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Optimizing fixation and staining to reduce cell detachment in wet-fixed direct smears, a quantitative evidence-based quality improvement study

Year: 2026
Type: article
Abstract: Wet-fixed cytologic smears are inherently prone to cell detachment, which causes cross-contamination and reduces smear cellularity, potentially leading to false-positive diagnoses and diagnostic uncertainty. This study evaluated the effects of the intermediate alcohol step, fixation duration, and slide type on detachment to establish an evidence-based optimized protocol. We evaluated: (i) the necessity and optimal concentration of the intermediate alcohol step; (ii) fixation duration from 30 min to 24 h; and (iii) slide type (ordinary vs. positively charged). Detachment was scored semi-quantitatively using a streak-wiping method. Wilcoxon and Friedman tests were used for paired and block comparisons, respectively. The intermediate alcohol step significantly reduced detachment ( p < 0.05), with 50% ethanol superior to 75% ( p = 0.012). Prolonged fixation progressively decreased detachment ( p < 0.001), with no further statistically significant reduction beyond 3 h. Under optimized conditions, ordinary and positively charged slides performed comparably ( p = 0.811). An evidence-based strategy for laboratory quality improvement—extending fixation toward 3 h, including an intermediate alcohol step (preferably 50% ethanol) at each transition between water and 95% ethanol, and using ordinary slides—substantially reduces detachment. By preserving cellularity and helping to reduce the risk of cross-contamination, this protocol offers an implementable quality improvement tool for cytology laboratories, potentially enhancing diagnostic reliability and patient safety in wet-fixed cytologic preparations.
DOI: 10.1038/s41598-026-76374-3
Language: English
Open Access status: gold
Estimated APC paid: $2,690
OpenAlex ID: W7222836214
FWCI:
Cites: 15
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Related to: 10