U54 SCENT Histology H&E Staining

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

right side of autostainer. Close cover.

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

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

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

a. H&E QA

b. Date

c. Tech initials

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

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

Place slide rack inside and close.

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

When staining has finished, autostainer will periodically beep.

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

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

continue to beep otherwise.)

Coverslip and microscopically verify correct staining of control tissues.

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

troubleshoot.

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

autostainer.

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

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

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

cover.

Autostainer Program 1

A	B	C
1	Xylene	2 min
2	Xylene	2 min
3	Xylene	2 min
4	100% Alcohol	2 min
5	100% Alcohol	30 sec
6	95% Alcohol	2 min*
7	95% Alcohol	1 min
Wash 5	Water wash	1 min
8	Hematoxylin*	30 sec*
Wash 4	Water Wash	30 sec
Wash 3	Water Wash	2 min
9	Clearing Solution	30 sec
Wash 2	Water Wash	1 min
10	Bluing Solution	30 sec
Wash 1	Water Wash	30 sec
11	80% Alcohol	30 sec
12	Eosin*	30 sec
13	95% Alcohol	30 sec
14	95% Alcohol	30 sec
15	100% Alcohol	30 sec
16	100% Alcohol	30 sec
17	Xylene	30 sec
18	Xylene	30 sec
Exit (19)	Xylene	Hold-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.

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

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.

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

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

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.

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

Reagent setup is as follows:

A	B	C	D	E	F	G	H	I	J	K	L
Xylene	Xylene	Xylene	100% Alcohol	100% Alcohol	95% Alcohol	95% Alcohol	Wash 1	Wash 2	Wash 3	Wash 4	Wash 5
Xylene	Xylene	Xylene	100% Alcohol	100% Alcohol	95% Alcohol	95% Alcohol	Eosin	80% Alcohol	Bluing Solution	Clearing Solution	Harris Hematoxylin

Mark reagents changed on Autostainer Maintenance Log.