How to Change a Leica and Thermo Stainer

Laboratories worldwide rely on these popular automatic Stainer’s to stain large amounts of slides in an efficient manner so that lab technicians could focus on other tasks that can’t be automated. Each laboratory has their own protocol, programing, and process they utilize in their place of business. Today I am going to show step by step how I change the stainers I work with daily. 

The most common chemicals and reagents we are going to discuss below and replace are:

95% Alcohol 

100% Alcohol

Xylene

Bluing

Acid Alcohol

Periodic Acid Schiff’s

Hematoxylin

Eosin

Congo Red

Trichrome 

• Acid Alcohol, 95% Alcohol, and 100% Alcohol are used to neutralize and break down compounds and elements to give way for another stain to bind to the tissue. It is similar to a mosaic with multiple parts that make a whole masterpiece.

Put on PPE: Nitrile Gloves, Eye Protectant, mask or respirator. Note: The chemicals and reagents are toxic and harmful if inhaled. Lastly wear a lab coat to protect yourself and clothing for accidental spills and backsplash from the reagents and chemicals.  

Look at the schedule for which chemicals and reagents need changed based on the use, and the day of the week it’s on

Gather all of the chemicals and reagents needed to refill the pots in each stainer you change daily. 

Set up plastic containers with 2 funnels for the Alcohols and Xylene to be poured into. The Xylene container and funnel is red, and the alcohol container and funnel is white. Caution: DO NOT mix alcohol and xylene in the same container as it will cause a harmful chemical reaction.  Do not Pour Xylene down the drain, it is not safe for the environment.  

Open the front slide doors to access the pots of reagents

Locate the 95% Alcohol pots

Pour each 95% Alcohol into the white alcohol funnel attached to the white plastic container

Wipe out each container with a paper towel to remove residue, contaminates and water

Replace each 95% alcohol container back onto the Stainer and pour new 95% alcohol into the empty pots. Repeat until each pot is filled with new 95% Alcohol

Locate the 100% alcohol pots, and pour into alcohol container, wipe out and replace back in the Stainer. Pour 100% alcohol into empty pots. Note: Each pot has a proper place and location due to the programing of the Stainer. If a reagent pot was not put back in correct spot, the stain would not have the correct stain on it. This would hinder timely diagnosis.  

Locate Xylene pots and pour into Red Container and Funnel. Wipe out with a paper towel. Caution: Do not rinse out with water, it reacts to the oil base of Xylene. Xylene is used as a solvent to break down tissue to allow the stain to penetrate the tissue on the glass slides.  

Locate the Bluing Reagent pot. Pour down the sink with running water. Wipe out with a fresh paper towel. Replace the pot and pour the reagent labeled Bluing. Bluing is used to stain the Nucleolar chromatin and cell membranes of individual cells.

Locate the Periodic Acid Schiff’s Pot. Pour down the sink with running water. Wipe out with clean paper towel. Replace pot in proper place and refill with Periodic Acid Schiff’s. Periodic Acid Schiff’s (PAS stain) is commonly used to detect Renal disorders when glycogen storage disorder is being assessed.  

Image is normal PAS

Locate Hematoxylin pot, pour down the sing with running water. Wipe out with a clean paper towel. Pour fresh hematoxylin into clean pot. Hematoxylin is used to stain cell nucleus purple or dark blue.

Locate Eosin pot. Pour down the sink with running water. Pour new Eosin into pot and replace. Eosin is used to stain cell cytoplasm, collagen, red blood cells, and muscle fibers in a cell. It is most commonly used with Hematoxylin in a stain called H&E it has all of the elements of 2 separate stains to have a clearer picture of the cell structures in diagnosis. 

1st image is Normal H&E

2nd Image is H&E Minus Eosin

3rd image is H&E Minus Hematoxylin

Locate Congo Red pot. Pour down the sink with running water. Pour fresh Congo Red solution into empty pot. Replace back in proper location. Congo Red is used to detect certain type of protein (Amyloid) build up in organs. This build up could be indicative of Parkinson’s, Alzheimer’s, or one of over 36 other disorders

1st image normal congo red

2nd image is Congo red without the red

3rd image is congo red without the hematoxylin

Locate Trichrome reagent pot. Pour down the sink with running water. Pour fresh trichrome solution to empty pot. Replace back in proper location. Trichrome is used to differentiate between collagen and smooth muscle in tumors to most commonly detect cirrhosis of the liver

1st image normal trichrome

2nd image oversaturation of hematoxylin on trichrome

3rd image wrong reagent on trichrome

Obtain a control slide to test the process of changing the Stainer’s

Look at the control slides under a microscope to identify the proper staining process. This helps to verify a problem before multiple slides are stained improperly

Pour collected alcohol from white funnel and white container down the sink with running water. Wipe down funnel with paper towel and place at bottom of shelf for the next day’s Stainer changing. 

Collect Xylene container and pour into biohazard bin for proper disposal

Return used reagents to chemical storage cabinet. 

gather all used paper towels and throw into biohazard container

Remove all PPE and return to proper location in the laboratory

Remove gloves and throw away in biohazard container, and wash hands afterwards.