Leica DM4000B, Operating Manual

Page 1: ...1 Leica DM4000B Leica DM4000M Leica DM5000B Operating Manual ...

Page 2: ...Leitz Straße D 35578 Wetzlar Germany Responsible for contents Katja Peter Karin Schwab Marketing CM Compound Microscopy ProductManagement Phone 49 0 6441 292261 Fax 49 0 6441 292255 In case of questions please contact E mail MQM Hotline leica microsystems com ...

Page 3: ...3 Leica DM4000B Leica DM4000M Leica DM5000B Operating Manual ...

Page 4: ...the Microsoft Corporation Otherwise no inference with regard to the free usability of product names may be drawn from the use of those names The instructions contained in the following docu mentation reflect state of the art techno logy 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 accurac...

Page 5: ... 53 8 1 Transmitted Light 53 8 1 1 Bright Field 53 8 1 2 Phase Contrast 53 8 1 3 Dark Field 54 8 1 4 Polarization 55 8 1 5 Differential Interference Contrast 56 Contents 1 Important Notes about this Manual 7 2 Safety Notes 8 2 1 General Safety Notes 8 2 2 Electrical Safety 8 3 Overview of the Instrument 10 4 Unpacking the Microscope 14 5 Assembling the Microscope 16 5 1 Stage 17 5 2 Condenser 18 5...

Page 6: ...9 1 3 Polarization 59 9 1 4 Interference Contrast 60 9 2 Transmitted Light 60 9 2 1 Bright Field 60 10 Trouble Shooting 61 11 Care of the Microscope 64 11 1 Dust Cover 64 11 2 Cleaning 64 11 3 Handling Acids and Bases 65 12 Essential Wear and Spare Parts 66 13 Abbreviations and Pictograms 67 14 Index 68 15 EU Declaration of Conformity 69 ...

Page 7: ...he microscope and accessories can be damaged when operated incorrectly Explanatory note Item not contained in all configurations Text symbols and their meanings Caution This operating manual is an essential com ponent of the microscope and must be read carefully before the microscope is put into operation or used 1 Important Notes about this Manual This operating manual contains important in struc...

Page 8: ...trol and laboratory devices Caution In order to maintain this condition and to en sure 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 must be consulted when ever th...

Page 9: ...y unit ebq 100 For indoor use only Supply voltage Frequency Power input Fuses Ambient temperature Relative humidity Overvoltage category Pollution degree see enclosed manual 90 250 V 50 60 Hz max 155 VA 2xT2A IEC 127 15 35 C max 80 to 30 C II 2 Caution 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 extens...

Page 10: ...lute coded 6x with M25 thread DM5000 B 7x mot DIC objective prism turret with 4 positions optional manual replaceable specimen stage coaxial pinion length 155 mm Leica DM4000 M transmitted BF DF PH light ICT Pol incident light BF DF ICR Pol integrated into the stand motorized 4x filter turret automatic Illumination Manager motorized shutter manual absolute encoded 6x with M32 thread slot for DIC p...

Page 11: ...lectrical safety Leica DM4000 M manual absolute coded 1x 1 5x 2x motorized condenser head motorized condenser turret for light rings DF stop DIC prisms optional polarizer integrated and motorized automatic Köhler Illumination operating buttons for all motorized microscope functions additional variable function keys focusing knobs LC display DM5000 B with LeicaScreen touchscreen RS232C Leica DMCont...

Page 12: ... 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 adjustment 13 Variable function keys factory pre assigned 14 Lamp adjustment window Fig 1 Leica DM4000 M left side of the stand with AET22 advanced ergotube 1 2 3 4 5 6 7 8 9 10 11 12 13 14 ...

Page 13: ...ansmitted 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 B right side of the stand with Advanced Ergotube AET22 22 21 20 19 18 17 15 16 ...

Page 14: ...e accessories such as filter cubes etc 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 Note Avoid touching the lens surfaces of the objectives If fingerprints do appear on the glass surfaces r...

Page 15: ...rm damp climatic zones require special care in order to prevent the build up of fungus See the chapter Care of the microscope p 64 for additional instructions Caution Electrical components must be assembled at least 10 cm from the wall and away from flammable substances Transport For shipping or transporting the microscope and its accessory components the original packaging should be used As a pre...

Page 16: ...ser 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 using intermediate systems and optical accessories the sequence may vary In this case read Chapter 5 10 Optional accessories p 29 ...

Page 17: ...ingly lower level 1 2 Fig 4 Assembling the stage 1 Stage clamp 2 Dovetail guide 5 1 Stage Caution Before assembling the stage make sure no ob jectives are installed 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...

