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This study aims to develop an instrument that is used to assess scientific argumentation skills of prospective biology teacher students in microbiology practical activities. This research uses research and development methods which consist of define, design, develop and dissemination stages. This study produced a test instrument consisting of 5 essay questions each divided into 4 questions that measure aspects of argumentation, namely claims, data, warrant and backing. Each question number contains situations and questions related to the existence of microorganisms in daily life based on laboratory activities. The results of expert validation and trials show that the five questions are valid and reliable. Thus, the results of this study recommend the use of this test instrument for research that assesses the skills of scientific argumentation in microbiology lab activities.