Page 1 HistoCore PEGASUS Tissue Processor Instructions for Use English Order No.: 14 0488 81101 - Revision J Always keep this manual with the instrument. Read carefully before working with the instrument. Version 1.6, Revision J - 11.2023...
Page 3 – including any electronic systems and media – requires express prior permission in writing by Leica Biosystems Nussloch GmbH. For the instrument serial number and year of manufacture, please refer to the nameplate on the back of the instrument.
Page 4: Table Of Contents
Table of Contents Important Information ....................8 Naming conventions ........................8 Symbols and their meanings ....................... 8 Instrument type ........................12 Intended purpose ........................12 1.5 Qualification of personnel ......................12 Statement for user data security and privacy ................12 Safety ........................13 Safety notes ..........................13 Warnings ..........................14 2.2.1 Markings on the instrument itself ....................14 2.2.2 Transport and installation......................14 2.2.3 Operating the instrument ......................16...
Page 5 7.1.6 Reagent compatibility ......................109 Managing reagent types ......................109 7.2.1 Pre-defined reagents .......................110 7.2.2 Editing active reagent types .....................110 7.2.3 Adding, hiding, and deleting reagents ..................112 Managing reagent stations ......................114 7.3.1 Reagent stations screen ......................115 7.3.2 Setting reagent station properties ....................116 HistoCore PEGASUS...
Page 6 Table of Contents Replacing reagents .........................119 7.4.1 Remote fill/drain screen ......................119 7.4.2 Remote fill/drain connections ....................120 7.4.3 Replacing reagent – remote fill and drain .................121 7.4.4 Replacing reagent – manually ....................124 7.4.5 Replacing paraffin ........................125 7.4.6 Filling and draining retorts .......................127 Ancillary Settings & Operations ................. 128 Reagents menu ........................128 8.1.1 Manual operations ........................128 Administration menu .......................129 8.2.1 User management ........................130 8.2.2 Reports ..........................131...
Page 7 11.2.7 Staining artifact ........................175 11.2.8 Block artifact ..........................176 11.2.9 Reprocessing recommendations ....................177 11.3 Power failure ..........................180 11.4 Crash or freeze of master software ..................180 Ordering Information ....................181 A1. Decontamination Confirmation .................. 182 A2. Warranty and Service ....................183 HistoCore PEGASUS...
Page 8: Important Information
Important Information Important Information Naming conventions Note • The full name of the device is HistoCore PEGASUS tissue processor. The device is called HistoCore PEGASUS to ensure that the Instructions for Use are well legible. Symbols and their meanings Symbol: Title of the symbol: Danger Description: Indicates an imminently hazardous situation which, if not avoided, will result in death or serious injuries.
Page 9 The UK Responsible Person acts on behalf of the non-UK manufacturer to carry out specified tasks in relation to the manufacturer’s obligations. Symbol: Title of the symbol: WEEE Symbol Description: The WEEE symbol, indicating separate collection for WEEE – Waste of electrical and electronic equipment, consists of the crossed-out wheeled bin (§ 7 ElektroG). HistoCore PEGASUS...
Page 10 Important Information Symbol: Title of the symbol: China ROHS Description: Environmental protection symbol of the China RoHS directive. The number in the symbol indicates the "Environment-friendly Use Period" of the product in years. The symbol is used if a substance restricted in China is used in excess of the maximum permitted limit.
Page 11 With a pitch of 60° or more, the blue quartz sand flows into the arrow-shaped indicator window and sticks there permanently. Improper handling of the shipment is immediately detectable and can be proven definitively. Symbol: Title of the symbol: CSA symbol Description: This product fulfills the requirements of the CAN/ CSA-C22.2 No. 61010. HistoCore PEGASUS...
Page 12: Instrument Type
Instructions for Use. Qualification of personnel • The HistoCore PEGASUS must be operated by trained laboratory personnel only. The instrument is intended for professional use only. • All laboratory personnel designated to operate this instrument must read these Instructions for Use carefully and must be familiar with all technical features of the instrument before attempting to operate it.
Page 13: Safety
• The protective devices located on the instrument and the accessories must not be removed or modified. Only qualified service personnel authorized by Leica Biosystems may repair the instrument and access its internal components. • If the instrument is to be returned to Leica Biosystems for repair, it must be cleaned and decontaminated in the appropriate manner (→ p. 182 – A1. Decontamination Confirmation).
Page 14: Warnings
Safety The electromagnetic environment should be evaluated prior to the operation of the instrument. Do not use this instrument in close proximity to sources of strong electromagnetic radiation (e.g. unshielded intentional RF sources), as these can interfere with the proper operation. Warnings The safety devices installed in this instrument by the manufacturer only constitute the basis for accident prevention.
Page 15 • Only process patient tissue samples for diagnostic purposes, after all the above checks have been performed successfully. If any of the previous steps failed, do not operate the instrument and contact your local Leica Biosystems Service Representative. HistoCore PEGASUS...
Page 16: Operating The Instrument
Safety Warning Insufficient or no exhaust system/Wrong connection to the exhaust system People injury due to leaked vapor or excessive fumes • Never run the instrument without the active carbon filter or an external exhaust system, as this will release potentially dangerous fumes into the laboratory. • It is recommended to use an external exhaust system. • Even if the instrument is connected to an external exhaust device, the active carbon filter provided must remain in use. Warning The instrument is plugged into a power board with other instruments. The instrument may work with unstable current/power, which leads to sample damage.
Page 17 • Do not set the RMS to N/A. Use RMS to monitor reagent consumption status. • The RMS settings must be validated by the user before running the protocol, i.e. tissue processing with patient tissue for diagnostics, according to the local or regional accreditation requirements. HistoCore PEGASUS...
Page 18 Safety Warning Open retort while the instrument is processing/after pausing/after the protocol ends, the user gets contact with the hot surface of lid/baskets/fluids/hazardous reagents/vapor from the retort. Open the retort and the protocol cannot continue. People injury/Sample damage/Impaired samples • Hot surface. Wear chemical protective clothing, protective goggles, respirator, rubber gloves, and all other necessary personal protective equipment during operation.
Page 19 Ignore the information provided by the instrument. Update reagent/station parameters but not replace the reagent. Do not update the reagent status on GUI from Empty to Full after filling the bottle. Or set the status to Full before filling the bottle, but not filling the bottle or underfilling the bottle. Sample damage caused by dirty or expired reagent, or insufficient reagent. Delay of diagnosis. • Always change reagents when prompted. • Always update station details correctly. • Never update the details without replacing the reagent. HistoCore PEGASUS...
Page 20 Safety Warning Fill the reagent from an unstable container. People injury - The fill/drain functions include a strong purge which may cause an unstable container to tip over and spill. The user may slip on the spilled reagent on the ground. Environment contamination. • Always ensure that you fill from or drain to a large stable container. The container must also be of sufficient volume to easily accommodate all of the drained fluid. If you use a small container you must support the container and tube during the fill or drain. • Always use the tube supplied with the instrument. •...
Page 21 • If you inadvertently load unprocessed samples into a retort prior to running a cleaning protocol, remove the samples and attempt to load a processing protocol before loading the cleaning protocol. The purge before the cleaning run will be skipped. HistoCore PEGASUS...
Page 22 Safety Warning Run cleaning protocol to clean additional materials other than baskets. Impaired samples - Cleaning reagents degenerate at a faster rate. If the cycle purity threshold is not lowered, the low purity cleaning reagents leads to low-quality cleaning. • Do not clean other materials using cleaning protocol. Warning Add fixed samples to a running protocol.
Page 23: Handling Reagents
• The DC overcurrent conditions are protected against by the separated heating power fuses. Overheating protection The heat limiters and software overheat error handling mechanism protect the instrument against the abnormally high temperature. Over vacuum protection The vacuum system is not capable of generating a dangerous vacuum condition. HistoCore PEGASUS...
Page 24: Instrument Components And Specifications
Instrument Components and Specifications Instrument Components and Specifications Overview 3.1.1 Instrument components Fig. 1 1. Display 7. Drip tray 2. Four paraffin baths 8. Condensate bottle 3. Two retorts 9. Active carbon filter (behind the cover) 4. Remote fill/drain port (reagent) (behind the cover) 10. Mains power switch 5. Remote drain port (paraffin) (behind the cover) 11. Tool box 6. Reagent bottles 12. HistoCore I-Scan (Optional) 13.
