Characterizing the Effect of Processing Technique and Solution Type on Cytomorphology Using Liquid-Based Cytology

Abstract

<b><i>Introduction:</i></b> Liquid-based cytology (LBC) is increasingly used for nongynecologic applications. However, the cytological preparation of LBC specimens is influenced by the processing technique and the preservative used. In this study, the influence of the processing techniques and preservatives on cell morphology was examined mathematically and statistically. <b><i>Methods:</i></b> Cytological specimens were prepared using the ThinPrep (TP), SurePath (SP), and AutoSmear methods, with 5 different preservative solutions. The cytoplasmic and nuclear areas of Papanicolaou-stained specimens were measured for all samples. <b><i>Results:</i></b> The cytoplasmic and nuclear areas were smaller in cells prepared using the 2 LBC methods, compared to that prepared using the AutoSmear method, irrespective of the preservative used. The cytoplasmic and nuclear areas of cells prepared using the SP method were smaller than those of cells prepared using the TP method, irrespective of the preservative used. There were fewer differences among the cytoplasmic areas of cells prepared with different preservative solutions using the TP method; however, the cytoplasmic areas of cells prepared using the SP method changed with the preservative solution used. <b><i>Conclusions:</i></b> The most significant difference affecting the cytoplasmic and nuclear areas was the processing technique. The TP method increased the cytoplasmic and nuclear areas, while the methanol-based PreservCyt solution enabled the highest enlargement of the cell. LBC is a superior preparation technique for standardization of the specimens. Our results offer a better understanding of methods suitable for specimen preparation for developing precision AI-based diagnosis in cytology.

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“…The LBC method, which does not depend on the technique used by technicians for preparation, is an excellent processing technique for advancing quality control and standardization of cytological preparation. However, the cytologic forms differ depending on the LBC preservative solution [18], and even in this study, in which the same cells were treated with different preservative solutions, the cytologic features were different (Fig. 1).…”

“…The nuclear darkness of the SurePath specimens was greater than that of ThinPrep [32]. In a previous study, we analyzed the cytological form mathematically and statistically using the same LBC preservative solution as this study, and the ThinPrep method not only has a significantly larger cytoplasmic and nuclear area than the SurePath method; even with the same processing technique, the Pre solution has larger cells than those of the other solutions [18]. Various cytologic forms were observed in the Pre solution specimen, which were neither shrunken nor rounded.…”

“…The cells were divided into five groups. The samples were centrifuged at 600g for 5 min at 25 ° C, and CytoRich TM Blue (Blue) (Becton, Dickinson and Company, Franklin Lakes, NJ, USA), CytoRich TM Red (Red) (Becton, Dickinson and Company), CytoCollect (CyC) (Muto Pure Chemicals Co., Ltd., Tokyo, Japan), PreservCyt ® (Pre) (Hologic Inc., Marlborough, MA, USA), or TACAS TM Ruby (TAC) (Medical & Biological Laboratories Co., Ltd., Nagoya, Japan) were added to each sediment [18]. Liquid-based preparations were fixed overnight and prepared using a ThinPrep 5000 Processor (Hologic Inc.).…”

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“…The ThinPrep method uses a ThinPrep 5,000 processor to gently disperse the cell suspension and homogenize the cell population. The cells are then automatically collected on disposable polycarbonate filters and transferred to glass slides within a 20 mm diameter circle ( 18 ). In the SurePath method, centrifugation is used to attach the cell pellets to the glass slides by gravitational sedimentation and electrical adhesion ( 18 ).…”

“…The cells are then automatically collected on disposable polycarbonate filters and transferred to glass slides within a 20 mm diameter circle ( 18 ). In the SurePath method, centrifugation is used to attach the cell pellets to the glass slides by gravitational sedimentation and electrical adhesion ( 18 ). There are differences in cell morphology, background, and artifacts between the ThinPrep and SurePath methods ( 19 , 20 ).…”