Toluidine Blue staining

Standard staining  for acrylate

·        Deplasticizer I - III                          each 5 min.                              

·        2´Ethanol 100%                              each 2 min.                              

·        Ethanol 96%                                   2 min.

·        Ethanol 80%                                   2 min.

·        Ethanol 70%                                   2 min.

·        Ethanol 50%                                   2 min.

·        Aqua dest                                       wash by submerging 3-5 times

·         Toluidin blue 1%, pH 4,5                30 min.

·         Aqua dest                                      wash

·         Ethanol 50%                                  wash

·         Ethanol 70%                                  2 min.

·        Ethanol 80%                                   2 min.

·        Ethanol 96%                                   2 min.

·        Ethanol 100%                                 5 min.

·        Ethanol 100%                                 wash

·        Xylol I - III                                      each 5 min.

·        Mount with DPX

Solutions:

1g Toluidin blue-O per 100ml aqua dest.

Adjust pH to 4,5 with 0,1% NaOH or 0,1% HCl

Store at 4°C

Staining results:

darkblue:          Nucleus

blue:                 osteoblassts

turquois:           osteoclasts

light blue:         mineralized bone (with darker cement lines)

purple:              cartilage