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Pre-experimental studies on the development of three tier multiple choice tests were conducted to detect and measure of students tree thinking in learning concept of classification. Three tier multiple choice test was developed with web integration of a Google Classroom application for easy feedback. The study with the one group pretest-posttest design involved a number of high school students (n = 29) who had studied the concept of Animal Classification. The research procedure was carried out through stages (literature study, problem analysis, and instrument design). The instrument was, then, validated by expert and analysed with ANATES version 4.0. Tree thinking was measured on the basis of identification of evolutionary characters, phylogenetic relationships, concept of clad, MRCA (Most Recent Common Ancestor), sister groups, tree topologies, number of evolutionary changes, and sequence of evolution in species. The validation results showed that the three tier multiple choice test can be used to measure of students tree thinking. In addition, the results analysis of items with ANATES version 4.0 indicated that the test has high reliability (r = 0.69), very good of differences (D> 0.4) on 12 items, and very good of difficult on 14 items. In conclusion, this study showed that three tier tests for measuring high school student-s tree thinking consist of 20 test items with double MC (4 options each for answer and reason), with 11 test items were received directly (60%), 3 items were received but with repairment (15%), 4 items were rejected (25%).