Page 25: Rear Panel
6. Exhaust interface (connect to an external exhaust device) 3. Circuit breaker 7. Nameplate 4. Mains power inlet Main features of the instrument The HistoCore PEGASUS (hereafter PEGASUS) is a dual-retort tissue processor that offers fast, high- quality tissue processing for histology laboratories. With reagent capacity sufficient to run the two retorts independently, it allows high-degree scheduling flexibility. Intelligent software lets you configure the instrument to suit your laboratory’s workflows while providing checks and restrictions that help you avoid mistakes that could damage tissue.
Page 26: Technical Data
Instrument Components and Specifications Pre-defined validated protocols give you the flexibility to process different tissue types with consistent quality. The magnetic stirrer technology is designed to optimize paraffin infiltration performance, helping to reduce processing time and enhancing reagent exchange within the tissue cells. Backlighting allows you to check the reagent level easily. When the reagent bottle is not connected correctly, the backlight will turn off. Filling and draining reagents and paraffin can be done with the remote fill/drain function which minimizes exposure to reagents and hot paraffin. Reagent bottles with an Anti-spillage design which avoids reagent spill during reagent transfer. Speed up the paraffin melting time to 3.5 hours with HistoCore PEGASUS Parablocks, which perfectly matches the paraffin chamber paraffin fill volumes without topping off, and the elimination of pellet spills.
Page 27 17 reagent bottles, 1 condensate bottle Maximum bottle volume Max. 3.88 L Paraffin containers Number of paraffin baths Capacity Max. 3.9L per paraffin bath Melting time Max. 7 hours an ambient temperature of 23°C to 25°C (set the paraffin type as "Recommended") Temperature settings range • Paraffin Melting Point setting: 50°C to 65°C • Standby temperature setting: Paraffin Melting Point +2°C and max. 71°C HistoCore PEGASUS...
Page 28 Instrument Components and Specifications Retort Capacity Max. 2 x 200 cassettes Temperature (paraffin) Paraffin temperature selectable from 58°C to 77°C. Temperature (processing reagents) Ambient temperature or 35°C to 65°C Temperature (cleaning reagents) 35°C to 85°C Filling vacuum -60 kPa Draining pressure 45 kPa Other specifications Approved Approved Reagent Management System (RMS) •...
Page 29 Instrument Components and Specifications Other specifications Capacities • 20 freely configurable protocols. 4 pre-defined protocols. Xylene pre-defined protocols consist of up to 10 reagents and 3 paraffin processing steps. Xylene-free pre-defined protocols consist of up to 8 regents and 3 paraffin processing steps. --Time per protocol step: 0 - 5999 minutes. --Delay time: max. 1000 hours. • Up to 400 cassettes can be processed simultaneously. • 1 pre-defined cleaning protocol for the retort. HistoCore PEGASUS...
Page 30: Setting Up The Instrument
• Use only the provided power cords intended for the local power supply. Standard delivery - packing list Quantity Part description Order No. HistoCore PEGASUS basic instrument 14 0488 58006 Basket hook 14 0488 58177 Cassette basket 14 0488 58160...
Page 31 14 0488 81200) If the supplied local power cord is defective or lost, please contact your local Leica representative. Note • Please check all delivered parts against the packing list and against your order to verify whether the delivery is complete. Should you find any discrepancies, please contact your Leica Biosystems sales office without delay. HistoCore PEGASUS...
Page 32: Unpacking And Installation
Setting up the Instrument Unpacking and installation 4.3.1 Unpacking Instructions Warning Accessories/components may become loose/damaged during transportation Sample damage • The packaging has two indicators, ShockDot Impact Indicator (→ Fig. 3-1) and Tilt Indicator (→ Fig. 3-2), that indicate improper transport. When the instrument is delivered, check this first. If one of the indicators is triggered, the package was not handled as prescribed. In this case, please mark the shipping documents accordingly and check the shipment for damage.
Page 33 9. Hook these rails on the left and right with the panel into the groove of the pallet so that they are flush with the wooden board that holds the instrument. 10. Now release the brake lever on the two front transport castors so that the instrument can be moved. Fig. 6 HistoCore PEGASUS...
Page 34 Setting up the Instrument Warning The instrument tilts while moving during the first installation. People injury • At least two persons are required to hold the instrument when it is rolled off the pallet via the ramp. 11. When rolling the instrument down from the pallet, support it by the outer top corners using both hands.
Page 35: Installing The Display
• Even if the instrument is connected to an external exhaust device, the active carbon filter provided must remain in use. The active carbon filter absorbs reagent fumes, thus preventing them from entering the laboratory atmosphere. To ensure that the active carbon filter operates effectively, it must be replaced periodically. Supervisors can set a replacement threshold by tapping Carbon filter threshold on the Settings > Instrument settings screen. To insert/replace the active carbon filter, open the remote fill/drain cover (→ Fig. 9-1). Then push the active carbon filter (→ Fig. 9-2) in as far as it will go. The date on which the active carbon filter has been inserted can be marked on the label on the front side. HistoCore PEGASUS...
Page 36: External Exhaust System
Setting up the Instrument Fig. 9 4.3.4 External exhaust system Warning Insufficient or no exhaust system/Wrong connection to the exhaust system People injury due to leaked vapor or excessive fumes • Never run the instrument without the active carbon filter or an external exhaust system, as this will release potentially dangerous fumes into the laboratory. • It is recommended to use an external exhaust system. • Even if the instrument is connected to an external exhaust device, the active carbon filter provided must remain in use.
Page 37: Basic Instrument/Hardware
This releases the e-lock. The retort lid can be opened. Then remove the straight screwdriver and insert the black cover back to the place. HistoCore PEGASUS...
Page 38 Setting up the Instrument Fig. 12 Warning Open retort while the instrument is processing/after pausing/after the protocol ends, the user gets contact with the hot surface of lid/baskets/fluids/hazardous reagents/vapor from the retort. Open the retort and the protocol cannot continue. People injury/Sample damage/Impaired samples • Hot surface. Wear chemical protective clothing, protective goggles, respirator, rubber gloves, and all other necessary personal protective equipment during operation.
Page 39 It serves a safety function. The sensors may occasionally be affected by a build-up of condensation or deposited material. Always ensure the sensors are kept clean as part of regular retort cleaning(→ p. 148 – 9.3.1 Daily tasks). HistoCore PEGASUS...
Page 40: Cassette Baskets
Setting up the Instrument Magnetic stirrer Fig. 15 Each retort has a magnetic stirrer (→ Fig. 15-1) that stirs the reagent or paraffin to ensure a consistent temperature and good tissue penetration. The stirrer is driven by a magnetically-coupled external motor. The stirrer can be turned on or off for each protocol step and is easily removed for cleaning (→...
Page 41 (→ p. 142 – 8.3.4 Tracking options (optional)). After scanning the barcode and taking the basket image, you can place the basket in the retort. It is recommended to use Leica Biosystems pre-defined/validated consumables (baskets and cassettes). HistoCore PEGASUS...
Page 42: Paraffin Baths
Setting up the Instrument 4.4.3 Paraffin baths Fig. 18 Four paraffin baths are located at the top rear of the instrument accessed by two paraffin bath lids (→ Fig. 18-1). Each bath acts independently, holding enough paraffin to fill a retort. Use the paraffin bath lid handles to open the paraffin bath lids. Be careful when opening the paraffin bath lids. If needed, rotate the display to give easy access to the paraffin bath (→ Fig. 23). Always use the paraffin bath handles to close the lids, ensuring the lids are properly latched. View the current temperature of each paraffin station by going to Reagents > Stations > Paraffin chambers. Drain used paraffin with commands on the Reagents > Remote fill/drain screen. Directly fill the paraffin baths with molten or solid paraffin (→ p. 125 – 7.4.5 Replacing paraffin). Version 1.6, Revision J...
Page 43: Reagent Cabinet
The reagent bottle at each station holds sufficient reagent to fill a single retort. The reagent bottles are translucent and have color-coded labels. They also have markings on the front showing the minimum level (→ Fig. 20-1) for a run and the maximum fill level (→ Fig. 20-2). HistoCore PEGASUS...
Page 44 Setting up the Instrument Fig. 20 To remove a reagent bottle, pull the reagent bottle out. Push back in position to return. You will hear a click sound and feel the bottle fitting into its connection point on the back of the cabinet. You can replace the reagent in a reagent bottle either: • manually, after having removed the reagent bottle from the instrument, or •...
