Cause of anxiety is very much widespread. Among different kinds of anxiety, academic anxiety is important anxiety type which generally encounters pupils. From various previous studies we can see almost all students are very easily caught by anxiety. Academic anxiety includes subject wise anxieties also. Biology is generally known as one of the most favourite subjects among students. Still, many of them are seen having anxiety in Biology. Chief aim of the present paper is construct and validate 'Biology Anxiety Scale'. Total 504 senior secondary students from India, took part in present study. Exploratory factor analysis and Confirmatory factor analysis were done to get the structural validity of scale with 252 sample size each. As a result, 3 dimensional and 20 items containing scale is generated. Three dimensions found were: biology test anxiety, biology class room and environment anxiety and biology laboratory anxiety. Alpha coefficient of the whole test found 0.935 and 0.896, 0.946 and 0.875 for each dimension