Page 18: ...n tation pin 6 1 which must be located in the guiding notch 7 1 Pull the condenser s clamping screw 5 3 so that the condenser is locked in place Connect the condenser over the connection 8 1 with the stand Fig 5 Condenser holder 1 Condenser holder 2 Condenser centering 3 Clamping screw for condenser 4 Condenser height adjuster Fig 6 Underside of condenser 1 Orientation pin Fig 7 Condenser holder 1...

Page 19: ...nector socket 10 1 The eyepieces are inserted into the eyepiece tubes on the tube 6 4 Objectives The receptacles on the objective turrets are numbered Fig 11 The individual objectives have already pre assigned positions at the factory according to their configuration A list of the exact objective positions is provided in shipment Identification Sheet Caution Cover unoccupied threads on the turret ...

Page 20: ...ight Remove the lamp s dust cover Caution Do not remove the lamp s dust cover until you have installed the lamp Avoid fingerprints on the lamp Replace the housing and fasten it in place us ing the fastening screw Place the lamp housing in the transmitted light lamp housing receptacle 14 2 and fasten it with the clamping screw on the side Connect the lamp housing to the power supplyfor transmitted ...

Page 21: ...z lamp housing This lamp housing is used with a 12V 100W halo gen 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 8 and pull out the mount Fig 17 Fig 15 Protective gloves and mask Fig 16 106 z lamp housing on the side open 1 Cove...

Page 22: ...o avoid getting fingerprints on the lamp Close the lamp housing and retighten the fas tening screws Place the lamp housing in the incident light lamp housing receptacle 18 1 and fasten it with the clamping screw on the side Connect the lamp housing to the power supply for incident light symbol 18 4 1 2 3 4 Fig 18 Rear side of stand 1 Incident light lamp housing receptacle 2 Transmitted light lamp ...

Page 23: ...harge lamps may be used and require different lamp mounts Fig 19 Type Typical bulb life 50 W high pressure mercury burner alternating current 100 hrs 100 W high pressure mercury burner direct current stabilized not stabilized 200 hrs 100 W high pressure mercury burner direct current stabilized not stabilized type 103 W 2 300 hrs 75 W High pressure xenon burner direct current stabilized 400 hrs Ple...

Page 24: ...ew the fastening screws on the cover 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 Install the burner in mirror image fashion Fig 19 a d Lamp mounts for gas discharge lamps 1 Upper clamping syst...

Page 25: ...ith the clamping screw on the side Connect the lamp housing to the power supply 22 1 1 Fig 20 106 z lamp housing on the side open 1 Cover raised 2 Collector 3 12 V 100 W lamp or gas discharge lamp in mount 4 Reflector mirror 5 6 7 Adjusting screw for x y reflector 8 Fastening screw for lamp mount 9 Socket for contact plug 1 2 3 4 1 2 3 8 9 8 4 5 6 7 Fig 21 Rear side of stand 1 Incident light lamp ...

Page 26: ...ope is switched off Remove the face plate from the upper part of the microscope Fig 25 Turn the turret in any direction until the locking pin engages Insert the filter or reflector cube into the mounting in front of you according to the identification sheet provided To do so place the filter or reflector cube on the right side and press it to the left into the mounting Fig 26 Push the retention pi...

Page 27: ...t light polarizers R P polarizer rotating polarizer L ICR R ICR polarizer Remove the plug cap on the right side of the incident light axis Fig 29 Insert the polarizer into the receptacle until it latches in place Motorized polarizer A motorized polarizer is already installed and ready for operation in the DIC condenser Transmitted light and incident light analyzer Fig 27 Assembly of the ICT P tran...

Page 28: ...ee 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 Insert the objective prism into the tube slot Fig 31 1 The code letter must match the code letter on the objective With the microscope Leica DM5000 B the DIC objective prisms are already mounted in the DIC turret above the objective revolving ...

Page 29: ...rror hous ing and fasten it using the corresponding clamping screw on the side Booster Lens Excitation Manager Fig 32 1 Insert of Booster Lens Fig 33 1 Insert of Excitation Manager Remove the clamping ring from the filter slide Insert the Booster Lens or Excitation Manager Push the cover back Insert the clamping ring Insert the filter slide into the front receptacle on the right side of the stand ...

Page 30: ...00 Electronics Box 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 sup ply using the power cable 35 2 Fig 35 Rear side of electronics box CTR5000 1 Microscope connection 2 Power supply Fig 36 Rear side of stand Leica DM5000 B 1 Connection to the CTR5000...

