Melanin bleaching with dilute hydrogen peroxide: a simple and rapid method.
Melanins are naturally occurring pigments in both normal and pathologic tissues. Two common bleaching processes are potassium permanganate followed by oxalic acid treatment and dilute hydrogen peroxide (H2O2) process. The potassium permanganate/oxalic acid method is faster and more easily incorporated in conventional daily immunostaining protocols, whereas the dilute H2O2 method requires 24 hours. This study aimed to reduce melanin bleaching time by using a 10% H2O2 dilution. First, reaction time was reduced to 30 minutes by raising the temperature to 65°C. Second, containers with high thermal conductivity were used to improve bleaching effectiveness. Experimental comparisons of melanin treatments with H2O2 contained in an iron jar, a glass coplin jar, and a plastic steel jar obtained bleaching time of 20, 30, and 40 minutes, respectively. These modifications of the conventional bleaching method significantly improve the speed and efficiency of the procedure and are recommended when performing immunohistochemical studies.
Department of Pathology, Kaohsiung Medical University Hospital.
Chia-Hsing
CH
Liu
Chih-Hung
CH
Lin
Min-Jan
MJ
Tsai
Wan-Tzu
WT
Chen
Chee-Yin
CY
Chai
Ya-Chun
YC
Huang
Kun-Bow
KB
Tsai
D057886:Bleaching Agents / Q000737:chemistry*; D004801:Eosine Yellowish-(YS); D006416:Hematoxylin; D006801:Humans; D006861:Hydrogen Peroxide / Q000737:chemistry*; D007150:Immunohistochemistry / Q000191:economics / Q000379:methods* / Q000592:standards; D008543:Melanins / Q000737:chemistry*; D019815:Oxalic Acid / Q000737:chemistry; D011196:Potassium Permanganate / Q000737:chemistry; D013048:Specimen Handling; D013696:Temperature
D057886:Bleaching Agents; D008543:Melanins; D011196:Potassium Permanganate; D019815:Oxalic Acid; D006861:Hydrogen Peroxide; D004801:Eosine Yellowish-(YS); D006416:Hematoxylin