Users Manual Fluoview Scanning Laser Biological Microscope Ver. 1.1 WARNING A Caution: Before using your microscope, the items in this manual Identified by the mark shown to the left should be read carefully until completely understood to ensure safe operation. Thank you for purchasing an Olympus Fluoview Microscope. Before using your microscope, read this manual thoroughly to make sure you obtain full performance from all of the functions provided by this system.
raj ?3 n IF=° -About This Manual This manual explains potential risks and provide important safety information designed to ensure the safe use bf the FLUOVIEW System. Thoroughly study this manual before operating the system.
CONTENTS 1 Safety Precautions A 2 Warning Labels 2 - 1 Warning Labels Concerning Laser Safety 2 - 1 - 1 Warning Labels 2 - 1 - 2 Aperture Label 2 - 1 - 3 Protective Housing Label 3 Precautions for Use of the System 1-1 2-1 2-1 2-1 2-3 2-5 3J^
1. Safety Precautions 1 Safety Precautions A Depending on the laser configuration, this laser product is classified as follows: Ar laser configuration — CLASS n OAyilONl LASER RADIATION DO NOT S[fPt IN1D S m ARGON LASER InW Mf\X CW 450-515nn (LASS n LASKPRGDUCT Kr and Ar laser configuration CLASS in a ANGE LASER LIGHT /M)IDD!fB]TBEB«RRE KR. AR LASER SnW i MAX CW 450-570na CLASS l a LASER PRdXJCT Precautions for Use Always pay careful attention to the following points.
1. Safety Precautions If the objective revolving nosepiece or the objective lens is removed during laser output (during laser scanning), the laser beam will be emitted uncontrollably. This is extremely dangerous. Never remove the objective revolving nosepiece or the objective lens. TThe nosepiece or the objective lens is to be removed only by trained personnel, and only when [_ power has been removed from the laser system.
1. Safety Precautions A Please note that the laser beam is emitted when the display's function pane! ([Acquire] panel) looks as shown below. "Laser beam emitted" is shown. w}iFjmt^axv/),^^.\jiu»:p^ Disposal of Laser Tubes Strict adherence to applicable industrial guidelines and laws is required when disposing of laser tubes. Before disposing of laser tubes, contact your Olympus representative for further information.
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2. Warning Labels 2 - 1 - 2 Aperture Label (Indicates where the laser beam is emitted.) (3) AVOID EXPOSURE Laser radiation is emitted from this aperture BX50 Configuration (3) The laser beam is emitted from the objective and the sockets on the revolving nosepiece. (3) The laser beam Is emitted from the condenser lens. BXWI Configuration (3) The laser beam Is emitted from the objective and the sockets on the revolving nosepiece. The laser beam is emitted from the condenser lens.
2. Warning Labels IX Configuration The laser beam is emitted from the objective. (3) (3) 2-4 The laser beam is emitted from the condenser lens.
2. Warning Labels 2 - 1 - 3 Protective Housing Label (5) (4) CAUTION - Laser radiation when open. DO NOT STARE INTO BEAM DANGER - Laser light when open. AVaO DIRECT EYE EXPOSURE. t/-^f3tS®^ajit«L»c t CAUTION - Laser ^ radiation when open. jDO NOT STARE INTO BEAM BX50, BXWI configuration Ar : (4) Kr •» Ar : (5) Do not remove the cube turret. Do not remove the revolving nosepiece, the objective or the DIC slider or the dummy slider.
2. Warning Labels IX configuration Ar : (4) Kr + Ar : (5) Do not remove the cube tuaet. When the transmitted detection unit is not mounted (left) and when mounted (right) Do not remove the transmitted light lamp housing or the lamp socket.
2. Warning Labels Common to all configurations When the slider cover is removed (6) Do not remove the detection mode selector slider. In addition to the locations shown in this manual, a protective housing label is also attached. For replacement of spiled b r p your Olympus representative. 2-7 r warriihg labblSi^ pibaise cohtact ^^^'..' •.:'.' :.'r ••• '••••:;:' •.