Page 31: ... light axis trans mitted light or incident light In addition to the preset function keys for INT AP and FD there are also variable function keys Variable function keys These function keys are assigned logical functions before delivery see Identification Sheet These functions can be reprogrammed according to your individual wishes Note Reset Function The microscope can be reset to the default funct...

Page 32: ...agm transmitted light FD Open field diaphragm transmitted light FD Close field diaphragm transmitted light SHUTTER TL Open close transmitted light shutter FLUO Fluorescence last filter cube CUBE 1 Select fluorescence cube at position 1 CHANGE CUBE Switch through fluorescence cubes in clockwise fashion CHANGE CUBE Switch through fluorescence cubes in counterclockwise fashion SHUTTER FLUO Open close...

Page 33: ...field diaphragm incident light SHUTTER RL Open close incident light shutter FLUO Fluorescence last filter cube CUBE 1 Select fluorescence cube at position 1 CHANGE FLUO Switch through fluorescence cubes FOCUS FINDER Select smallest field diaphragm and switch back to original field diaphragm by pressing the key again BF TL Bright field Transmitted light INT Increase brightness transmitted light INT...

Page 34: ...denser light and phase rings are already pre centered in the factory However re centering may be necessary due to transpor tation and assembly Before proceeding with the necessary steps first familiarize yourself with the stand s display and control panel Caution After turning on the gas discharge lamps the burner must be immediately adjusted There fore do not turn on the power supply unit yet Fir...

Page 35: ...tion changer appears along with the total magnification Σ Objective x Re magnification x Eyepiece 6 4 The Function Keys Light Intensity The actual brightness setting is graphically de picted by a beam Additionally the light intensity is indicated in 20 coarse adjustment or in 255 fine adjustment increments p 52 Diaphragms The values for the field diaphragm FD and the aperture diaphragm AP are indi...

Page 36: ... 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 indi vidually Settings can be made either in large or small increments Pushing both INT buttons at the same t...

Page 37: ... specimen holder 39 3 Focus on the specimen The focus wheel on the left side of the stage allows focus adjust ment in large and small increments On the right side of the stage there is also a focus wheel for fine focus adjustment Set the light intensity using the INT keys 38 3 Close the field diaphragm with the FD function key 38 2 until the edge of the diaphragm ap pears in the specimen plane Usi...

Page 38: ...eady set optimally for each objective 6 6 Checking Phase Contrast Rings A B C D Fig 41 Köhler Illumination a Field diaphragm not focused not centered b Field diaphragm focused but not centered c Field diaphragm focused and centered Diameter is too small however d Field diameter light Field diameter view Köhler Illumination Fig 40 Condenser centering 1 Centering bolts 1 1 ...

Page 39: ...ast centering procedure PH phase contrast ring LR light ring a Condenser in bright field BF position b Condenser in phase contrast PH position Light ring LR not centered c Light ring and phase ring centered In the place of an eyepiece insert the focus ing telescope Fig 42 into the observation tube Swivel in the phase contrast objective with the least magnification Focus the ring structure 43a by s...

Page 40: ...ightly narrower bright ring light ring in condenser 43 c Repeat the process for all other phase con trast 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 7 Adjusting the Light Sources Transmitted Light Axis TL with 107 2 Lamp housing Fig 44 Light ring centering 1 Clamping screw...

Page 41: ...n in the upper right of the display Caution Never look directly into the light path When switching to the BF or Smith reflectors there is a danger of being glared For the 106z lamp housing the direct filament im age for halogen lamps or direct arc image for gas discharge lamps and its mirror image are focused separately and adjusted to each other On the left side of the microscope there is an adju...

Page 42: ...lign the mirror image symmetrically to the fil ament image Fig 49 To do so use the adjust ing knobs 46 2 and 46 4 again Defocus the image with the collector head 46 6 until the filament image and mirror im age are no longer recognizable and the image is uniformly illuminated Exchange the reflector cube for lamp adjustment for the original filter cube Note Turn off the microscope before exchanging ...

Page 43: ... knobs 46 2 and 4 to orient the mirror image symmetrically to the direct image Fig 52 Defocus the image with the collector 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 Centering the Hg 100 W and Xe 75 W mercury lamps In the adjustment window you see the d...

Page 44: ... the arc the focal spot should lie slightly outside the center Fig 54 Then pivot the arc s mirror image with the ad justing knobs 46 2 and 46 4 and focus it us ing the reflector 46 3 Use the adjusting knobs 46 2 and 4 to orient the mirror image symmetrically to the direct image Fig 55 The V shaped irradiation of the direct image and mirror image arcs can be superimposed Caution The bright tips of ...

