A new means of disaggregating cytology specimens in suspension using an immersible rotor device is described. The new rotor is compared to an automated syringing apparatus using cervical samples. Similar results using both devices are obtained for both normal and abnormal specimens.Key terms: Cytology, specimen preparation, automation A prerequisite for the (prelscreening of cytology specimens by an automated image analysis system is a reproducible and practical method for preparing smears of disaggregated cell suspensions. Many procedures have been described (1,4,10,14,16,17), but most are limited in that they are labor-intensive or very time-consuming. To make automated cytology an attractive addition to routine manual cytology, the specimen should be handled as little as possible, and the processing time should certainly not be much longer than the time needed to prepare a cervical smear with conventional manual methods.The Biological Precision Encoding and Pattern Recognition (BioPEPR) project (19) is concerned with full automation of cervical smear prescreening, including the preparation phase. In a previous paper (lo), we described a semiautomatic procedure by which about 100 cervical smears a day could be prepared. Because it proved difficult to fully automate this entire process as it was, certain steps in the process were reevaluated in order to make a fully automated system feasible.All of the automated disaggregation systems developed to date have been automated versions of the syringing procedure (1,4,7,9,10,16). With syringing it is difficult to fully automate the changing of the needles or the syringes, and to provide adequate rinsing of the device between samples to prevent specimen contamination.The present paper describes a new method of disaggregating cell suspensions. Cell disaggregation with this device, a specially designed rotor, is easy to automate and has proved to be efficient. In this paper the rotor is described, and results obtained with this rotor when disaggregating cervical specimens are presented. MATERIALS AND METHODSCervical cells were collected as second or third scrapes using a plastic spatula that was rinsed in a preservative phosphate-buffered saline solution, containing 20% ethanol. Disaggregation was performed either by a syringing technique using a peristaltic pump device (10) for 15 min with 19-gauge needles and a speed of 55 mV min, or by a rotor immersed in the sample vial containing 9 ml of cell suspension (Fig. 1A). For comparative studies the original sample was divided before disaggregation into two to four sample containers, and the fluid volume was adjusted back to 9 ml.After disaggregation the cells were deposited onto glass microscope slides, as described elsewhere (10).The rotor device used consists of a 24-V DC Maxon motor (Sachseln, Switzerland) with a vertical axle on which different cylindrical rotor heads can be mounted. The distance from the outer edge of the rotor head to the vessel wall is about 2 mm. Figure 1B shows two examples of hollow rotor heads,...
This paper describes a new automated system to prepare slides of cytological material from suspension. The system collects material on a filter tape by filtration and transfers it to glass slides by means of pressure-fixation. Using cervical cells as a model, results show that a well-defined cell number is evenly deposited over a standardized area, while a small number of cells is retained on the tape and a negligible number lost in the filtrate. Contamination is very small. Application of the system to other cytological material (fine needle aspirations, monolayer and cell suspension cultures, agar cultures, and isolated nuclei) is shown. In general, more than one slide can be made from one sample. Several histological staining procedures as well as immunofluorescence labeling protocols can be applied to the preparations obtained in this way. This system thus introduces a method that will standardize specimen preparation, is quick, saves operator time, and can be used for both diagnostic and research applications.Key terms: Specimen preparation, cervical cells, fine needle aspirations, cell cultures, tumor cell colonies, isolated nuclei Light microscopical examination of cellular material by human observers has been performed almost exclusively on conventionally prepared and stained slides. Demands on the quality of the preparations were not so extreme because the observer could correct for most preparative shortcomings. To improve this visual and qualitative evaluation, new techniques for cell interpretation are being developed. Our laboratory is involved in this research, developing, among others, specific immunohistochemical stains to detect the tissue origin of tumor cells (18)(19)(20)(21), to detect quantitative morphologic and cytochemical changes using image analysis derived techniques (6,30,31), and to assess the malignant behavior of tumor cells in an in vitro model system (5,8,9,27). These new techniques require the preparation of reproducible, well-standardized specimens with homogeneously dispersed, nonoverlapping, single cells or cell groups. Therefore, to complement this research, the development of new cytopreparatory techniques has begun (12,15,16). One of the steps in this specimen preparation procedure is the deposition of cells from suspension onto glass slides. Numerous approaches have been described to achieve this goal. These approaches can be subdivided into three classes: centrifugation (1,2,10,24-261, sedimentation (7,12,28), and transferring the cells through an intermediate filter onto a slide (2,11,16,17,22,23). In the last procedure, the cells either remain on the filter (2,17) or are transferred to the slide by touch (22,23) or pressure (11,161. In a previous paper (16) we have evaluated the latter method. Cells were collected on a polycarbonate filter membrane and transferred to glass slides by simultaneous pressure-fixation. This procedure proved to be relatively robust with respect to parameters that might influence cell recovery such as filtration rate, filter pore size, and pr...
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