3. Precautions for Use of the System 3 Precautions for Use of the System Installation • The FLUOVIEW system will be assembled and setup by qualified technicians. Avoid moving the system as this may adversely affect the adjustment of the optical system. In case it becomes necessary to move the system, please consult your Olympus representative.
3. Precautions for Use of the System • Standard objectives for biological use are employed for both the upright microscope system configuration (BX) and the inverted microscope system configuration (IX). Accordingly, use cover glass with a thickness of 0.17 mm or Petri dishes with a bottom thickness of 0.17 mm. It is recommended to use special cover glass for LSM water immersion objectives.
Concerning This Section This section provides an overview of the Fluoview System and should be read before the system is used. After reading and understanding this section, please read the Operation section.
Contents CAUTIONS 2 REGISTERED TRADEMARKS 2 CARE AND MAINTENANCE 3 MOVING THE FLUOVIEW 3 SECTION 1 INTRODUCTION H 1-1 Fluoview Manual Configuration 1-1 1-1-1 Users Manual Configuration 1-1 1-2 Conventions Used in This Manual SECTION 2 SYSTEM OVERVIEW 1-2 2J.
Cautions (1) This software and manual may not be reproduced in part or in their entirety without the express written permission of Olympus. (2) The contents of this manual are subject to change without notice. Registered Trademarks Microsoft, Microsoft Windows and Excel for Windows are registered trademarks of Microsoft Corporation of the U.S. All other company names and product names are the trademarks or the registered trademarks of the respective companies.
Care and Maintenance (1) When not in use, always use the accessory cover to protect your microscope from dust. (2) This microscope is a precision instrument, so do not disassemble any of the components. (3) Keep dirt, fingerprints off the lenses and filters. Remove any dirt by wiping lightly with soft gauze. Stubborn dirt can be removed by wetting the gauze with a mixture of alcohol and ether (3:7 ratio), or benzene.
Section 1 Introduction Section 1 Introduction 1-1 Fluoview Manual Configuration There are two Fluoview manuals; the Users Manual and the On-screen Manual (on-line help). The Users Manual consists of five sections. The contents of these sections are described below. 1 - 1 - 1 Users Manual Configuration • For Safe Usage This section explains the requests, cautions and wamings related to usage of the Fluoview System. • Introduction to Fluoview This section provides an overview of the Fluoview System.
Section 1 Introduction 1-2 Conventions Used in This Manual The following is an explanation of the various conventions used in this manual. < • Caution, remari< and one-point advisory symbols. Symbol * Explanation Caution items are indicated by (*). Remarks to be observed and one-point advisories are indicated by (@). < • Menu, command button and dialogue box conventions. Usage [Config] panel Explanation The names of panels, dialogue boxes, list boxes, check boxes, etc.
Section 1 Introduction ^ > Terms Unique to This System Convention XY obsen/ation (Other observation) Explanation This means using XY scan to obsen/e. (The same is for XZ, Xt, XVt and XYZt observation.
Section 2 System Overview Section 2 System Overview The Olympus Fluoview is a confocal scanning type laser fluorescent microscope that utilizes a common focal point optical system to realize high resolution and high contrast as well as a spectacular improvement in resolution in the optical axis. This microscope provides researchers with the ability to perform automatic sectioning, three-dimensional structuring and time fluctuation observation as well as various types of image processing and analysis.
Section 2 System Overview 2-2 Fluoview Features 1. The detector has a resolution of 12 bits, so images are extremely clear, 2. 1 A high-resolution 1024 x 768 pixel system is used. Non-interlaced output signals are used for clear, flickeriess images. 3. Transmitted light is detected by a photomultiplier detector, so transmitted light images are sharp and clear. 4. An auto-gain function eliminates any need for bothersome sensitivity adjustment 5.
