Dako Autostainer | English Handbook
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Dako Autostainer Handbook Document Number 0000620 Revision A July 2006 Autostainer Handbook iii
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Copyright © 2006 Dako, Inc. All rights reserved. This document may not be copied in whole or in part or reproduced in any other media without the express written permission of Dako, Inc. Please note that under copyright law, copying includes translation into another language. User Resources For the latest information on Dako products and services, please visit the Dako Web site at: http://www.dako.
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Table of Contents Warnings, Precautions and Limitations............................................................................. 1 System Overview .............................................................................................................. 5 Software Overview ............................................................................................................ 7 Sign In Screen.........................................................................................................
Negative Controls.................................................................................................... 21 Defining Specific Rinses ............................................................................................. 22 Auto Programming ...................................................................................................... 22 Deleting an Auto Programming Item ....................................................................... 23 Using an Auto Programming Item ......
Section 1 Warnings, Precautions and Limitations Keep the cover closed during operation. The robotic arm will move unexpectedly during the operation - stay clear. Do not impair the movement of the Autostainer robotic arm in any way. Do not pour liquid down the Autostainer sink. The system is not equipped to drain large liquid volumes poured at high speed. Hazardous reagent wastes must be disposed of according to local, state, and federal regulations.
Do not print slide labels while the Autostainer is running. Immunohistochemistry is a multi-step diagnostic process that requires specialized training in the selection of the appropriate reagents, tissue selection, fixation, and processing, preparation of the IHC slide, and interpretation of the staining results. Tissue staining is dependent on the proper handling and processing of tissues prior to staining.
Correct Disposal of this Product (according to Directive 2002/96/EC on Waste Electrical and Electronic Equipment [WEEE] applicable in the European Union and other European countries with separate collection systems). Contact a Dako representative for disposal of the equipment at the end of its working life. This product should not be mixed with other commercial waste for disposal.
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Section 2 System Overview The Autostainer System is an automated slide processing system compatible with currently available reagents for staining paraffin-embedded and frozen tissue sections, cytospins, cell smears, and fine-needle aspirates. This system is designed to automate manual staining methods routinely used in immunohistochemistry and cytochemistry, enabling the transfer of established protocols from the bench to the Autostainer.
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Section 3 Software Overview Sign In Screen The Sign In screen appears when you double-click the Autostainer icon on the desktop. After your name and password have been validated, the Main Menu appears. Main Menu The buttons on the Main Menu allow you to access the other Autostainer screens. Note: Change Password replaces the Initialize button for users with middle and low security access.
Initialize The Initialize screen is used to establish and update default information for the system. This includes the institution information, Autostainer serial number, user names, the doctors requesting IHC tests, a library of tissues that may be used during staining, the default volume and the drop zone for reagent dispensation, and the number of slides allowed between routine cleaning runs.
2. Select the desired option for each field. Note: Static information, such as the institution name, can be entered into the field. This information is printed on every label. To enter static information, highlight the text in the desired field and type the text in the text box. Press ENTER. 3. Select the number of characters for each line (10, 12, 14, or 16) by selecting a number from the Length list (Selecting 10 characters will display bold text).
Printer Selection To select the proper Seymour printer type on the Design Slide Label screen, scroll though the printer list using the keyboard arrows or the scroll bar. Note: If the incorrect printer is selected, error messages display on the screen when slide or reagent labels are printed. To find the model number of your Seymour Glass Label printer, check the cover of the printer for a number (e.g.,TPL2642). The last four digits represent the model number (e.g., 2642).
Add Rinse to Protocol Steps: When creating a new protocol template, a buffer rinse can be added automatically, following each step. Select the Add Rinse to Protocol Steps option to automatically add buffer rinses. If this option is not checked, the rinse step must be manually selected. No Pre-Rinse: When creating a new protocol template, the default pre-rinse step may be removed. If this option is not checked, the protocol template will include a buffer rinse step.
Prime Pump The Prime Pump screen is used to flush the intake tubing with buffer or DI water without initiating an actual run. Frequently running fluid through the instrument keeps the Autostainer running properly. Dako recommends priming the pumps weekly. Programming Grid Overview The Programming Grid is the main screen used to define slide information, primary antibodies, staining reagents, and staining protocols.
Section 4 Entering Slide Information Slides can be programmed from the Programming Grid either through Slides on the menu bar for quick programming of total slide count, or through the Slide Info button for entering specific slide identification information. Adding Slides The Slides option expedites programming by reducing the amount of information required to program a staining run. 1. Select Slides on the menu bar.
Deleting Slides Deleting Slides With the Slides Function When using the Slides option on the menu bar to program a staining run, a slide can be deleted if there is at least one entirely blank (unprogrammed) row. Select the Slides menu item, adjust the number of slides, and click OK. Note: If all rows are programmed, the slide must be deleted from the Programming Grid (see Deleting a Specific Slide, later in this section).
Section 5 Designing a Protocol Protocol templates allow you to enter the series of steps required to stain a slide and save the protocol for future use. Low level security does not allow creating or editing protocol templates. Standard Protocol Elements Protocol elements are categories of reagents used to build the protocol outline in the Protocol Template Design screen. Reagents can be added, deleted and edited by choosing protocol element categories from the Edit Lists menu item on the Programming Grid.