Page 45: Drip Tray
The round labels (→ Fig. 21-2) are applied to the reagent bottle caps. The surface on the reagent bottles must be clean and dry; then apply the label and press on firmly – especially at the edges. The labels are resistant to the reagents used in the instrument. 4.4.5 Drip tray Fig. 22 HistoCore PEGASUS...
Page 46: Display
Setting up the Instrument The drip tray is located along with other accessories in the packing box. Push the drip tray (→ Fig. 22-1) under the reagent cabinet. The drip tray collects reagents that have overflowed or spilled so that no pollution can form in or under the instrument. The volume is not less than 3.88L. The drip tray must be regularly checked for signs of reagents that have leaked. To do so, pull the tray out by the handle and empty it if necessary.
Page 47: Histocore I-Scan (Optional)
A tick is shown after the instrument receives the image successfully, or a warning mark is shown. Hand shake during focusing and image capturing will cause the picture unclear. It takes about 2 seconds for the camera to focus and capture an image. HistoCore PEGASUS...
Page 48: Usb Ports
Setting up the Instrument Button Name Function Tick button Confirm the entry and enter the next filling box. If the recognition of a barcode fails, please try to scan it again. The recommended scanning distance is as follows. • EAN-13 50mm-200mm (13mil) • Code39 40mm-90mm (5mil 10 Byte) •...
Page 49: Alarm Connections
(connectors inserted while alarms not connected) should never appear on an instrument. Alarm connector specifications The load connected to the alarm connector must not exceed the following specifications. • Maximum voltage: 30 V DC • Maximum current: 1 A (resistive load) • Maximum current: 0.5 A (inductive load) HistoCore PEGASUS...
Page 50: Connecting An Uninterruptible Power Supply (Ups)
Setting up the Instrument Alarm connector pins Each alarm connector has three pins. Remote alarm RA.NC (→ Fig. 26-1) -- Normally closed; RA.COM (→ Fig. 26-2)-- Common; RA.NO (→ Fig. 26-3) Normally open Local alarm LA.NC (→ Fig. 26-4), -- Normally closed; LA.COM (→ Fig. 26-5) -- Common; LA.NO (→ Fig. 26-6) Normally open Pin schematic during normal operation When the instrument is operating normally (no alarm) the alarm pins connect as below.
Page 51: Switching On And Off
1. Plug the power cord into the mains socket (→ Fig. 27-1). 2. Make sure that the circuit breaker (→ Fig. 27-2) is at the position. 3. Press On the mains power switch (→ Fig. 27-3) in the front of the instrument. The instrument will take a few minutes to initialize. HistoCore PEGASUS...
Page 52: Switching Off
Setting up the Instrument 4. The login screen appears after the initialization completes. Input the user name and password. and the Status screen is displayed (→ p. 70 – 5.4 Status screen). 5. For the initial login (→ p. 58 – Initial login), use the Administrator account.
Page 53: Moving The Instrument
3. Release the castors of the instrument. 4. Support the corners of the instrument by hand and push it to a new location. 5. Lock the castors after the instrument has reached the final position. 6. Re-install the drip tray, the regent bottles, and the scanner (if the instrument is equipped with a scanner). HistoCore PEGASUS...
Page 54: Running Protocol
Running Protocol Running Protocol Using the software Instrument functions are controlled from the touch-screen. 5.1.1 Basic operation Buttons Tap buttons on the touch-screen to open menus, screens and dialogs, and to start and stop actions. The state of a button is indicated by its color or border. Depending on the screen and the operations available, enabled buttons (→ Fig. 28) could be dark gray, bright blue, bright pink, or bright green.
Page 55: Navigation
Some screens have alternative views to divide the information presented. For example, the Reagent Stations screen has two views, one for reagent bottles information and the other for paraffin chamber information. Tap buttons on these screens to move between the alternative views. HistoCore PEGASUS...
Page 56 Running Protocol In this Instructions for Use, navigation paths are shown in this way: > > This means from the function bar button, tap 1, then tap 2 and finally tap 3. For example, Administration > Protocols > means tap the Administration button on the function bar, tap Protocols (which will be on Administration...
Page 57: Access Levels
There are three user-access levels available to laboratory staff: Operator, Supervisor, and Administrator. Operators can carry out all routine tasks, such as running protocols and replacing reagents. However, operators cannot edit protocols or set reagent bottle configurations: you must have supervisor rights for these actions. Supervisors also have rights to configure the system. Administrators have all rights that Supervisors have and can create accounts for supervisors and operators. HistoCore PEGASUS...
Page 58 Running Protocol Some configuration settings displayed in the software require service-level access to change. These are provided so you can view the settings, but you must contact your authorized service representative if you want to change them. Initial login The Administrator account and its password is configured at the factory, which can be used to create new accounts. The default password is admin. 1. The login screen appears after the initialization completes. Tap Other access button (→ Fig. 36-1), and select Administrator.
Page 59: Help
• Only process patient tissue samples for diagnostic purposes, after all the above checks have been performed successfully. If any of the previous steps failed, do not operate the instrument and contact your local Leica Biosystems Service Representative. Warning User-defined protocols not validated.
Page 60 Running Protocol Warning Open retort while the instrument is processing/after pausing/after the protocol ends, the user gets contact with the hot surface of lid/baskets/fluids/hazardous reagents/vapor from the retort. Open the retort and the protocol cannot continue. People injury/Sample damage/Impaired samples • Hot surface. Wear chemical protective clothing, protective goggles, respirator, rubber gloves, and all other necessary personal protective equipment during operation.
Page 61 (for retort A) or bottom right (for retort B) to select a protocol from the Protocol selection screen. Only validated protocols (indicated by a green tick) (→ Fig. 38-1) can be selected by operator-level users. Fig. 38 HistoCore PEGASUS...
Page 62 Running Protocol B. If the first step of the protocol uses a reagent that is incompatible with retort residue, a warning message is displayed. You may need to run a cleaning protocol or you may be able to skip the first steps of the protocol (→ p. 80 – 5.5.2 Changing the starting step for a single run). If so, tap Clear all in the warning dialog box to continue. C.
Page 63 Fig. 40 The number displayed by default is set by supervisors. See (→ 8.3.2 Processing settings). However, it is important that you enter the correct number of cassettes. The figure you enter is used by the reagent management system to calculate reagent concentrations. Enter the number and tap OK. The scheduling screen will now appear. HistoCore PEGASUS...
Page 64 Running Protocol Fig. 41 11. If you want to edit any steps in the protocol, or change the duration of a step, tap Edit steps. The process is explained in (→ p. 80 – 5.5.2 Changing the starting step for a single run).
Page 65 Drain retort. 17. When prompted to, open the retort. 18. A message appears prompting you to remove all tissues. Do so and tap Done. Fig. 43 Do not tap Done until all baskets have been removed from the retort. HistoCore PEGASUS...
Page 66: Cleaning Protocols
Running Protocol Run cleaning protocol 19. The Clean now panel is now displayed. Run a cleaning protocol before running the next processing protocol. Warning Tissues are left in the retort when running the cleaning protocol. Sample damage • Remove all tissues from the retort before running a cleaning protocol as the dry step will damage the tissue.
Page 67 5. Continue with step 6 of creating a new cleaning protocol above. Cleaning protocols do not require a water step and work well with conventional cleaning reagents. To completely remove xylene from your instrument, Leica Biosystems can supply Waxsol™, a xylene-free cleaning solution (→...
Page 68 Running Protocol Warning Tissues are left in the retort when running the cleaning protocol. Sample damage • Remove all tissues from the retort before running a cleaning protocol as the dry step will damage the tissue. Warning Use cleaning protocol for sample reprocessing. Sample damage •...
Page 69 You will be prompted to do so. Do not put fresh cassettes into a retort that is showing residue in it, as indicated by the icon below (→ Fig. 44). (Formalin residue may be acceptable). Fig. 44 HistoCore PEGASUS...
Page 70: Status Screen
Running Protocol Warning Unprocessed samples (with formalin residue) are placed into a retort prior to running a cleaning protocol. Impaired samples - Formalin in the residue purged to the paraffin bath at the start of the cleaning run may damage tissue on subsequent runs. • Do not load unprocessed tissue samples into a retort prior to running a cleaning protocol. •...
Page 71: Status Area
— the retort is aborting the current action • Unavailable — the retort cannot be used. Contact your service representative. The retort has six possible states (→ Fig. 46-6). Icon Retort state Description Full The retort contains the correct amount of reagent or paraffin. HistoCore PEGASUS...
