TITLE: U54 SCENT Histology H&amp;E Staining
AUTHORS: Shannon McCall

[DESCRIPTION]
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]
SECTION: 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.
SECTION: Protocol - Operating Autostainer and Running Daily Quality Control
2. Hit switch on the right side of autostainer to turn on and initialize.
SECTION: Protocol - Operating Autostainer and Running Daily Quality Control
3. Once initialization is complete, press button on screen to select staining program 1.
SECTION: Protocol - Operating Autostainer and Running Daily Quality Control
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
SECTION: Protocol - Operating Autostainer and Running Daily Quality Control
5. Place the slide into a black 30-count slide rack.
SECTION: Protocol - Operating Autostainer and Running Daily Quality Control
6. Pull the lever below the loading drawer on the autostainer to open.
SECTION: Protocol - Operating Autostainer and Running Daily Quality Control
7. Place slide rack inside and close.
SECTION: Protocol - Operating Autostainer and Running Daily Quality Control
8. Hit button to the left of loading drawer to begin staining run.
SECTION: Protocol - Operating Autostainer and Running Daily Quality Control
9. When staining has finished, autostainer will periodically beep.
SECTION: Protocol - Operating Autostainer and Running Daily Quality Control
10. Pull the lever below the exit drawer on the autostainer to open.
SECTION: Protocol - Operating Autostainer and Running Daily Quality Control
11. Press button next to exit drawer so instrument knows the rack was removed. (Autostainer will
continue to beep otherwise.)
SECTION: Protocol - Operating Autostainer and Running Daily Quality Control
12. Coverslip and microscopically verify correct staining of control tissues.
SECTION: Protocol - Operating Autostainer and Running Daily Quality Control
13. Denote onRHL Internal H&E Histologic Quality Check. If staining quality is suboptimal,
troubleshoot.
SECTION: Protocol - Operating Autostainer and Running Daily Quality Control
14. If staining is acceptable,write “PASS” on slide andstore in labeled control slide box next to
autostainer.
SECTION: Protocol - Operating Autostainer and Running Daily Quality Control
15. File form in H&E quality control binder at the end of the month.
SECTION: Protocol - Operating Autostainer and Running Daily Quality Control
16. Any research slides for the day can be loaded in the same way once the review is complete.

SECTION: Protocol - Operating Autostainer and Running Daily Quality Control
17. At the end of the workday, open cover and replace reagent lids to prevent evaporation. Close
cover.
SECTION: Protocol - Operating Autostainer and Running Daily Quality Control
18. Autostainer Program 1


 
*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.
SECTION: 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
SECTION: Protocol - Changing the 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.
SECTION: Protocol - Changing the Autostainer
21. Rinse out containers and place upright on a counter that has been covered with Wypall pads. Leave to air dry over the weekend.
SECTION: Protocol - Changing the Autostainer
22. Fill water wash containers with 10% bleach and then 3 changes of waterto remove bleach residue (containers will automatically drain when filled).
SECTION: Protocol - Changing the Autostainer
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.
SECTION: Protocol - Changing the Autostainer
24. At the beginning of the work week, refill staining containers and place back into autostainer. 
SECTION: Protocol - Changing the Autostainer
25. Reagent setup is as follows:  


 

SECTION: Protocol - Changing the Autostainer
26. Mark reagents changed on Autostainer Maintenance Log.