Additional Protocol Elements Rinse Buffer A buffer rinse step consists of a buffer wash and air blow cycle and should be programmed between each protocol template step. Each buffer rinse is displayed as a blue droplet in the programming grid with a droplet icon in each programmed row. The buffer icon appears on the Programming Grid printout as a filled-in droplet. Rinse Water A water rinse step consists of a water wash and air blow cycle.
Note: If No Pre-rinse is selected at the Options screen, the first rinse buffer is not displayed (See Initializing, Section 3). 3. Build the new protocol template from the Protocol Elements list. Click on a reagent in the list, the item is copied to the Protocol Outline list and sequentially numbered. Note: If Add Rinse to Protocol Steps is selected at the Options screen, rinses are automatically added after each step, except following a substrate step. 4.
3. Click the Use Template button. The Programming Grid appears with the selected protocol template displayed in the grid header. Using a Saved Protocol Template for a Specific Auto Program Auto Programming is a feature for making slide programming more efficient (see Programming a Staining Run, Section 6). 1. 2. 3. 4. 5. Select the Protocol Template button in the Programming Grid. Select the Get Template button. Select the Auto File check box.
Section 6 Programming a Staining Run The Autostainer can process a maximum of 48 microscope slides in a single staining run. Each slide can be stained using an independently designed protocol. After the slide information is entered and protocol template is selected, specific reagents are assigned to each slide. This is accomplished from the Programming Grid. The following section describes how to program specific reagents for a staining run. Each row represents one slide and its information.
4. Select Yes and, if the program is new, a dialog box prompts “Save program on disk?” Otherwise, the Slide Layout Map screen appears. Select Back to Grid to return to the Programming Grid and the missing rinse appears in the grid. Defining Primary Antibodies Each primary antibody reagent is programmed individually and will appear in the Programming Grid with its programmed pretreatment (see Reagent Management, Section 7).
Adding Positive and Negative Control Reagents 1. Enter Slide Information and select a protocol template. A minimum of one slide must be programmed through Slide Count or the Slide Information screen. Click OK or the Finish Entry button. 2. Fill in Programming Grid columns with specific reagents for one slide. After the primary antibody is applied, Neg.Ctl and Pos.Ctl buttons display in the menu bar. Continue entering the detection reagents. 3. To insert a matching positive control, click the Pos.Ctl button.
antibody does not have either a compatibility or negative control type defined, an error appears stating that there is no matching negative control reagent. Note: If you do not wish to use this feature noted above, then continue programming as usual. This feature is not active when using Auto Program. Defining Specific Rinses A rinse is automatically added following every programmed reagent (except for Protein Block, where a blow step is added) when assigned as described above.
Deleting an Auto Programming Item 1. Select Auto from the menu bar on the Programming Grid. 2. Select Setup from the Auto menu. The word Auto changes to Setup. 3. Select any activated tile in the Programming Grid to display the Save Auto Program window. 4. Click the Auto Program to be deleted. 5. Click the Delete Program button. 6. Click the Yes button to delete the program. Using an Auto Programming Item 1. Select Auto from the menu bar in the Programming Grid. 2. Select Program from the Auto menu. 3.
Editing a Protocol Step The Autostainer software provides multiple methods for editing the reagents in a defined staining run from the Programming Grid. Changing a reagent for a step: 1. Click the reagent step you wish to change. 2. Select the desired reagent from the list. The new reagent is displayed for the step. Replacing a reagent with skip step (no reagent added to slide): 1. Click the reagent step you wish to change. 2. Select the word None. Editing a reagent in a staining step for one run only: 1.
Reagent Labels Printing reagent labels from a programmed run: 1. Program or retrieve a previously saved run on the Programming Grid, and select the Print button. 2. The Which Report screen appears, click the Reagent Labels button. 3. To print all reagent labels, select the Print All button. 4. To print specific reagents, click the desired reagents and select the OK button. Printing reagent labels not associated with a specific program: 1. 2. 3. 4. 5. 6. From the Programming Grid select the Print button.
Assigning Reagent Dispense Locations The non-frosted area of each slide is equally divided into three horizontal sections – or dispense locations. Each slide can have one to three dispense locations, and the maximum dispense volume of a slide cannot exceed 800 µL. The default dispense location is programmed during initialization.
Section 7 Reagent Management Reagent lists are created and modified using the commands on the Edit Lists menu on the Programming Grid. Adding Detection Reagents (Secondary, Tertiary, Substrate, etc.) 1. From the Programming Grid select Edit Lists from the menu bar. 2. Select the reagent category from the list. 3. Enter the full name of the reagent you want to add to the current list and press ENTER. The software allows the entry of 33 alpha-numeric characters for detection reagents.
Adding Primary Antibodies A primary antibody can be added to the reagent list with a related pretreatment reagent. This facilitates automatic selection of the pretreatment whenever a primary antibody is selected from the Programming Grid. 1. Follow the same steps as in the Adding Detection Reagents section. Note: To add a matching negative control use exactly the same name as the positive control and add the prefix “_NC” to the beginning of the reagent name. 2.