Page 72 Running Protocol Icon Retort state Description Part full The reagent or paraffin level is between full and empty. This usually occurs during a fill or drain operation. Empty The retort is drained but contains residue. Clean There is no residue in the retort. This only occurs after a cleaning protocol. The retort is now ready for another protocol run.
Page 73 4. Warning symbol and cross-hatching shows 2. Reagent type reagent is out of threshold 3. Color-coded by reagent group (see 5. Color or other pattern shows station state (→ p. 101 – 7.1.1 Reagent groups, types, (see below) stations)) 6. Reagent concentration HistoCore PEGASUS...
Page 74 Running Protocol Icon Bottle state Description Full The bottle holds sufficient reagent to fill a retort at the specified basket level. In use A reagent transfer is in progress or was aborted. The reagent level is between full and empty. Empty The bottle has been used to fill a retort. There is still reagent in the bottle. The bottle has been completely drained leaving only a small amount of residue.
Page 75: Protocol Panels
3. Select a protocol you want from the Protocol selection list. Tap the Select button on the right side of the screen. 4. The Select icon dialog box appears (→ Fig. 50). Select an icon for the protocol from the list, and tap OK. HistoCore PEGASUS...
Page 76 Running Protocol Fig. 50 5. The selected protocol shows in the Favorites list. Tap Save on the Favorites dialog box, and the selected protocol shows on the Status screen for both retorts. Move up (→ Fig. 49-2) Move down (→ Fig. 49-3) buttons to adjust the sequence of favorite protocols showing on the Status screen.
Page 77 9. Protocol finish time Unload protocol: unload protocols 10. Run/Pause: start a loaded protocol, or pause a (disabled while the protocol is running) running protocol In the expanded mode, the protocol notes, processing time, and step details are also shown. HistoCore PEGASUS...
Page 78: Protocol Run Options
Running Protocol Fig. 52 1. Step duration, temperature, P/V(Pressure/ 2. Processing time: total time to run protocol Vacuum), and stirrer ON/OFF Protocol run options You can schedule a protocol run so that it finishes at a convenient time. You can also modify the protocol to start at the second or later steps, and to change step durations. 5.5.1 Scheduling protocols Fig. 53 Version 1.6, Revision J...
Page 79 However, if there is a reagent clash caused by the protocol running in the other retort, the actual start of the protocol may be delayed (→ p. 86 – 5.7.1 Delayed end times and initial fills). • Set a new end time. Tap the Edit end time (→ Fig. 53-5) button to open the Enter required end time dialog box. (See the next section for instructions.) HistoCore PEGASUS...
Page 80: Changing The Starting Step For A Single Run
Running Protocol Entering a new end time or default scheduling setting Fig. 54 You can change the end time for the currently selected run in the Enter required end time dialog box (→ Fig. 54), opened from the Time to edit dialog box by tapping the Edit end time button (→ Fig. 53-5).
Page 81: Changing The Step Time For A Single Run
(→ Fig. 53) opens. Tap Edit steps (→ Fig. 53-6) to open the Edit the run steps dialog box (→ Fig. 55). 2. To change a step time, tap the current step time (→ Fig. 55-2). 3. Set a new duration in the dialog box. HistoCore PEGASUS...
Page 82 Running Protocol Fig. 56 4. Tap and repeat from step 2 above if you want to change any other step duration. Note • Be careful not to tap a step icon at the right of the dialog box. Tapping a step icon could change the protocol’s starting step.
Page 83: Pausing And Aborting Protocols
(→ Fig. 57) opens with a number of options: • Abort all processing or just the protocol in one retort. • Access a retort—for example to add more cassettes—and then resume the protocol running in it. • Vent the paraffin baths so you can open them. HistoCore PEGASUS...
Page 84 Running Protocol When the system starts to count down (→ Fig. 57-1), options are enabled for selection. You have five minutes to select an option, after which processing automatically resumes. Emergency stop and abort After you have paused the instrument you can abort all protocols with the Emergency stop button (→ Fig. 57-2).
Page 85: Recovering From An Aborted Run
(→ p. 86 – 5.7.1 Delayed end times and initial fills). When a second protocol starts, the reagent stations assigned when the protocol was loaded may change. This occurs because the first protocol’s reagent requirements always take precedence. HistoCore PEGASUS...
Page 86: Delayed End Times And Initial Fills
Running Protocol It is sometimes not possible to schedule a second protocol. This situation, and possible remedies, are discussed in (→ p. 86 – 5.7.3 Unavailable reagents). Also, protocols will sometimes fail if a reagent station unexpectedly becomes unavailable. See (→...
Page 87 However, if you choose to do so, you must make sure the bottle you intend to remove is not scheduled for use in either retort. • A paraffin station is not molten at the time it is required. Make sure there is adequate time for the paraffin to melt and that the correct paraffin station state is set (→ p. 116 – 7.3.2 Setting reagent station properties). HistoCore PEGASUS...
Page 88: Protocol Setup
Protocol Setup Protocol Setup Protocol overview A protocol consists of series of steps that will be applied to tissue in a retort. In each step (with one exception) the retort is filled with reagent and held for a time under conditions controlled by the protocol. Each protocol step specifies: • the reagent used • the time the reagent is in the retort •...
Page 89: Reagent Selection Method
Station selection rules The system uses the following rules to select stations for protocols with type-reagent selection. “Sequence”, as used below, means a series of protocol steps using the same reagent type. HistoCore PEGASUS...
Page 90: Pre-Defined Protocols
Administration > Protocols. The pre-defined protocols are designed for use with the bottle configurations (→ p. 162 – 10.3 Station configurations). Leica Biosystems has extensively tested the protocols and found they give excellent results (in combination with proper reagent management and instrument maintenance). However, users should validate all protocols, including the pre-defined protocols, for use in their laboratories, as different conditions could give different outcomes.
Page 91: Protocol Validation
The carryover due to cassettes and tissue is calculated using the number of cassettes entered by users at the start of each run. The amount of carryover on a standard cassette is used in the calculation. Drip time is also included. HistoCore PEGASUS...
Page 92 The following table shows the carryover values that should be used for a range of cassettes and other tissue carriers provided by Leica Biosystems. The values in the table apply when all the tissue in the run uses the respective cassette or carrier type.
Page 93 Protocol Setup Leica Biosystems products Generic description of tissue Carryover value if 100% of carrier tissue uses carrier ActivFlo Mini Cassette in Mini cassette in standard Surgipath ActivFlo Routine I cassette Surgipath Biopsy Wraps in Biopsy wrap in standard cassette...
Page 94: Protocol Files
Protocol Setup 6.1.6 Protocol files Protocols can be exported via USB in a database file with a .db extension. You can backup this file. You can also send the protocols to your service reprentative for troubleshooting. Backed-up protocol file can be imported into the HistoCore PEGASUS. See File transfer (→ p. 137 – Export files) for instructions on how to export and import protocol files. Creating, editing, and viewing protocols Supervisors can create and edit protocols. This is done from the Protocol selection screen. Operators cannot create or edit protocols but can view protocol details from the Protocol selection screen.
Page 95 7. Delete: remove the currently selected step 4. Save: save changes made to the protocol from the protocol 5. Notes: enter a description or other notes for 8. Done: confirm changes made to the protocol the protocol and return to the Protocol selection screen 9. Go to the previous/next row HistoCore PEGASUS...
Page 96 Protocol Setup Steps can be added and removed, and step details (such as the Step time, temperature, etc.) can be changed. The protocol’s name, notes, carryover setting and validation status can also be changed. However, you cannot change the protocol type or reagent selection method, nor can you remove the dry step from a cleaning protocol.
Page 97: Creating New Protocols
Only supervisors can create new protocols. You can create new protocols from scratch, or copy and modify an existing protocol (pre-defined or otherwise). Make sure you select the right protocol type and reagent selection method when you start (or copy a protocol of the right type and reagent selection method) because these settings cannot be changed after you have started to configure the new protocol. HistoCore PEGASUS...
Page 98 Protocol Setup Warning User defined protocols are not validated. Sample damage • The pre-defined protocols and the customized protocols have to be validated by the user before running the protocol, i.e. tissue processing with patient tissue for diagnostics, according to the local or regional accreditation requirements. Creating new protocols from scratch 1. Open the Protocol selection screen (Administration > Protocols) and tap New. 2.
Page 99 Validate protocol button. The protocol then becomes available to operators (provided that any other errors listed on the window have been attended to). Your new protocol will now appear in the protocol selection list. HistoCore PEGASUS...
