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Within a week, i.e., on 4 th day, micronuclei appeared along with growth of bacterial colonies invading from periphery toward the center.
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In 24 hours, the cytoplasmic vacuoles increased in number and size. The group 1 cytosmear kept at room temperature showed cytoplasmic changes in 2 hours along with small vacuoles and patchy distribution of Pap stain, as shown in Figure 1a. Cellular features found in degenerative change which were chosen for the assessment were contaminants, bacteria and fungi, cytoplasmic inclusions, disintegrating cytoplasm, vacuolated cytoplasm, and presence of karyolysis, karyorrhexis, and pyknosis.
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Cytological assessment of degeneration was done by two screeners in a blinded fashion (without knowing the time of delay for each preparation). Smears were initially examined under light microscope (Olympus BX41 digital microscope) under 40X and 100X magnifications. Group 1 consisted of cytosmear in which fixation was delayed and Group 2 with addition of external factors. Broadly, the study was divided into two main groups as discussed in Tables Tables1 1 and and2. All investigations on the subjects included an informed consent and patient's anonymity is preserved.įrom every six smears prepared, two control smears were immediately stained following 15-minute fixation, whereas two each were stained with standard Papanicolaou (Pap) and H and E and were used in the study. The prospective study included six (06) smears prepared each from the buccal mucosa of 66 healthy volunteers in the age range of 20 to 30 years (mean age, 24 years) with no history of oral and/or systemic diseases such as diabetes, malignancy, anemia, etc. This study was conducted with two-fold objectives first, to observe changes in cells caused by delayed fixation/addition of external factors and second, to determine the impact of these mistakes on scientific interpretation and/or diagnosis.Ĭytological study of oral cells is a nonaggressive technique that is well accepted by the patient. Similarly, the importance of identifying an extraneous factor induced during smear processing, from pathogens and pathologists, needs recognition. To achieve this, a minimum of 15-minute immediate fixation prior to staining is essential. Thus, rapid fixation of smears is necessary to preserve cytologic details of cells. Regardless of the collection technique, specimen obtained from the mouth should be processed immediately. The duration between collection and preparation of the sample before cellular damage occurs depends on pH, protein content, enzymatic activity, and the presence or absence of bacteria. There are many factors which can affect a cytosmear unintentionally by a qualified health personnel or due to unawareness of an auxiliary staff. Mishandling, delay in processing, or injudicious contamination might lead to misinterpretation, flawed diagnosis, or false-positive/negative results. Cytology, if performed with accuracy, can play an important role in early diagnosis. The importance of art and science of cytosmear making and its proper handling is thus imperative. Although an adjunct to the gold standard histology, cytology is a very commonly performed procedure in clinical practice. These have been well studied at the tissue level but not so well at the cytological level.Ĭytology is a simple, quick, non-invasive method for tissue analysis of shed cells from an epithelial surface. Artifacts can interfere with histological assessment by changing the tissues appearance, mimicking other known tissues, creating confusion due to their unidentifiable structure, and/or hiding histological landmarks. These are not present in normal tissue and generally arise on a surgeon's chair or pathologist's laboratory from outside sources. Artifacts are structures or features which are being produced by the processing of a tissue, and may interfere with normal histological interpretation and/or diagnosis.