Large Language Models (LLMs) have become foundational in modern language-driven applications, profoundly influencing daily life. A critical technique in leveraging their potential is role-playing, where LLMs simulate diverse roles to enhance their real-world utility. However, while research has highlighted the presence of social biases in LLM outputs, it remains unclear whether and to what extent these biases emerge during role-playing scenarios. In this paper, we introduce BiasLens, a fairness testing framework designed to systematically expose biases in LLMs during role-playing. Our approach uses LLMs to generate 550 social roles across a comprehensive set of 11 demographic attributes, producing 33,000 role-specific questions targeting various forms of bias. These questions, spanning Yes/No, multiple-choice, and open-ended formats, are designed to prompt LLMs to adopt specific roles and respond accordingly. We employ a combination of rule-based and LLM-based strategies to identify biased responses, rigorously validated through human evaluation. Using the generated questions as the benchmark, we conduct extensive evaluations of six advanced LLMs released by OpenAI, Mistral AI, Meta, Alibaba, and DeepSeek. Our benchmark reveals 72,716 biased responses across the studied LLMs, with individual models yielding between 7,754 and 16,963 biased responses, underscoring the prevalence of bias in role-playing contexts. To support future research, we have publicly released the benchmark, along with all scripts and experimental results.