Hematoxylin and Eosin staining of Tissue Section

Procedure

  • Put the slide on hot plate/ flaming for few seconds ( to soften paraffin )
  • Then immediately dip slide in XYLENE for 5 min.(Minimum) to dissolve paraffin. After taking slide out, put some fresh xylene (Few drops) on the section to washout remaining paraffin.
  • Wash slide with 100% alcohol in slanting way over both side of slide to remove xylene
  • Then wash the slide with descending grade of alcohol ie, 90%,-70%- 50%-(2 min each), to hydrate the slide
  • Ultimately put the slide in water for 5 min
  • Bring up the slide from water and put in Koplins jar containing Harris’ Hematoxylene for 5 min. ( in modified method put hematoxylene on tissue section for 5 min)
  • Keep slide in rectangular koplin jar in running water for 5 min.
  • Dip 6the slide for 2/3 times in 1% acid alcohol (1% HCL),for differentiation (To remove excess stain in cytoplasm. colour  appear pink.
  • Keep the slide in running water again till it become blue colour (Colour of the nucleus)
  • Put 1% aquous eosin over slide for 2 min.
  • Dip in water for 2 min, (in running water) or more if staining is more.  ( 30 SEC. IN CASE OF SPIRITE EOSIN SOLUTION)
  • Then dehydrate slide by putting ascending grade of alcohol ie, 50%-70%-90%-Absolute alcohol  for 2 min each.
  • Dry in air.
  • Dip the slide in clean xylene for 1 min
  • Mount in Canada Balsam/DPX
For practical purpose, blotting is done by tissue paper in place of  sl. 12

SEE under MICROSCOPE.

H&E staining
H&E staining

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