1. 云顶集团3118(中国)

    U54 SCENT Histology H&E Staining

    Shannon McCall

    Published: 2023-06-12 DOI: 10.17504/protocols.io.4r3l22m9ql1y/v1
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    Abstract

    To provide a standardized method of running, cleaning, refilling, and verifying quality of the automatic H&E stainer. Every morning before running slides for projects, a quality control slide containing brain, kidney, liver, and pancreas tissue is run, reviewed, and deemed acceptable by a histotechnician, who documents this review.

    Steps

    Protocol - Operating Autostainer and Running Daily Quality Control

    1.

    Open cover of autostainer and remove each lid of the reagents inside. Stack lids and set to the

    right side of autostainer. Close cover.

    2.

    Hit switch on the right side of autostainer to turn on and initialize.

    3.

    Once initialization is complete, press button on screen to select staining program 1.

    4.

    Take one slide from the box of unstained H&E control slides and label with:

    a. H&E QA
    
    b. Date
    
    c. Tech initials
    
    5.

    Place the slide into a black 30-count slide rack.

    6.

    Pull the lever below the loading drawer on the autostainer to open.

    7.

    Place slide rack inside and close.

    8.

    Hit button to the left of loading drawer to begin staining run.

    9.

    When staining has finished, autostainer will periodically beep.

    10.

    Pull the lever below the exit drawer on the autostainer to open.

    11.

    Press button next to exit drawer so instrument knows the rack was removed. (Autostainer will

    continue to beep otherwise.)

    12.

    Coverslip and microscopically verify correct staining of control tissues.

    13.

    Denote onRHL Internal H&E Histologic Quality Check. If staining quality is suboptimal,

    troubleshoot.

    14.

    If staining is acceptable,write “PASS” on slide andstore in labeled control slide box next to

    autostainer.

    15.

    File form in H&E quality control binder at the end of the month.

    16.

    Any research slides for the day can be loaded in the same way once the review is complete.

    17.

    At the end of the workday, open cover and replace reagent lids to prevent evaporation. Close

    cover.

    18.

    Autostainer Program 1

    ABC
    1Xylene2 min
    2Xylene2 min
    3Xylene2 min
    4100% Alcohol2 min
    5100% Alcohol30 sec
    695% Alcohol2 min*
    795% Alcohol1 min
    Wash 5Water wash1 min
    8Hematoxylin*30 sec*
    Wash 4Water Wash30 sec
    Wash 3Water Wash2 min
    9Clearing Solution30 sec
    Wash 2Water Wash1 min
    10Bluing Solution30 sec
    Wash 1Water Wash30 sec
    1180% Alcohol30 sec
    12Eosin*30 sec
    1395% Alcohol30 sec
    1495% Alcohol30 sec
    15100% Alcohol30 sec
    16100% Alcohol30 sec
    17Xylene30 sec
    18Xylene30 sec
    Exit (19)XyleneHold-exit station

    *These are the only two stations with hard times. Slides may be held longer in other stations when multiple racks are being run at the same time.

    Protocol - Changing the Autostainer

    19.

    At the end of the work week, remove all staining containers and empty into the chemical waste containers located below autostainer

    20.

    Rinse containers out with water and, working over the sink, fill with a 10% bleach solution for at least 15 minutes or until visibly clean.

    21.

    Rinse out containers and place upright on a counter that has been covered with Wypall pads. Leave to air dry over the weekend.

    22.

    Fill water wash containers with 10% bleach and then 3 changes of waterto remove bleach residue (containers will automatically drain when filled).

    23.

    Flush 10% bleach solution followed with 3 changes of water through the base of the stainer. Use a bleach-soaked Wypall pad to spot clean any splashed staining reagents. NOTE: Manual H&E stain line should remain set up until autostainer is refilled and tested the next week to ensure one staining line is always available for use.

    24.

    At the beginning of the work week, refill staining containers and place back into autostainer.

    25.

    Reagent setup is as follows:

    ABCDEFGHIJKL
    XyleneXyleneXylene100% Alcohol100% Alcohol95% Alcohol95% AlcoholWash 1Wash 2Wash 3Wash 4Wash 5
    XyleneXyleneXylene100% Alcohol100% Alcohol95% Alcohol95% AlcoholEosin80% AlcoholBluing SolutionClearing SolutionHarris Hematoxylin
    26.

    Mark reagents changed on Autostainer Maintenance Log.

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