AbstractThis study investigates the rule that repeating cytological preparations on liquid‐based cytology improves sample adequacy, diagnosis, microbiological, and hormonal evaluations. We reviewed 156 cases of pap‐stained preparations of exfoliated cervical cells in two slides processed by DNA‐Cytoliq System®. After sample repeat/dilution, limiting factors affecting sample adequacy were removed in nine cases and three unsatisfactory cases were reclassified as satisfactory. Diagnosis was altered in 24 cases. Of these, the original diagnosis in 15 was atypical squamous cells of undetermined significance; after the second slide examination, diagnosis in 5 of the 15 cases changed to low‐grade squamous intraepithelial lesion, 3 to high‐grade squamous intraepithelial lesion, and 7 to absence of lesion. Microbiological evaluation was altered, with Candida sp. detected in two repeated slides. Repeat slide preparation or dilution of residual samples enhances cytological diagnosis and decreases effects of limiting factors in manually processed DIGENE DCS LBC. Diagn. Cytopathol. 2007;35:663–669. © 2007 Wiley‐Liss, Inc.