SU DAN III OIL RED-O

SU DAN III OIL RED-O

Lipids that exists as fats, have an affinity for the Sudan dyes. The adsorbing power of the fats is related to the dye concentration, temperature and the physical state of the fats. –

Sections
Cryostat sections are used for staining, since the fat would dissolve in the paraffin block sections.

Solutions

1) Saturated solution of Sudan III, oil red        -        O in 70% alcohol.
2) Haematoxylin


Procedure - Take 2 sections.
1)   Air dry sections and rinse in 70% alcohol.
2)   Stain with Sudan fed soil red for 10-20 mts.
3)   Rinse in 70% alcohol.
4)   Wash in water. Stain one section with Haematoxylin for 30 secs.
5)   Rinse in water and mount in glycerine jelly.
6)   Seal the edges of the coverslip with either molten wax or nail polish.


Result
                   Lipids           -        Glistening red.
                   Nuclei           -       Blue.





Glycerine Jelly

                        Gelatine                       -                      10gms
                         Distilled water            -                       60ml
                        Glycerol                       -                      70 ml
                         Phenol                        -                      250mg

Dissolve the gelatine in distilled water in incubator or water bath before adding the remaining ingredients. Then medium sets solid and requires melting before use.


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