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The page below is a sample from the LabCE course Histology Special Stains: Connective Tissue. Access the complete course and earn ASCLS P.A.C.E.-approved continuing education credits by subscribing online.

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Toluidine Blue Staining - Staining Protocol

Sample type required: Deparaffinized and rehydrated tissue section (3-5 microns) on positively (+) charged slides

Preferred fixative: 10% neutral buffered formalin (NBF)

Control: Skin

ReagentTimeTechnical Notes
Working toluidine blue

1 to 2 minutesSolution: 0.1g toluidine blue + 100 mL distilled water
Distilled water3 changes

Post staining procedure: Tissue section should be rinsed well in distilled water and then dehydrated with 95% and absolute alcohols. Follow with two changes of xylene and then coverslip.
Expected results:
  • Mast cells - Violet
  • Other tissue elements - Shades of blue