Page 100: Viewing Protocols
Protocol Setup 6.2.4 Viewing protocols Operator-level users cannot create or edit protocols. However, they can view the details of a protocol (including step details, notes, and the date and time the protocol was last modified). 1. Tap Administration > Protocols. 2. Tap once on the protocol you want to view. 3. Tap View. Version 1.6, Revision J...
Page 101: Reagent Setup
Cleaning solvent First cleaning reagent Light pink Cleaning ethanol Second cleaning reagent Light blue A reagent is considered compatible in a protocol run based on its group (→ p. 109 – 7.1.6 Reagent compatibility). Types Reagent types are the specific reagents within each group, for example, formalin, xylene, Waxsol. As well as the chemical constituent, reagent-type definitions can include concentrations. For example, “70% Ethanol” and “80% Ethanol” are reagent types (and defined in this system). HistoCore PEGASUS...
Page 102 Reagent Setup Reagent types have the following properties: • Unique name • Default concentration: the concentration of the reagent when fresh. • Purity thresholds: to ensure that you replace degraded reagent (→ p. 104 – 7.1.3 Thresholds). • Temperature thresholds: used to ensure processing quality and safe usage of the reagent (→...
Page 103: Concentration Management
You should also update the software accurately whenever you change reagents. By default, the software assigns concentrations “by calculation”. This method uses the number of cassettes processed, the carryover setting, and the reagent groups involved to calculate the concentration in each station. HistoCore PEGASUS...
Page 104: Thresholds
Reagent Setup For consistently high-quality processing, always replace reagents as soon as you are prompted— with fresh reagent at the default concentration. If you remove a bottle from the instrument, always check that you enter the correct reagent information for the bottle when you return it. Supervisors can manually change station concentration values in the Reagent >...
Page 105 Final reagent thresholds set limits on the purity of the reagent that will immediately precede a change to another reagent group (or type, for protocols configured by type) in a protocol run. Final reagent thresholds are set higher than change thresholds. This ensures minimum contamination from the preceding reagent group to the subsequent reagent group. HistoCore PEGASUS...
Page 106 Reagent Setup When a reagent type or group has exceeded its final reagent threshold (that is, no bottle within that type or group is at or above the final reagent threshold) a warning symbol in an orange diamond appears on the least pure bottle in that type or group. In addition, a message appears above that bottle. After closing the message, you can redisplay it by tapping the warning symbol on the icon of the bottle. When you start the next run, a message appears informing you that this is the last run possible without changing the reagent that is below its final threshold: You can tap...
Page 107: Recommended Reagents
Compliance with the following guidelines will ensure superior tissue processing. The use of alternative reagents must be validated by your laboratory according to local or regional accreditation requirements. Adequate fixation of tissue samples is required for optimum results. This should be achieved prior to placing the samples in the system or incorporated into the fixative steps of the protocol. HistoCore PEGASUS...
Page 108 Reagent Setup The following reagents have been validated for use with the system. Fixatives • 10% Formalin (buffered and unbuffered) • Formal ethanol (70% ethanol and 30% formalin) Dehydration • 100 % ethanol • Reagent grade ethanol recommended for Histological Use •...
Page 109: Non-Recommended Reagents
(→ p. 104 – 7.1.3 Thresholds). They can also move reagents between the active list and dormant list, and create new reagent types. Operators can only view the active list. HistoCore PEGASUS...
Page 110: Pre-Defined Reagents
Reagent Setup 7.2.1 Pre-defined reagents A number of reagent types are pre-defined in the system. You can edit the properties of these reagent types, but they cannot be deleted. There is no requirement to use the pre-defined types; however, you can leave them on the dormant list. If you change the properties of pre-defined reagent type, you cannot use the software to automatically return it to its original values. 7.2.2 Editing active reagent types Use the Reagent types screen (Reagents > Reagent types) to view and edit the active reagent types list. You also access other reagent type management options from this screen.
Page 111 Reagent Setup Fig. 67 1. Selected reagent type 3. Defaults and concentrations 2. List of active reagent types 4. Reagent final thresholds Fig. 68 1. Selected reagent type 3. Defaults and concentrations 2. List of active reagent types 4. Reagent temperature thresholds HistoCore PEGASUS...
Page 112: Adding, Hiding, And Deleting Reagents
Leica Biosystems does not recommend that you lower reagent concentration thresholds. Raising concentration thresholds can help to correct poor quality processing due to impure reagents.
Page 113 The newly created reagent type is added to the list of dormant reagents. D. If you want to make a new reagent active, repeat from step 2 above; otherwise, tap Cancel. You are returned to the Reagent types screen. HistoCore PEGASUS...
Page 114: Managing Reagent Stations
Reagent Setup To hide or delete a reagent You can hide an active reagent by moving it to the dormant list. You can also delete a reagent. A deleted reagent no longer appears on either the active or the dormant list. (Only user-defined reagents can be deleted.) 1. To hide a reagent: A. Select it on the Reagent types screen.
Page 115: Reagent Stations Screen
6. Conc.: current concentration of the reagent in the station. Tap cell to change. Since changed: the use history of each station – view only Number: Reagent Lot number details. 9. State: the current state of the station. Tap cell to change. HistoCore PEGASUS...
Page 116: Setting Reagent Station Properties
Reagent Setup Fig. 71 Paraffin chambers button 2. Type: reagent type in the station. Tap cell to change. 3. Station: station number with reagent group color code. Hatched cell indicates change threshold exceeded. Out-of-threshold concentration value shown in red. 4. Selected station: tap in Station cell to select 5.
Page 117 If you select No you will retain the previous reagent’s concentration and history. Use this option only if you are correcting a previous error in the identification of the reagent in the station and are not actually changing the station’s contents. Warning Wrong definition of reagents Sample damage caused by reagents mismatch between GUI definitions and bottle labels. • Always update station details correctly. • Always check the definition of stations at/before the protocols start. • Never update the details without replacing the reagent. HistoCore PEGASUS...
Page 118 Reagent Setup Changing reagent concentration A supervisor can set a station’s calculated concentration value. Tap the station’s Conc. cell. Enter the new concentration using the on-screen keyboard. Warning Incorrect setup of the concentrations Sample damage or reduction in tissue processing quality •...
Page 119: Replacing Reagents
7.4.1 Remote fill/drain screen Use the Remote fill/drain screen (Reagents > Remote fill/drain), to fill and drain reagent bottles without removing them from the instrument. You use the same screen to drain the paraffin chambers. In a single operation, you can drain or fill a single station, or drain or fill a group of compatible stations. You can also fill and drain retorts from this screen. This enables you to recover from partially completed drain or fill operations. The functions on the Remote fill/drain screen (→ Fig. 74) are available to all users: operators and supervisors. HistoCore PEGASUS...
Page 120: Remote Fill/Drain Connections
Reagent Setup Fig. 74 1. Fill to/Drain from bottles, Drain to waste, or Fill/Drain retort: start fill or drain when appropriate station, remote source, and retort are selected 2. Abort: stop the drain or fill 3. Waste paraffin: select with a paraffin chamber to drain the chamber 4. Remote: select with a retort and bottle to fill or drain the bottle 5. Retort A: scheduled fill/drain operations for retort A 6. Retort B: scheduled fill/drain operations for retort B 7. Paraffin System: scheduled drain operations for the paraffin chambers 7.4.2 Remote fill/drain connections Fig. 75 The remote fill/drain line (→ Fig. 75-1) and the paraffin drain outlet (→ Fig. 75-2) sit beside the active carbon filter on the reagent cabinet. A protective flap (→ Fig. 75-3) covers the outlets. The paraffin waste line is heated to ensure that the paraffin does not solidify during the drain. Version 1.6, Revision J...
Page 121: Replacing Reagent - Remote Fill And Drain
• Always wear chemical protective clothing, protective goggles, respirator, rubber gloves, and all other necessary personal protective equipment, when handling the reagents. Reagents used for tissue processing can be both toxic and/or flammable. • Always use the tube supplied with the instrument. • Never fill a reagent station that has contents already in it. You can drain and then refill reagent bottles without removing the bottles from the instrument. The process drains each bottle to a retort and then drains the retort to the remote fill/drain line. The reverse procedure is used for the fill. HistoCore PEGASUS...
