How to operate phospholipid iron hematoxylin (FEH)_Industrial additives

Background and overview^[1]

Phospholipid iron hematoxylin (FEH) staining solution is used for phospholipid staining. Phospholipids refer to lipids containing phosphoric acid and are complex lipids. Phospholipids are components of biological membranes and are divided into two categories: glycerophospholipids and sphingomyelin. Phospholipids are amphiphilic molecules with a hydrophilic nitrogen- or phosphorus-containing head at one end and a hydrophobic (lipophilic) long hydrocarbon chain at the other end. Elleder found that phospholipids can be revealed by ferric hematoxylin, which is simpler, faster, and more sensitive than DAH. The disadvantage is that the staining effect is good only after the tissue is defatted with acetone, and sometimes the cell nuclei and lipids are stained black and blue at the same time.

Composition^[1]

Operation steps (for reference only) ^[1]

1. Soak one slice in chloroform:methanol solution (1:1) at room temperature for 1 hour as a control, and soak another slice in anhydrous acetone for 15 minutes at 4°C.

2. Fix in FeH fixative for 30 minutes.

3. Rinse with distilled water.

4. Prepare FeHstain in advance: for every 5ml of hematoxylin solvent, add 0.1g of FeH hematoxylin. After fully dissolved, add 25ml FeHbuffer, which is FeHstain.

5. Add FeHstain for staining for 7 to 10 minutes.

6. Rinse with distilled water.

7. After diluting the FeH differentiation solution with an equal amount of distilled water, put the slices into the diluted FeH differentiation solution and soak the organic bentonite rheology additive several times.

8. Rinse with tap water.

9. Dehydrate with acetone, make it transparent with xylene, and mount with DPX mounting medium.

Notes^[1]

1. Phospholipids dissolve easily, so the tissue should be fixed immediately after removal, otherwise it will be difficult to stain.

2. This staining solution has a better staining effect on frozen sections.

3. Stained specimens must be stored away from light, otherwise they will fade easily.

4. For your safety and health, please wear a lab coat and disposable gloves.

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