Page 45: ...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 longer recognizable and the im age is uniformly illuminated Exchange the reflector cube for lamp adjustment for the original filter cube Note Turn off the microscope before exchanging the reflector cube ...

Page 46: ...motorized microscope components first un dergo an initialization phase After the initialization is complete the display on the stand Fig 57 shows the current microscope setting Fig 57 Display after initialization 7 2 Stages and Specimen Displacement Lengthening the coaxial pinion For lengthening pull the lower grip 58 2 downwards Repeat with the upper grip 58 1 Torque adjustment The torque is alre...

Page 47: ...object stage 1 Clamping screw 2 Fine focusing 3 Coarse focusing 1 7 3 Focusing There is a focus dial on the left side of the stage for coarse and fine focus adjustment Fig 59 On the right side of the stand there is also a focus dial which is used exclusively for fine focusing 58 4 The special design of this dial makes it possible to simultaneously grasp the coaxial drive with your hand while opera...

Page 48: ... of the eye pieces so that a congruent total image is seen Fig 60 Fig 60 Tube setting Personal eyebase settings 1 Motorized tube connection Fig 61 With AET22 tube individual adjustments Adjusting the Viewing Angle For the AET22 and EDT22 ergotubes the view ing angle can be adjusted by tilting the binocu lar viewer in the range of 5 32 Fig 61 Adjusting the Eyepiece Extension to the Arm Length With ...

Page 49: ...iece s aperture protector must be removed or at least folded back during microscopy while wearing eyeglasses Eyeglasses with multifocal lenses bifocals and smooth view glasses must be removed while operating the microscope For the adjustable tubes with documentation output choose the 100 position Eyepieces with Inlaid Reticle Focus the reticle by adjusting the eyelens Focus on the object through t...

Page 50: ...nizes the selected contrast method the optimal settings for field and aperture diaphragm the optimal condenser setting The objective magnification and the total magni fication appear in the display p 35 Start with a small level of magnification Then switch to the next higher objective For immersion objectives use the appropriate immersion medium OIL only use optical immersion oil according to DIN ...

Page 51: ... factor is indicated in the display and included in the total magnification Fig 63 Immersion objective released For lockable immersion objectives Lock these by pushing the front part upwards until it stops approx 2 mm Then after a gentle turning motion to the right the objective is locked Fig 64 For objectives with corrective mounts Turn the knurl to adjust the objective to the thickness of the co...

Page 52: ... 10 Fig 645 Control panel 1 Variable function keys 2 Aperture diaphragm 3 Transmitted light incident light 4 Field diaphragm 5 Light intensity 1 2 3 4 5 7 9 Aperture Diaphragm and Field Diaphragm Both diaphragms are already factory set to the optimum setting for the current objective The AP 65 2 keys for the aperture diaphragm and the FD keys 65 4 for the field diaphragm may be used to change each...

Page 53: ...ocus using the focus dial and set the brightness using the INT function key 8 1 2 Phase Contrast Switch to the transmitted light axis TL by pushing the TL IL button Select the PH contrast phase contrast method Do so by pressing the PH variable key Alternatively Press the CHANGE TL variable key For key occupation please see Identification Sheet The display indicates PH Insert a transmitted light sp...

Page 54: ...cupation please see Identification Sheet The display indicates DF The dark field ring dark field stop is set au tomatically Insert a transmitted light specimen Rotate an appropriate objective into place Bring the image into focus using the focus dial and set the brightness using the INT function key Notes The maximum objective aperture which may be used for dark field is 0 75 All objectives with g...

Page 55: ...ex point on the front of the polarizer is aligned with 0 Insert the analyzer into the left side of the stand 67 1 Bring the polarizer and analyzer into cross po sition until they reach maximum darkness Insert a specimen and rotate a suitable objec tive into place Motorized procedure After selecting the POL contrast method the condenser automatically switches to the posi tion of the polarizer The a...

Page 56: ...ondenser and the fitting condenser prism are automatically brought into the light path The corresponding objective prism and the analyzer cube are also positioned automatically For fine adjustment use the knurled ring above the objective nose piece Fig 68 Fig 68 Objective prism slide 1 Knurled wheel for fine adjusting Alternatively Manually rotate the polarizer on the underside of the condenser in...

Page 57: ...on please see Identification Sheet The display indicates the symbol Selecting the fluorescence filter cube Press the variable keys Cube or Cube The fluorescence intensity can be increased using the Booster Lens on the right side of the stand Fig 69 For multifluorescence use of a Excitation Manager is recommended The Excitation Manager is inserted into the right side of the stand up to the last sto...

