Stain Profile
PTAH
Phosphotungstic acid hematoxylin
Purpose, principle, optimal fixative, & procedure
From Self Instruction textbook, notes & etc
Purose: Method of Mallory used to demonstrate the cross-striations arrangements of actin & myosin contractile proteins, and also fibrin in skeletal muscle. The cross-striations are a diagnostics feat of rhabdomyosarcomas (et other skeletal muscle diseases). Can be used to demo glial fibers & myelin. Outdated because of IHC.
Principle: PTA to hematein 20:1, tungsten bins to all available hematein to give blue lake. Metal-hematein lake stains blue (cross-strations, fibrin & nuclei) and PTA stains red-brown (collagen). A polychrom stain.
Fixative: Zenker>NBF
Stain Procedure
Phosphotungstic acid hematoxylin (PTAH)
From Self Instruction textbook, notes & etc
- Deparaffinize sections & hydrate to distilled water.
2.Formalin fixed, then mordant in Zenker with 5% acetic overnight
3.Rinse in tap
4.Place in Gram iodine for 15min
5.Rinse in tap
6.Place in 5% aqueous sodium thiosulface 3 min (mercury removal)
7.Wash in tap for 10min
8.Place sections in 0.25% potassum permanganete for 5min
9.Rinse in tap
10.PLace in 5% oxcalic acid for 1min
11.Wash in running tap 10 min
12.Stain in PTAH solution overnight
13.Dehydrate quickly in 2x 95% & 100% ROH, clear in xylene & then mount with synthetic resin
Remember Zenker contains mercuric chloride so the steps with gram iodine & sodium thiosulfate are critical to avoid fixation pigment