Section 2 System Overview 2-3 Optical Path Diagram CHI CH2 FL TR Laser CH2 CH2 CHI CH2 TR FL • — CH2 PM2 2-3
Section 2 System Overview 2-4 System Configuration 2-4-1 System Units and Their Roles Scan unit This unit scans the laser beam in the X and Y directions and detects the light that retums. Microscope A BX50 erect image type microscope for fluorescent light Electromagnetic disk observation. A large-capacity storage device for storing images as files. Image monitor Used to display laser scanned images, operation panel, etc.
Section 2 System Overview Microscope A BXWI fixed stage erect image microscope for fluorescent light observation. VIbration-free stand A special pneumatic type stand for eliminating vibration. ^fis^.f •••" Photo 2-2 BXWI System Compressor Supplies air to the vibration-free stand.
Section 2 System Overview Microscope AN 1X70 Inverted Image microscope for fluorescent light observation. Vibration-free stand A special pneumatic type stand for eliminating vibration. Photo 2-3 1X70 System Pt^^ii^ ^W mmM:n^'MF^'^Cz^m^D ij^feg ^r^F P H pjfr-: T* fc^^ RK? ^m . " iri'''**?**SPyt1rtit' ^ - . « " - ^ ' '•"••• t - * . — •^'•nn ' Transmitted light detector (option) This unit Is used to obtain transmitted light images. '|~ " • ' V k ' ^ ' " ^ ' ' * i . .f^^S:% • ^ ^ ^ • :K--<.
Section 2 System Overview 2-5 Software Function Configuration This software uses panel type windows. With conventional software, it is necessary to select a menu and then select the command to be executed. With panels, however, software functions can easily be executed by merely selecting the panel index for the function to be executed, like a system notebook or file folder. 2-5-1 Software Panel Configuration The panel indexes for all of the functions cannot be displayed at one time but must be scrolled.
Section 2 System Overview 2-5-2 Function Panels and Display Panels The Fluoview software utilizes two types of panels: the function panel and the display panel. The functions panel includes the [Acquire], [File I/O], [Tile], [Process], [Analyze], and [Visualize] panels. The [Live] panel and [(File Name)] panel which are read in from a file are displayed in the display panel.
Section 2 System Overview 2-5-3 Drag and Drop Function Execution Icon This software uses the drag and drop method of selecting image files and observation methods (dye color name or transmitted light). With this intuitive method, selection requires only picking up [Item to Select (image file or obsen/ation method)] with the mouse and dragging it to the setting location, where it is dropped. * ^ " • . • : . ^ - \ • : : • • • • ' . • , : ' . . • .Oi1:FITC!:."=.---:'':-; S-'-.--. ,;CH2:Rhodamin... ;,•.
Section 2 System Overview 2-6 System Setup This system must be set up by a special technician. To maintain the perfonnance and for safety, never disassemble or re-adjust this system. 2-6-1 Power Consumption As shown below, some of the units consume considerable power; therefore, check the capacity of the power outlet before connecting this system. Electromagnetic Control unit IHard copy unit disk Microscope 3A Max. 3A Max. Personal computer j 0,3A Max.
Section 2 System Overview 2 - 6 - 2 Turning o n the Power Supply Tum on the power supply of each unit. When only reading data, the laser power supply and illumination power supply are not necessary and need not be tumed on. To obtain a stable laser output, tum on the laser power supply and allow the system to warm up for 10 min. or more before using. Once the illumination power supply (mercury vapor lamp power supply) has been tumed off, do not turn it on again for at least 10 min.
Section 2 System Overview • Tuming on the power controller power supply (1) Tum on the main switch. For details, refer to the Power Controller SSI-9501 instruction manual. (2) Tuming on the switches If all of the switches are tumed on, the power for the entire system can be tumed on and off with the main switch as described in item (1)The units are connected as shown below.
Section 2 System Overview Tuming on the control unit power supply (1) Turn on the power supply switch. This unit is connected to the power controller, so the power switch can be left turned on. • Tuming on the electromagnetic disk power supply (Please read the electromagnetic disk instruction manual.) This unit is connected to the power controller, so the power switch can be left turned on. • Tuming on the microscope power supply (Please read the instruction manual of the microscope used.