Page 122 Reagent Setup If you routinely use remote drain and fill, do not forget to check if bottles need cleaning. This should be done once a week. Before beginning a remote fill or drain, ensure both retorts are available. When a protocol is running in the retort(s), the remote fill/drain cannot be used. It is a design feature, not a malfunction. • both retorts must not have a protocol loaded or running • both retorts must be clean or empty and • both retorts must be clean or the residue in retorts (if any) must be compatible with the reagent in the bottle(s).
Page 123 To drain the retort, deselect the Remote icon then tap the Drain retort button. • Use a soft dry cloth to clean the surface of the tube before and after remote filling/draining reagents. Remote fill and drain sequences The following reagent sequences are recommended when filling and draining multiple bottles: Order Drain sequence Fill sequence Fixatives Cleaning solvents Cleaning ethanol Clearers Dehydrants Defatting solvents Defatting ethanol Defatting ethanol HistoCore PEGASUS...
Page 124: Replacing Reagent - Manually
Reagent Setup Order Drain sequence Fill sequence Defatting solvents Cleaning ethanol Cleaning solvents Dehydrants Clearers Fixatives 7.4.4 Replacing reagent – manually To replace a bottle manually, remove it from the reagent cabinet and dispose of the old reagent (following your laboratory’s standard procedures). Clean the bottle if necessary, and then fill it with fresh reagent. Load it back into the reagent cabinet, making sure that it fits firmly into the connectors at the back of the cabinet.
Page 125: Replacing Paraffin
5. Tap to begin the drain. The instrument will now drain the chambers. You can monitor the progress in the status panel. Paraffin drains can take up to 7 minutes. When the chambers are completely drained, a dialog will appear and the state of each drained chamber will be Dry. Do not remove the paraffin drain tube until the dialog confirms a successful drain. 6. To stop paraffin solidifying in the drain tube, promptly remove the tube from the container holding the drained paraffin. HistoCore PEGASUS...
Page 126 Reagent Setup Note • If no paraffin drains it is likely that the paraffin tube is blocked. If you remove a blocked tube before the drain is aborted, hot paraffin will spurt from the front of the instrument. Abort the drain before removing the tube and melting the paraffin with hot water. • At any time during the drain, you can tap Abort to terminate all current and pending fill/drain operations. Fill 7. If you are filling a paraffin chamber with molten paraffin, place the paraffin bath vent plug into the air vent at the rear of the paraffin chambers. This will help prevent paraffin from entering the vent during filling. Fig. 77 8. Fill the paraffin chamber with molten or solid paraffin. 9. If you placed the paraffin bath vent plug in the air vent, remove the tool. 10. Close the paraffin bath lid. 11. Go to Reagents > Stations > Paraffin chambers. The row for the drained paraffin chamber shows the default concentration. It also shows the history values re-set to zero. The station state is Dry. The reagent type is unchanged from the paraffin that you drained unless you changed the type assigned to the station after the drain.
Page 127: Filling And Draining Retorts
• You cannot fill a retort with a reagent that has a temperature threshold below the set retort temperature. • When draining a retort, the reagent/paraffin must be returned to its original station. • When draining a retort, the station must have sufficient capacity for the retort contents, and shall not be full or in use. Note: To avoid fluid spills, ensure that there is adequate station capacity before overriding insufficient capacity errors. To fill or drain a retort: 1. Go to Reagents > Remote fill/drain. 2. Select the retort that is to be filled or drained. 3. Select the reagent station that is to fill the retort (or collect the retort’s contents). 4. Tap the Fill retort Drain retort button. 5. To stop the fill or drain at any stage, tap Abort. HistoCore PEGASUS...
Page 128: Ancillary Settings & Operations
Ancillary Settings & Operations Ancillary Settings & Operations Reagents menu You can manually control a number of the reagent-handling features of the system from the Manual operations screen. 8.1.1 Manual operations From the Manual operations screen (Reagents > Manual operations) (→ Fig. 78) you can manually control many aspects of the instrument.
Page 129: Administration Menu
Fill (or drain) the retorts from (or to) a reagent station using commands on the Manual operations screen. The rules, and procedures, for manually filling and draining retorts are the same as when you do this from Reagents > Remote fill/drain screen (→ p. 127 – 7.4.6 Filling and draining retorts). Administration menu You can run reports, view event logs, and change access-level settings from the Administration menu. HistoCore PEGASUS...
Page 130: User Management
Ancillary Settings & Operations 8.2.1 User management Use the User management screen (Administration > User management) to manage user accounts. L The Barcode No. box will be enabled if the instrument works with the HistoCore I-Scan (Order No. 14 0488 58056). Adding/Editing/Deleting a user account L After the initial login (→...
Page 131: Reports
• Reagent usage report Run details report This report provides details of a particular protocol run. When selected, it first displays a list of all protocol runs, those in the previous month or those in the last 30 days (→ Fig. 80). Tap a button at the right of the screen to choose the duration you want. HistoCore PEGASUS...
Page 132 Ancillary Settings & Operations Fig. 80 From the protocol listing, you can display the details of one or more particular runs by selecting them and tapping Generate. The Run Details report appears, showing details of all the runs you selected (→ Fig. 81). The details provided include the start and end time, user ID, retort, number of cassettes, basket IDs, protocol steps, the station and reagent (with lot number if entered) for each step and the duration of the step. Any significant event that occurred during a protocol run is recorded.
Page 133 You can export the user action details by tapping Export. If no USB memory stick is already inserted, you will be prompted to insert a USB memory stick. The CSV file can be saved to a computer and opened in Excel. Protocol usage report This report lists the number of processing runs, cassettes, and average number of cassettes per run (→ Fig. 83). You can group the data by day, week or month. HistoCore PEGASUS...
Page 134 Ancillary Settings & Operations Fig. 83 You can export the protocol usage details by tapping Export. If no USB memory stick is already inserted, you will be prompted to insert a USB memory stick. An Excel-compatible file is generated. Tap Done when the transfer is complete. Reagent usage report This report lists the total volume of a reagent used by tracking when it is changed (→ Fig. 84). You can display the data by day, week or month.
Page 135: Event Logs
• To create a user event, select Create user event. This opens the on-screen keyboard. Type in the information you want to record and tap Enter to add the message as an event. The event severity will be “Information”. HistoCore PEGASUS...
Page 136: Settings Menu
Ancillary Settings & Operations Event severity There are three event severity levels and each has a color code: Color code Severity level Description Green Information A normal event that requires a response (e.g. a protocol successfully completed) or an unusual event that has no detrimental effect (e.g. a user aborted protocol).
Page 137 Set up the network connection (→ p. 142 – Network), and get optimized service from Leica Biosystems. The software area displays the current software version numbers. The software version may be needed when reporting a fault.
Page 138: Processing Settings
Ancillary Settings & Operations 8.3.2 Processing settings Warning The RMS (Reagent Management System) is set to N/A or a wrong value. The reagent consumption status can not be monitored, which may impact the tissue processing quality and cause sample damage. •...
Page 139 The system warns you if you begin an action that requires you to open a retort when the temperature inside it exceeds this value. Paraffin bath settings Paraffin type can be set as Recommended Recommended. • Set the paraffin type as Recommended for recommended paraffins (→ p. 107 – 7.1.4 Recommended reagents). The melting time is less than 7 hours. • Set the paraffin type as Not Recommended for other paraffins. The melting time is less than 9 hours. HistoCore PEGASUS...
Page 140: Instrument Settings
Ancillary Settings & Operations Global settings Set the paraffin melting and paraffin standby temperatures in the Global settings section of the Processing settings screen. • The paraffin standby temperature is the temperature the instrument heats the paraffin to in the paraffin stations. The default temperature is 65 ºC. • The paraffin melting point is the temperature at which the instrument considers the paraffin molten. The default temperature is 56 ºC. 8.3.3 Instrument settings Use the Instrument settings screen (Settings >...
Page 141 • To change the sound associated with a message type, tap the message type button in the Sound settings table. Select a sound file in the Select a sound from the following list dialog box and tap OK. To hear a sound in the Select a sound from the following list, select it and tap Play. HistoCore PEGASUS...
Page 142: Tracking Options (Optional)
Ancillary Settings & Operations • To change sound volume tap Master volume. Network For the initial connection to the remote service, contact the authorized service representative. Supervisors can connect/disconnect the instrument to Leica remote service if the Leica Remote Service is enabled on the Service settings screen...
Page 143 (→ Fig. 90-2) (→ Fig. 90-4) after the basket barcode is scanned during adding baskets into retorts. Picture view (→ Fig. 90-5) can be adjusted using the rotate buttons, clockwise (→ Fig. 90-6) and counterclockwise (→ Fig. 90-7). Delete the picture (→ Fig. 90-8) and reshoot if needed HistoCore PEGASUS...