Compatibility Check A compatibility check is included to minimize programming errors resulting from assigning incompatible reagents (such as alkaline phosphatase-labeled streptavidin and DAB substrate) to a slide protocol. This feature has two check levels. The first check is for the species reactivity compatibility of the primary antibody and the secondary reagent. The second addresses the compatibility rules dictated by the enzyme used in the detection system.
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Section 8 Loading Reagents After programming is complete, the Autostainer calculates the time required for the staining run, the amount of wash buffer or deionized water required, and the most efficient placement for the reagent vials. Reagent Layout Map Screen The Reagent Layout Map documents the rack positions of the reagents including the fill volume needed for completing the staining run. It displays a maximum of 32 reagent vials per rack.
5. Check that the reagent vials have no bubbles on top of the solution. Remove any bubbles. Missing Reagent Notice The Autostainer displays a prompt when an insufficient volume of reagent is detected. You can pause the system to add more reagent to the specified vial. If sufficient volume of a reagent is not available, and you do not respond to the alarm, the Autostainer continues the run and provides a list of skipped or partially applied slides on the Run Log.
Section 9 Loading Slides After programming is complete and the reagents are loaded, you are ready to load the slides on the slide racks. Print the Slide Layout Map: Load Slides screen by selecting the Print button at the bottom of the screen. This map provides a guide for loading slides and can be saved as part of your staining record. Note: Soak slides in a buffer solution containing 0.05% Tween-20 for a minimum of five minutes prior to loading the Autostainer.
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Section 10 Starting a Staining Run Preparing the System for a Staining Run After the specimen-containing slides and reagent vials are loaded, perform the following checklist: Slides • Deparaffinized and rehydrated, soaking in buffer containing 0.05% Tween 20 using a squirt bottle to prevent slides from drying while loading additional slides. • Seated securely in slide rack. Racks • Properly seated in the Autostainer. Reagents • At room temperature. • Placed at proper locations in the reagent racks.
6. Select the Protocol Template button and select a protocol template for the run. 7. Select the Use Template button. The Programming Grid appears with the selected protocol. 8. Select the desired volume and drop zones for either all slides, or on an individual side basis. 9. Assign specific antibodies and reagents to all slides and select the Next button. The Slide Layout Map: Program Slides screen appears. 10. Select the Next button and the Reagent Layout Map appears. 11.
Section 11 Maintenance and Troubleshooting The Autostainer should be cleaned after 150 slides have been processed or after one week, whichever comes first. The instrument keeps track of the number of staining runs performed and displays a maintenance message listing the number of slides since the last cleaning cycle. Note: Please contact your Dako representative regarding maintenance details. Troubleshooting Problem Autostainer will not turn on (top green LED not on).
Problem Buffer or DI Water does not flow out during a run or a prime. (Cont.) Possible Cause Tubing is disconnected at the inlet fitting(s). Inlet filters are clogged and are restricting fluid flow. Tubing not completely inserted into carboy. All waste collects into one waste container. Faulty buffer / DI water pump. “Switch” is not programmed in the Protocol Template. Waste tubing not placed in the proper waste carboy. Faulty waste pump. Waste backs up in the sink. Clogged sink filter.
Problem The probe and wash/blow head are not going to the correct locations. No Staining Possible Cause The arm has been knocked out of calibration. Course of Action Abort run and save run log. Reprogram the run and delete the steps from the Protocol Design screen that have already been completed. Restart run. Bent probe tip. Verify that all reagent vial caps have been removed and slide racks are firmly locked in place in the down position. Contact a Dako representative.
Problem Slide number is not listed on label. Computer functions lock up during a run, but the Autostainer is still performing the run. User is unable to access many of the functions in the Initialize section of the program. Illegal or fatal exception errors. Possible Cause Label is not centered correctly on label. Windows error. Course of Action Reprint label if necessary and place the label 1/2 cm from the top and 1/8 cm from the side of the slide. User has been set up as a “NonAuthorized” user.
Problem Words on your monitor screen are too dim. Image on the desktop is grainy and the color is incorrect. Possible Cause The brightness control is not set properly. Windows started in “Safe Mode.” Color depth has been adjusted too low. The printer does not respond to print command. The printer may be in Manual Feed mode. The parallel cable between the printer and the computer is not connected properly. The parallel cable is defective.
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Section 12 System Specifications Hardware Specifications Dako Autostainer Dimensions 1.02m W x 0.69m D x 0.61m H 40" W x 27" D x 24" H Weight 140 lb. (63.
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Section 13 Standard Configuration Ensure that the area for the instrument is a solid, level surface that can support the weight of the system (approximately 240 lbs.) Minimum work area dimensions: 2.0 m W x 0.8 m D x 1.0 m. H The work area must have an ambient temperature between 18°C – 26°C (64°F – 79°F), not facing direct sunlight and not effected by a hood or vent. Dako recommends a dedicated power source to prevent interference from other instruments or equipment.