Section 2 System Overview 2-7 System Operational Procedure Overview A flow chart is used here to explain the operational procedures so that operation of the entire system can be more easily understood. For reference, the relevant manual sections (f (( )) are indicated at the right side of the flow chart for each procedure. j) and numbers Please refer to 2-7-1 Fluorescent Light Observation Procedure and 2-7-2 Transmitted Light Observation Procedure.
Section 2 System Overview 2-7-1 Fluorescent Light Observation Procedure Start the system. Prepare the system (tum on the power supply). Introduction to Fluoview (2-6) i Replace with a high transmt^lifQn rate ND filter (100%, 50%, 20%»6%). Hardware -, (3-1-6) . . , Start the Ftyn( 2-1-1) Read in an image. Operation (3-1) Look through the eiej^Hnti J^ns and focus on the speclmea ISelect the l i ^ p f ^ f o r 100% for binocular tube andfeiCttSidnthe specimen.
Section 2 System Overview 2-7-2 Transmitted Light Observation Procedure start the system. Prepare the system (tum on the power supply). Introduction to Fluoview (2-6) ^Replace with a high transmis^i;^ rate^ ND filter (100%, 50%, 20%,^5%). Hardware ' (3-2-6) start the fluoview softvyrare. Operatton (2-1-1) Read in an image. Operation (3-1) . Look thraugti the eyd(jto0^l0ns and focus on ttie specimen. " Select the ^ ; path for 100% for ^ binocular tube andfocuson the specimen.
Section 2 System Overview 2-8 Identifying Images From Different Methods of Observation Fluoview displays various image icons which can be used to identify the method of obsen/ation used to read in images.
This instmction manual describes functions, specifications, assembly (connections) and adjustment of the hardware. Before reading this instmction manual, please thoroughly review the preceding manual "INTRODUCTION FLUOVIEW" for an outline of the system.
LASER SAFETY PRECAUTION Units indicated as "OPERATOR SERVICE" should be removed only by a person who has had laser safety training, and only after tuming off the laser unit. This should not be done by any untrained persons. Removing any of the following units is considered to be OPERATOR SERVICE : Cube turret Objective revolving nosepiece Objective lens DIC slider or dummy slider Transmitted light lamphousing and / or lamp socket Detection mode selector slider ( Scan unit) See 2-1, 2-7, 2-10.
1 1 STANDARD CONFIGURATIONS STANDARD CONFIGURATIONS 1-1 BX Upright Microscope System Configuration bfesi&;SftM!3escnpbonMH?#g^ ilBX5oy^ti§ S^^^ii 1 Scan Unit FVX-SU 1 Excitation Cube FVX-DM488 1 Filter Dual Wavelength Excitation Cube FVX-BA565IF 1 Banier Filter FVX-BA550RIF Barrier Filter FVX-BA585IF Kr and Ar Laser Line Filter Polarizing Filter for Upright Microscope FVX-LLF-KR Control Unit FVX-CU Pupil Lens FVX-PL-IBX50 Laser Tube FVX-LT Extension Unit FVX-EXTU Stand for BX Rub
CONTENTS 1 STANDARD CONFIGURATIONS t£t 1-1 BX Upright Microscope System Configuration 1-1 1-2 IX Inverted Microscope System Configuration 1-2 1-3 Optional Accessories 1-3 2 MAIN UNITS AND DESCRIPTION OF CONTROLS 2£L 2-1 Scan Unit 2-1 2-2 Microscope Frame 2-7 2-3 Transmitted Light Detector (Optional) 3 PREPARATIONS FOR OBSERVATION 3-1 Fluorescence Observation 2-13 Z=l 3-1 3-1-1 Bringing the Specimen into Focus 3-1 3-1 -2 Selecting the LSM Light Path 3-3 3-1-3 Selecting the Pinhole 3-5
1 STANDARD CONFIGURATIONS 1-2 IX Inverted Microscope System Configuration ^nimsii^ «Siiii^«»m m m ^ .
1 STANDARD CONFIGURATIONS 1-3 Optional Accessories Green HeNe Laser Unit FVX-LU-HEG Used In combination with the Ar laser. Suitable for TRITC and PI obsen/ation. 543 nm excitation wavelength. Can not be combined witii the Kr laser. Dual Wavelength Excitation ~ Cube FVX-DM488/543 Dual wavelength excitation cube for Ar laser and HeNe green laser. Banier Filter FVX-BA530RIF Short-pass filter for blocking ttie 543 nm wavelength beam emitted by the HeNe green laser.
