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After 30 min, a 100 L of acidic stop solution was added into each well and the absorbance at 450 nm was recorded using a using a Biotek Synergy Neo2 HTS Multi-Mode Microplate Reader (Mnnedorf, Switzerland)

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Such was the case in this study, that the numbers of microglia in alcoholic brain were higher than those in mice given ALC plus alcohol or in the pair-fed controls