Page 144: Cleaning And Maintenance
Ensure that the instrument is left on at all times, even when not in operation. This keeps paraffin molten and allows preventative maintenance routines controlled by the software to be carried out. Leica Biosystems recommends a preventative maintenance call once a year. Contact your service representative to organize a call. Do not use any cleaning materials or methods other than the ones described in this chapter. The use of other materials and methods may damage the instrument.
Page 145: Maintenance Screen
To record a maintenance task: • tap the panel for the maintenance task you have done or are about to do • tap Done. Recorded maintenance tasks can be viewed in the User action report (Administration > Reports > User action). HistoCore PEGASUS...
Page 146: Cleaning And Maintenance Schedule
Cleaning and Maintenance Screen maintenance To lock the screen, tap the Lock for cleaning button. The clean screen dialog box opens. You can safely clean the screen without accidentally selecting any controls while this dialog box is open. When finished cleaning, tap the clean screen buttons in the correct order. This unlocks the screen and returns it to normal operation. Cleaning and maintenance schedule See the list below for regular cleaning and maintenance tasks and how often they should be done.
Page 147 Bottle 16 Bottle 17 Paraffin 1 Paraffin 2 Paraffin 3 Paraffin 4 Clean top surface Clean touch-screen Check drip tray Weekly Tasks Empty condensate bottle Check reagent bottles Clean paraffin bath Check bottle connectors Clean exterior surfaces Every 60–90 Days Change active carbon filter Check lid seals HistoCore PEGASUS...
Page 148: Daily Tasks
Cleaning and Maintenance 9.3.1 Daily tasks Clean seals (paraffin bath & retort) and air vents Use the plastic scraper provided to scrape paraffin off the inside surfaces of the retort and paraffin bath lids. Replace any seals that are damaged (→ p. 156 – Check lid seals). Scrape around the rims of the retorts, and around the paraffin bath on the top of the instrument where the lids sit. Ensure that the lids seal fully when closed. In the paraffin bath, make sure that the air vent (→ Fig. 92-1) on the back rim under the left lid is clear of paraffin. Use the paraffin bath vent plug to block the paraffin air vent while cleaning. This will prevent paraffin from falling into the paraffin vent.
Page 149 • If you cannot remove stains with ethanol solution, use a lint-free cloth moistened with 6% acetic acid or CLR®. Wipe the surfaces again with ethanol. • Inspect the filter (→ Fig. 95-3). If soiled, then a. Release the retort filter by rotating it upwards. b. Wash it thoroughly in a solution of 70% ethanol. c. Rotate it back into place. HistoCore PEGASUS...
Page 150 Cleaning and Maintenance Warning Blockage by losing cassettes/tissues from retort or impurities from reagents or paraffin Sample damage • Use baskets during processing. Do not put the specimens into the retort directly. • Check that the sieve is installed at the retort bottom before running a protocol. • Take care not to drop anything into the retort when the retort sieve is removed. Foreign material in the pipes could stop valves from operating properly.
Page 151 A pop-up screen with instructions is available by tapping Help. Check paraffin fill levels Ensure that the paraffin baths are filled to between MIN and MAX marks. Add more paraffin if required (→ p. 119 – 7.4 Replacing reagents). Do not fill above the MAX mark. When complete, record this maintenance on the Maintenance screen by tapping the appropriate panel: Fig. 98 A pop-up screen with instructions is available by tapping Help. HistoCore PEGASUS...
Page 152: Weekly Tasks
To clean the bottles, pour in a small amount of fresh reagent (i.e. the reagent the bottle is used for), cap the openings, and shake. Use sealing caps from Leica Biosystems to seal the bottle. Empty the bottle and check that it is clean. If so, refill it and replace it in the instrument. See (→ p. 119 – 7.4 Replacing reagents) for instructions to reset the station properties.
Page 153 Maintenance screen by tapping the appropriate panel: Fig. 101 A pop-up screen with instructions is available by tapping Help. To avoid splashes when cleaning bottles, wear suitable protective clothing and eye-wear. To avoid reagent spills, ensure that lids are tight and the bottles are firmly plugged into their connectors at the back of the reagent cabinet. HistoCore PEGASUS...
Page 154 Cleaning and Maintenance Do not clean reagent bottles in an automatic dishwasher, as they may be damaged. Check paraffin baths Check the paraffin baths weekly to see if they are becoming dirty. Clean the baths when you are next replacing the paraffin. Clean the paraffin baths while they are warm (as they will be immediately spent out after having been drained from the instrument). Never attempt to clean a bath while the paraffin from the bath is in a retort. Remove any debris or sludge from the bottom and sides of the bath by wiping them with a lint-free cloth. Be careful not to dislodge or remove the chimney filters. Use the paraffin bath vent plug to ensure that paraffin does not go down the paraffin bath vent.
Page 155: Days
Settings > Instrument settings. To replace an active carbon filter: 1. Open the remote fill/drain cover. 2. Remove the active carbon filter by pulling its woven belt on the front side. 3. Remove a new filter from its plastic wrap. 4. Insert the new filter into the instrument. The date on which the filter has been inserted can be marked on the label on the front side. 5. Close the remote fill/drain cover. When complete, record this maintenance on the Maintenance screen by tapping the appropriate panel: HistoCore PEGASUS...
Page 156 Cleaning and Maintenance Fig. 102 A pop-up screen with instructions is available by tapping Help. Check lid seals Every 60–90 days, inspect the seals around the retort lids and paraffin bath lids. Call service to replace a seal if it is worn or damaged. Version 1.6, Revision J...
Page 157: Reference
Formalin 98.0 85% ethanol 50.0 80/20 ethanol/IPA 81.0 90.0 95.0 Paraffin 85.0 95.0 Cleaning Waxsol 6 cycles* Cleaning ethanol 6 cycles* * This differs from the recommendation for cleaning ethanol in xylene processing because the cleaning solvent in xylene-free protocols is less efficient than the cleaning solvent in xylene protocols. HistoCore PEGASUS...
Page 158: Protocols
Reference 10.2 Protocols The protocols in this section have been developed, and extensively tested, by Leica Biosystems for use on the tissue processor. Some are included as pre-defined protocols with the system. When used for the recommended tissue types, the protocols all produce optimum processing quality with consistent, high-quality results. Use these protocols and the suggested station configurations (→ p. 162 – 10.3 Station configurations) as a reference point when developing protocols that suit your specific requirements and practices.
Page 159: Xylene Protocols
(min) Formalin Ambient Ambient Ethanol Ambient Ambient Ethanol Ambient Ambient Ethanol Ambient Ambient Ethanol Ambient Ambient Ethanol Ambient Ethanol Ambient Xylene Ambient Ambient Xylene Ambient Ambient Xylene Ambient Paraffin Vacuum Paraffin Vacuum Paraffin Vacuum Processing time: 3hr 11min HistoCore PEGASUS...
Page 160 Reference Overnight xylene • Pre-defined standard protocol “Overnight Xylene” • Carryover setting: 25 Step Reagent type Time (min) Temp (°C) Stirrer Drip time (s) Formalin Ambient Ethanol Ambient Ethanol Ambient Ethanol Ambient Ethanol Ambient Ethanol Ambient Ethanol Ambient Xylene Ambient Xylene Ambient Xylene Ambient Paraffin Vacuum Paraffin...
Page 161: Xylene-Free Protocols
Time (min) Temp (°C) Stirrer Drip time (s) Formalin Ambient Ambient 85% Ethanol Ambient Ambient 85% Ethanol Ambient 80/20Ethanol/IPA Ambient Ambient 80/20Ethanol/IPA Ambient Ambient Ambient Ambient Ambient Ambient Paraffin Vacuum Paraffin Vacuum Paraffin Vacuum Processing time: 2hr 59min HistoCore PEGASUS...
Page 162: Cleaning Protocol
Reference Overnight xylene-free • Pre-defined xylene-free protocol “Overnight Xylene-free” • Carryover setting: 26 Step Reagent type Time (min) Temp (°C) Stirrer Drip time (s) Formalin Ambient 85% Ethanol Ambient 85% Ethanol Ambient 80/20Ethanol/IPA Ambient 80/20Ethanol/IPA Ambient Ambient Ambient Ambient Paraffin Vacuum Paraffin Vacuum Paraffin Vacuum Processing time: 12hr 11min...