2 MAIN UNITS AND DESCRIPTION OF CONTROLS 2 MAIN UNITS AND DESCRIPTION OF CONTROLS Also referto Section 2-4, SYSTEM LAYOUT in the "INTRODUCTION TO FLUOVIEW manual. 2-1 Scan Unit "OPERATOR SERVICE" (2) Detection mode _ (D Pinhole tun^t selector slider (5) Filter slider cover screws USBt I I I E K I T r 0 o(E)« :iECTION WOE Ofl OC H. IR H. H. TR R.
2 MAIN UNITS AND DESCRIPTION OF CONTROLS In general, selecting a smaller pinhole than the pinhole size recommended for a given objective will not increase the axial resolution. However, in some cases the lateral resolution may increase. The effect differs with the refractive index of the specimen and the dispersion of the light The following table shows-the recommended pinhole number for a given objective. Objective \ i ^ NA Pinhole No. PLAPO 40X 0.95 2 PLAPO 60XO 1.40 2 PLAPO 100XO 1.
2 MAIN UNITS AND DESCRIPTION OF CONTROLS (2) Detection mode selector slider Slider for selecting either two fluorescence signals or one fluorescence signal + transmitted light 3 settings. This slider is placed at the A-section shown in Fig, (I). • Slide'r at the pushed-in position Select this mode when observing a transmitted light and / or a fluorescence with emission wavelength shorter than 570nm, such as FITC, GFP, DiO, etc. As shown in Fig.
2 MAIN UNITS AND DESCRIPTION OF CONTROLS Laser Slider at pushed-in position CHI CH2 FL TR Fig. ( I ) Middle position Fig.(n) Slider at pulled-out position Fig.
2 MAIN UNITS AND DESCRIPTION OF CONTROLS (3) Bamer filter slider Up to two filters can be placed for both CHI and CH2. At the pushed-in position, the filter is engaged into the light path. At the pulled-out position, it is disengaged. * If erroneously placed at the middle position, no image will appear. Always stop at the correct position. (4) ND filter turret There are five positions. Transmission ratio: 0, 6, 20, 50, and 100%.
2 MAIN UNITS AND DESCRIPTION OF CONTROLS • FITC emission tails longer than 570 nm. Accordingly, FITC fluorescence may be detected on the CH2 where it overlaps the Pl fluorescence. (See the figure below.) The problem may be remedied by cutting the FITC by adjusting the OFFSET on the operation panel ([Acquire] panel). If not, It will be necessary to balance the excitation light by reducing the intensity of the 488 nm excitation light used for exciting the FfFC.
2 MAIN UNITS AND DESCRIPTION OF CONTROLS 2-2 Microscope Frame The illustration below shows the main controls of the microscope frame. The stage, revolving nosepiece, etc., may differ from those illustrated. For details on operation of the microscope frame, refer to the instruction manual for tiie microscope frame.
2 MAIN UNITS AND DESCRIPTION OF CONTROLS (1) Light path selector • When the knob is pushed-in, visual observation is possible. • When the knob is pulled-out laser microscopy is possible. (2) Cube turret • Engage the designated cube for visual fluorescence observation. • When used as a laser microscope and for visual transrnitted light observation, operate the turret to place the index at the [ { Q )] position. (Set the cube turret so tiiat no cube is engaged.
