Lendrum, Slidders & Fraser's Alcian Blue
for amyloid
Materials
- Stock A
Alcian blue 8GX 1 g Ethanol 80% 100 mL - Stock B
Sodium sulfate, hydrate 1 g Distilled water 100 mL - Working solution
Stock A 45 mL Stock B 45 mL Glacial acetic acid 10 mL Mix. Leave 30 minutes. Prepare daily.
- Acetic ethanol
Ethanol, 95% 45 mL Distilled water 45 mL Glacial acetic acid 10 mL Prepare daily.
- Borax Ethanol
Borax as required Ethanol, 80% 100 mL Dissolve borax to saturation.
Tissue Sample
Paraffin sections of formalin fixed tissues are satisfactory. If using the more complex trichrome counterstains, formal sublimate fixation is preferred.
Protocol
- Bring sections to water via xylene and ethanol.
- Place into acetic alcohol for 1 – 2 minutes.
- Place into the working solution for 2 hours.
- Place into acetic alcohol for 1 – 2 minutes.
- Wash with water.
- Place into alcoholic borax for 30 minutes.
- Wash with water.
- Counterstain (see notes).
- Dehydrate with absolute ethanol.
- Clear with xylene and mount with a resinous medium.
Expected Results
- Amyloid – blue green
- Other tissue – according to the counterstain
Notes
- The simplest counterstain is the van Gieson.
- If counterstaining with PAS, treatment with alcoholic borax may be omitted.
- If counterstaining with silver impregnation for reticulin, the method must not include a Mallory bleach.
- The authors suggested their own, more complex trichrome counterstain.
Safety Note
Prior to handling any chemical, consult the Safety Data Sheet (SDS) for proper handling and safety precautions.
References
- Lendrum, A.C., Slidders, W. and Fraser, S., (1972),
Renal hyalin: A study of amyloidosis and diabetic fibrinous vasculosis with new staining methods.,
Journal of Clinical Pathology, v 25, page 373.