Page 163 Cleaning Ethanol Paraffin Paraffin Paraffin Paraffin *All ethanol dehydrants after fixative should be labeled and named in the software “Ethanol”. However, if setting up bottles on a new instrument or replacing all the ethanol bottles at once, the first and second bottles should be filled with 70% and 90% ethanol respectively. As operation continues and you are prompted to replace degraded bottles, replace with 100% ethanol. Other, initially higher-concentration bottles will have degraded, so there will always be at least one bottle at a low concentration. HistoCore PEGASUS...
Page 164 Reference Xylene-free configuration This is the suggested station configuration when running xylene-free protocols. See the note below the table for special conditions that apply to the initial bottle setup and when replacing all the 85% ethanol bottles at once. Station Reagent Formalin Formalin 85% Ethanol * 85% Ethanol * 85% Ethanol 85% Ethanol 80/20 Ethanol/IPA 80/20 Ethanol/IPA 80/20 Ethanol/IPA Cleaning xylene...
Page 165: Protocol Step Temperatures
Manual operation Current Step Previous Step Y = Compatible None Fixatives Processing Dehydrants Defat Post Clearers Paraffin Cleaning Cleaning Water Defat Solvents Ethanol Fixatives Processing Water Dehydrants Defat Post Defat Clearers Paraffin Cleaning Solvents Cleaning Ethanol HistoCore PEGASUS...
Page 166 Reference Standard processing Current Step Previous Step Y = Compatible None Fixatives Processing Water Dehydrants Defat Post Defat Clearers Paraffin Fixatives Processing Water Dehydrants Defat Post Defat Clearers Paraffin Cleaning protocol Current Step Previous Step Y = Compatible None Fixatives Processing Dehydrants Defat...
Page 167 IPA (Group) Paraffin *Reagent type will be selected during creating processing protocol in xylene-free mode. Xylene-free reverse processing (purpose to remove paraffin) Current Step Previous Step Y= Compatible Cleaning Solvents Cleaning Ethanol None Dehydrants IPA (Group) Paraffin Cleaning Solvents Cleaning Ethanol Dehydrants IPA (Group) Paraffin HistoCore PEGASUS...
Page 168: Troubleshooting
Troubleshooting 11. Troubleshooting 11.1 Preliminary questions When faced with blocks that are difficult to section, first collect as much information as possible to assist in analyzing the problem. Some questions to ask are: 1. Did the instrument software indicate an error in the run? Instrument warnings and alarms will be displayed on screen and in the event log. 2.
Page 169: Under-Processed Tissue - Instrument Setup
Fixation time Is the tissue workflow • Water introduced after processing? processed appropriate? • Quality of embedding wax correctly? Are these all correct? Stop: Go to 11.2.3 Under- or Over- well processed tissue processed Tissue – Reagents Fig. 103 HistoCore PEGASUS...
Page 170: Over-Processed Tissue - Instrument Setup
Troubleshooting 11.2.2 Over-processed tissue – instrument setup Over-processed tissue (cooked/ crispy/ burnt) If a user warning, fix the issue. Retest. Is the Are any error If a service warning, contact tissue processed codes present? service correctly? Check: • How was the tissue submitted to the lab (in formalin)? •...
Page 171: Under- Or Over-Processed Tissue - Reagents
• failing to clean the retorts before or after remote drain or fill operations Open a service call to have Leica Biosystems Is the wax test the instrument and repair if required. contaminated or does it smell like...
Page 172: Poor Processing - Incorrect Protocol
Troubleshooting 11.2.4 Poor processing – Incorrect protocol Poor processing – incorrect protocol Try: Tissue is over- Was the • soaking the block in a softening agent, such processed. original protocol as fabric softener, before cutting. Reprocessing will not too long? •...
Page 173: Poor Processing - Correct Protocol
Try a softening agent, such as fabric softener, at sectioning. If it is a foreign body such as a staple, melt block down, remove object and re-embed Fig. 107 HistoCore PEGASUS...
Page 174: Cutting Artifact
Troubleshooting 11.2.6 Cutting artifact Cause Suggestion for fix Over-dehydration Decrease time in dehydrant Is the tissue hard, brittle or Over-clearing Decrease time in xylene has chatter? Warm cold plate slightly or wet face Cold plate too cold of block just prior to cutting Check knife tilt and ensure proper Poor microtomy cutting techniques...
Page 175: Staining Artifact
Troubleshooting 11.2.7 Staining artifact Fig. 109 HistoCore PEGASUS...
Page 176: Block Artifact
Troubleshooting 11.2.8 Block artifact Fig. 110 Version 1.6, Revision J...
Page 177: Reprocessing Recommendations
• Isotonic saline • Sandison’s solution a. 50 mL 1% aqueous formalin b. 30 mL 96% ethanol c. 20 mL 5% sodium carbonate • Ethanol–glycerol a. 90 mL 60% ethanol b. 10 mL glycerol Then process the tissue normally. HistoCore PEGASUS...
Page 178 Troubleshooting 4. Hard shriveled tissue – dried out but processed through to paraffin Remove paraffin and rehydrate as detailed in problem 6, below. Apply a reconstitution solution as described in problem 3. 5. Tissue insufficiently infiltrated with paraffin Place the cassettes back into the paraffin bath with vacuum, stirrer on and temperature at 65 ° C. This may be necessary when tissue has been prematurely removed from the paraffin. 6. Adequately fixed Under-processed tissue This problem can be due to too short a protocol, too large a specimen, or processor failure. Four solutions are recommended, but first identify the problem and rectify it. Test your fix by running control tissue through the processor before reprocessing the patient tissue (or use another processor). Note: For the following solutions, first melt the blocks, blot off excess paraffin, and then place the specimens in new cassettes. This minimizes paraffin contaminating the processing reagents.
Page 179 (→ p. 158 – 10.2.1 Specimen type and protocol duration). Unfixed or poorly-fixed tissue is damaged by processing. The ethanol and high temperatures exert a fixative effect different to that of formaldehyde. As a result, blocks become more susceptible to hardening and shrinkage. When combined with under-processing, blocks may be unusable. Rapid reverse cleaning protocol Step Reagent type Time (min) Temp (°C) Stirrer Drip time (s) Cleaning solvents Ambient Cleaning ethanol Ambient Processing time: HistoCore PEGASUS...
Page 180: Power Failure
Troubleshooting Slow reverse cleaning protocol Step Reagent type Time (min) Temp (°C) Stirrer Drip time (s) Cleaning solvents Ambient Cleaning solvents Ambient Cleaning ethanol Ambient Cleaning ethanol Ambient Processing time: 11.3 Power failure If there is evidence of a power failure to the instrument, check for error messages indicating the nature of the problem after the power is restored.
Page 181: Ordering Information
14 3000 00065 M6x35 ISO4762 fully thread 14 3000 00537 Straight Screwdriver 14 0488 60138 Ratchet spanner 17mm 14 0488 60142 Exhaust tube 14 0422 31975 Note • The above accessories may not be available in some countries/regions. HistoCore PEGASUS...
Page 182: A1 Decontamination Confirmation
Decontamination Confirmation A1. Decontamination Confirmation Any product that is to be returned to Leica Biosystems or serviced on site must be properly cleaned and decontaminated. The associated decontamination certificate template can be found on our website www. LeicaBiosystems.com in the product menu. This template must be used to enter all required data. If a product is returned, a copy of the completed and signed decontamination certificate must either be enclosed or handed over to a service technician. The user shall be responsible for products that are returned without a completed decontamination certificate or with a missing decontamination certificate.
Page 183: A2. Warranty And Service
Warranty and Service A2. Warranty and Service Warranty Leica Biosystems Nussloch GmbH guarantees that the contractual product delivered has been subjected to a comprehensive quality control procedure based on the Leica in-house testing standards, and that the product is faultless and complies with all warranted technical specifications and/or agreed-upon characteristics. The scope of the warranty is based on the contents of the contract concluded. The warranty terms of your Leica sales organization or the organization from which you have purchased the contractual product shall apply exclusively.
Page 184 Notes Version 1.6, Revision J...
Page 186 Leica Biosystems Nussloch GmbH Heidelberger Strasse 17 - 19 69226 Nussloch Germany Tel.: +49 - (0) 6224 - 143 0 Fax: +49 - (0) 6224 - 143 268 Web: www.LeicaBiosystems.com...

Leica BIOSYSTEMS HistoCore PEGASUS Instructions For Use Manual

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