2 MAIN UNITS AND DESCRIPTION OF CONTROLS (2) Universal condenser For Nomarski observation, engage the Nomarski prism (optional) suitable for the objective in use. (Similar for both visual and laser Nomarski observation.) Note that tiie polarizer should also be engaged in the case of laser Nomarski obsen/ation. (Note that the analyzer U-AN should be removed from Uie light path in tiie case of laser Nomarski observation.
2 MAIN UNITS AND DESCRIPTION OF CONTROLS IX Confiquration "OPERATOR SERVICE" • transmitted light lamphousing • lamp socket (6) Filter "OPERATOR SERVICE" objective lens "OPERATOR SERVICE" (4) DIC prism U-DICT(dummy slider) (optional) (1) Light path selector "OPERATOR SERVICE" (2) Cube tunet (7) Magnification selector knob (3) Analyzer IX-AN 2-10
2 MAIN UNITS AND DESCRIPTION OF CONTROLS (1) Light path selector • When tiie knob is set at the ^ [ position, visual observation is possible. • Set the knob at the ( SP ] position for laser microscope use. (2) Cube turret • Engage the designated cube for visual fluorescence observation. • When used as a laser microscope and for visual transmitted light observation, operate tiie turret to place the index at the [ [ O )] position. (Set the cube tun'et so that no cube is engaged.
2 MAIN UNITS AND DESCRIPTION OF CONTROLS (2) Condenser For Nomarski obsen/ation, engage the Nomarski prism (optional) conesponding to the objective in use. (This applies to both visual and laser Nomarski obsen/ation.) Note that the polarizer should also be engaged for Nomarski obsen/ation. (Note that the analyzer IX-AN should be disengaged for laser Nomarski observation.) (3) Magnification selector knob For LSM use, always select IX (knob pushed-in). The 1.5X setting cannot be used.
2 MAIN UNITS AND DESCRIPTION OF CONTROLS 2-3 Transmitted Light Detector (Optional) FVX-TD-BX (for BX50, BXWI) (1) Light paUi selector • When the knob is pushed-in, laser transmitted light observation is possible. • When the knob is pulled-out visual transmitted light observation is possible. (D Light path selector FVX-TD-IX (for IX) • When the knob is pushed-in, laser transmitted light observation is possible. • When the knob is pulled-out, visual transmitted light observation is possible.
3 PREPARATIONS FOR OBSERVATION 3 PREPARATIONS FOR OBSERVATION This section explains the order of procedures for specimen obsen/ation. 3-1 Fluorescence Observation 3-1-1 Bringing the Specimen into Focus 3-1-1-1 BX50, BXWI Configuration 1. Push in the light path selector knob (1) on tiie binocular observation tube to select Uie 100% at binocular eyepieces setting. 2. Operate the cube turret to engage the cube corresponding to the specimen fluorochrome. 3.
3 PREPARATIONS FOR OBSERVATION 3-1-1-2 IX Configuration 1. Turn the light paUi selector dial (1) on ttie right side of Uie microscope to the ^ position. While looking through the eyepieces, bring the specimen into focus. Make sure to adjust the diopter adjustment ring of Uie eyepiece. (Refer to the 1X50/70 instiuction manual.) 2. Operate the cube tun-et to engage the cube corresponding to the specimen fluorochrome. 3. While looking through the eyepieces, bring the specimen into focus.
3 PREPARATIONS FOR OBSERVATION 3-1-2 Selecting the LSM Light Path 3-1-2-1 BX50, BXWI Configuration 1. Pull out the light path selector (1) on the trinocular obsen/ation tube to the stop position. 2. Operate the cube turret (2) on the vertical illuminator to align the index with the [( Q ) ] position. 3. If the analyzer U-AN (3) is mounted, disengage it by pulling it out to the pulled-out clickstop.
3 PREPARATIONS FOR OBSERVATION 3-1-2-2 IX Configuration 1. Tum the light path selector (1) to the [[ SP J ] position. 2. Set the magnification selector knob (7) to lx. 3. Rotate the cube turret to select [ ( Q ]]. 4. If the analyzer IX-AN (3) is mounted, disengage it by pulling it out to the far clickstop. (Leaving the U-DICT engaged during laser fluorescence observation will degrade tiie image quality somewhat.