Page 58: ... the TL IL button Select the DF dark field contrast method Do so by pressing the DF variable key Alternatively Press the CHANGE RL variable key For key occupation please see Identification Sheet The display indicates DF The DF reflector is turned into the beam path Insert a specimen Rotate an appropriate objective into place Bring the image into focus using the focus dial and set the brightness us...

Page 59: ...L Automatic procedure The ICR filter cube is automatically brought into the light path Mechanical procedure Rotate the appropriate polarizer 71 3 and the IC P analyzer 72 1 on the stand manually into the light path Also bring the polarizer and analyzer into cross position until they reach maximum darkness Insert a specimen and rotate a suitable objec tive into place Fig 72 1 Insert analyzer Fig 71...

Page 60: ...he prism slot 71 2 Alternatively Rotate the ICR polarizer 71 3 and the IC P analyzer 72 1 on the stand manually into the light path Insert the objective prism slide into the prism slot 71 2 For fine adjustment rotate the knurled screw 71 1 on the objective prism slide 9 2 Transmitted Light 9 2 1 Bright Field Switch to the transmitted light axis by pushing the TL IL button Select the BF bright fiel...

Page 61: ... the cable connections Inform service technician to change the fuses Open the shutter p 35 Check the connection of the lamp houses to the microscope Transmitted axis Incident Fluo axis Make sure that the lamps are connected to the power supply Inform service technician to change the fuses of the ebq 100 Remove all unneeded filters from the light path Center the lamp p 41ff Replace the old lamp p 2...

Page 62: ... is correct and that is conform to the indication on the objective Check the condenser centering Be sure that a DF objective is being used The objective aperture setting is too high maximum 0 75 If necessary reduce the ob jective aperture using the iris diaphragm on the objective Check the condenser centering The magnification is too weak Use a higher magnification Clean the specimen and neighbori...

Page 63: ...e Remedy Bring the polarizer and analyzer into cross po sition until they reach maximum darkness without specimen p 55 59 Open the shutter p 57 Select the incident light axis IL p 36 Check the antigen antibody combination Insert the Booster Lens p 29 Center the lamp p 41ff Insert a new lamp p 20f Rotate an appropriate objective for the con trast method into the light path Check insertion of object...

Page 64: ...tion water may occur 11 2 Cleaning Caution Residual fiber and dust can create unwanted background fluorescence Cleaning Coated Parts Dust and loose dirt particles can be removed with a soft brush or lint free cotton cloth Clinging dirt can be cleaned with all commercially available water solutions benzine or alcohol For cleaning coated parts use a linen or leather cloth that is moistened with one ...

Page 65: ...ent to your Leica subsidiary for repair We also advise against cleaning the inside surfaces of the eyepieces The front lenses of objectives are cleaned as de scribed under Cleaning Glass Surfaces The upper lens is cleaned by being blown off with a pneumatic pump Removing Immersion Oil Caution Follow safety instructions for immersion oil First wipe off the immersion oil with a clean cot ton cloth a...

Page 66: ...High pressure xenon burner 75 W 106 z lamp housing Screw cap for unused objective receptacles 020 422 570 000 Screw cap M 25 Objective turret Replacement eyecup diaphragm protection for HC PLAN eyepiece 021 500 017 005 HC PLAN eyecup 10x 25 eyepiece 021 264 520 018 HC PLAN eyecup 10x 22 eyepiece 021 264 520 018 HC PLAN eyecup 10x 20 eyepiece Immersion Oil conforming to DIN ISO standards fluorescen...

Page 67: ...tter closed AET Advanced Ergo Tube AP Aperture diaphragm BF Brifght field COMBI Combination contrast method CUBE Filter cube DF Darkfield DIC Differentiated interference contrast FD Field diaphragm FLUO Fluo axis incident light ICR Interference contrast reflected light ICT Interference contrast transmitted light IL Incident light axis INT Intensity MBDT Motorized Basic Documentation Tube PH Phase ...

Page 68: ...sion oil 50 65 Incident light axis 10 Incident light nosepiece disc 26 Incident light polarizators 27 Incident light shutter 35 Initialization 34 Interference contrast 60 Intermediate systems 16 Köhler illumination 18 37 Lamp housing 13 Lamp housing 106 z 21 41 Lamp housing 107 2 20 41 Lamp housing receptacle 20 22 25 Light intensity 35 Light sources 41 52 Light sources incident light axis 21 Ligh...

Page 69: ...69 15 EU Declaration of Conformity 15 EU Declaration of Conformity ...