen lamp 12 V 100 W 107/2 lamp housing 500 137 High-pressure mercury burner 50 W 106 z lamp housing 500 138 High-pressure mercury burner 100 W 106 z lamp housing 500 321 High-pressure mercury burner 100 W 106 z lamp housing (103 W/2) 500 139 High-pressure [...]

67 13. Abbreviations and Pictograms 13. Abbreviations and Pictograms Contrast method Magnification Light intensity/diaphragms Beam splitting Transmitted light shutter open Transmitted light shutter closed Incident light shutter open Incident light shutter closed ↓ ↓ ↑ ↑ +   AET Advanced Ergo Tube AP Aperture diaphragm BF Brifght field[...]

68 14. Index A djusting the light sources 41 Allowable ambient conditions 15 Ambient temperature 8, 9 Ambient conditions 15 Analyzer 28, 55, 56, 59 Analyzer cube 55, 56 Aperture diaphragm 12, 35, 38, 52 B eam splitting 49 Bright field 53, 58 Booster Lens 29, 5 7 C leaning 64 Cleaning objectives 65 Coaxial pinion 46 Condenser connector 18 Condenser [...]

69 15. EU Declaration of Conformity 15. EU Declaration of Conformity[...]