3 PREPARATIONS FOR OBSERVATION 3-1-3 Selecting the Pinhole Operate the pinhole tun-et (1) to select ttie pinhole corresponding to tiie pinhole number indicated for each objective on tiie control panel. (Refer to Section 2-1, Scan Unit) 3-1-4 Selecting the Detection Mode Set ttie detection mode selector slider to the designated position in accordance with Uie fluochrome ofthe specimen to be observed.
3 PREPARATIONS FOR OBSERVATION (2) [}etection mode selector slider (5) FIKer slider cover screws (1) Pinhole turret ~B| UBBt IMTEI6ITY 0 9(S)9 OLYMPUS FLUOVIEW 0 Jt (4) ND filter turret (3) Barrier filter slider (7) Laser line filter turret (only for Kr and Ar laser combination) 3-1-6 Selecting the ND Filter Operate tiie NDfilterturret (4) to select the appropriate NDfilter.Select the NDfilterin accordance with the specimen brightness and level of photobleaching.
3 PREPARATIONS FOR OBSERVATION 3-2 Transmitted Observation The procedures for transmitted observation are similar to those forfluorescenceobservation. Also refer to Section 3-1, Fluorescence obsen/ation. 3-2-1 Selecting the Transmitted Light Detector Light Path (Visual Setting) 3-2-1-1 BX50, BXWI Configuration Pull out the light path selector (1). (D Light path selector 3-2-1-2 IX Configuration Pull out the light path selector (1).
3 PREPARATIONS FOR OBSERVATION 3-2-3 Selecting the LSM Light Path 3-2-3-1 BX50, BXWI Configuration 1. 2. Pull out Uie light path selector (1) on Uie trinocular observation tube to the pulled-out clickstop. Operate the cube turret (2) on the vertical illuminator to align the index with Uie [Col] 3. 4. 5. If the analyzer U-AN (3) is mounted, disengage it by pulling it out to the pulled-out clickstop. (It makes no difference if the DIC prism U-DICT (4) remains engaged.
3 PREPARATIONS FOR OBSERVATION 3-2-3-2 IX Configuration 1. Tum the light path selector (1) to select the [[ sP W setting. 2. Set the magnification selector knob (7) to IX. 3. Operate the cube turret4o select the [ ( Q J position. 4. If the analyzer U-AN (3) is mounted, disengage it by pulling it out to the far clickstop. (It makes no difference if the DIC prism U-DICT (4) remains engaged into the light paUi.) 5. Remove all the filters from the light path. 6.
3 PREPARATIONS FOR OBSERVATION 3-2-4 Selecting the Detection Mode When the detection mode selector slider (2) is: • Pushed in, transmitted light is detected by CH2. • Pulled out, transmitted light is 'detected by CHI. Consequently, when only transmitted observation is performed, the detection mode selector slider (2) may be set at either position.
3 PREPARATIONS FOR OBSERVATION 3-2-5 Disengaging the Barrier Filter Remove all the barrierfiltersfrom the channel to detect transmitted light. (The banier filter slider (3) should be at ttie pulled-out position.) If in doubt about tiie setting, refer to Section 4-1-2, Adjusting the Scan Unit in the Operation Manual and follow ttie prompts of ttie [Microscope Configuration] window. 3-2-6 Selecting the ND Filter Operate the NDfilterturret (4) to engage a suitable ND filter.
4 SPECTRAL CHARACTERISTICS OF FILTERS 4 SPECTRAL CHARACTERISTICS OF FILTERS Barrier filters BA585IF BA510IF Representative filter combinations for Ar laser use BA565IF ( EDM570 is a dichroic mirror for splitting fluorescence light into CH1/CH2. DM488 is dichroic mirror for excitation.
