Background: Acupuncture is now applied for treating numerous conditions. The safety was pay more and more attention in the worldwide. Thus, it is important to establish a reference for safe needling depth of acupoints by nowadays image exam. In this study, we evaluated the depths to which acupuncture needles can be inserted safely in the chest and back acupoints. We also investigated the variations in safe depth according to sex and body mass index (BMI). Methods: We retrospectively studied computed tomography (CT) images of adult patients who underwent chest CT at China Medical University Hospital between January 2002 and January 2016. The safe depths of chest and back acupoints were measured directly from the CT images. The relationships between the safe depths of these acupoints and sex and BMI were analyzed. Results: Totally 222 patients were enrolled in this study, including 105 males and 117 females. The results indicated significant differences in safe needling depth between males and females in chest acupoints KI22, KI23, SP18, SP19, PC1, CV21, CV22 and back acupoints GV10, GV11, BL17, BL45, GB21 (p < 0.05). Safe needling depth of chest acupoints KI27, KI23, ST15, ST16, ST18, SP17, SP18, SP19, SP20, PC1, CV19 and back acupoints GV14, BL13, BL43, BL46 differed significantly in different BMI groups (p < 0.05). Generalized linear regression model revealed that BMI was the most crucial factor in determining the safe depth. Conclusions: BMI are relevant factors in determining the safe needling depths of chest and back acupoints. In order to prevent complication of acupuncture, physicians should consider wide variations in needle depth when performing acupuncture.