4 SPECTRAL CHARACTERISTICS OF FILTERS • Representative filter combination for the Kr and Ar laser combination DM488/568 \ , BA510IF 100 EDM570 is a dichroic min-or for splitting fluorescence light into CH1/CH2. DM488/568 is dichroic 50 - mirror for double excitation for use with Uie KrAr laser.
5 SPECIFICATIONS 5 SPECIFICATIONS -:^^M^:r^M^^M^'^:^ir^'$^ Laser unit Scan unit Control unit '^^i}:k;^:/i:l-^r!-f''!'^^U%^ Ar laser 5 mW output, random or linear polarization. Air cooled Argon ion laser (488 nm) Scan unit connection: Single mode optical fiber (3 m) Power requirements: 100V 10A (MAX) Kr laser 15 mW output, linear polarization.
5 SPECIFICATIONS 5 SPECIFICATIONS ;:«§p^p5if^6^^ipjKfi|ps^?i^^ Laser unit Scan unit Control unit •li^^^li^fiiPi^a«iii« Ar laser 5 mW output, random or linear polarizaition. Air cooled Argon ion laser (488 nm) Scan unit connection: Single mode optical fiber (3 m) Power requirements: 100V 10A (M/VX) Kr laser 15 mW output, linear polarization.
5 SPECIFICATIONS ?^-ri^^.
5 Operating environment SPECIFICATIONS Indoor use. Altitude: Max. 2000m Ambient temperature: 5°C to 4 0 t (41* F to 109* F) 10°C t o 3 5 t (Performance guarantees) Maximum relative humidity 80% for temperatures up to 31°C (88' F) decreasing lineariy tiirough 70% at 3 4 t (93' F), 60% at 3 r t ( 9 9 * F). to 50% relative humidity at 40°C (109" F) Main supply voltage fluctuations not to exceed ±10% of the nominal voltage.
II mil inie^QQinij Q ^' Concerning This section explains countermeasures to be taken in the event that a problem occurs. Please read this section and try the countermeasures before contacting Olympus. If the problem persists, then contact Olympus.
Contents Section 1 Problem Countermeasures 1-1
Section 1 Problem Countermeasures Section 1 Problem Countermeasures Due to Uie usage, the functions of the system sometimes are not fully effective, although there has been no failure; tiierefore, when a problem occurs, refer to the following chart and apply the appropriate countermeasures. If the same phenomenon continues to occur after the countermeasures have been taken, contact your local Olympus dealer. fea^AJi^,SyniptOmff;;^ia^ 1. No fluorescent light image. The laser is not oscillating. ^:j;^.^.
Section 1 Problem Countermeasures ^ ^ K ^ ^ ^ ^ S :^Kl^aus^i^Si^ g^^^'JCountemrieasureliJ^sg ?Manual!Reference^ 1. No fluorescent light image. The photomultiplier of the Check the [Chi] and [Ch2] Hardware, 2-1. channel set with the check boxes of the [Acquire] Operation, 4-1-3-3. detection mode setting knob panel. is not set. 2. No transmitted light image. The transmitted light Con-ectly set ttie transmitted Hardware, 3-2-3. detector is not correctiy set.
Section 1 Problem Countermeasures ^ ^ ^ « 6. Flare is visible. ^ •^m^amm ftmmQounleTmeaswe"MMi ;iManuai,iReference^' The cover glass ttiickness is Use a cover glass witti a not suitable. thickness of 0.17mm. — The specimen is overstained. 7. Circular flares appear in the center of the image. 8. Blun-ing is visible. Use an appropriate dye color Operation, 4-1-3-10. or increase the offset value.
Section 1 Problem Countermeasures teg5l3i?^tSymptomMs4;!Si;#Si The Z stage (Z revolver) 13. The scale of tiie [Z motor is not being excited. Stage] in the [Acquire] panel is not woridng. 14. The fine movement handle of the microscope will not move or moves with difficulty. The Z stage (Z revolver) motor is being excited. 15. Images cannot be saved The disk has not been formatted (in the case of a to disk. floppy disk or photoelectric disk). The photomagnetic disk is not recognized.
