Sputum induction is an investigative method to stimulate the natural secretion of mucus from the airways. The substance obtained in this process is called induced sputum. It originates from the lungs or lower parts of the airways. The induced sputum is often gathered by inhalation of aerosols of the hypertonic saline solution produced by an ultrasonic nebulizer. Drops of saline irritate the mucous membranes, trigger coughing and cause expectoration of sputum. Appropriate and precise pre-preparation and processing of given samples for analysis are of key importance, because only in this way do we get quality cytological compounds that help us assess the functioning of the lungs and inflammation in the airways.
In the dissertation, we compared two methods of pre-preparation and processing of taken samples of induced sputum. We have been using the first method for a long time in routine practice and is hence standardized. However, the second method is newer and has not yet been established for work in clinical practice. Using the recently developed approach, we optimized four factors, the initial mass of the samples of the induced sputum - plaques, temperature and time conditions of work and the consumption of the fundamental reagent. Among the methods, we compared cell viability, total cell concentration and the proportion of individual cell types on cytological preparations. We were interested in whether the newer procedure, despite the change in the ten steps of the protocol, gives reliable and comparable results with the existing method and whether certain advantages would facilitate work in this field in the future and replace the already standardized procedure.
The results showed that the newer method, despite a statistically significant lower mass of samples of induced sputum, gives completely comparable results with the routine method in terms of cell profile. Slightly lower cell viability was present in only four samples that deviated from the majority, while the overall cell concentration in the new method was statistically higher. Although the new procedure has the additional centrifugation step, we still shorten the time from the acceptance of samples in the laboratory to the production of permanent cytological preparations. That is particularly noticeable when there is a large number and samples of poorer quality. Using the new process, we also reduced the consumption of the main and most expensive reagent, which is an essential link in both methods of processing samples of induced sputum, as it reduces the presence of mucus, improves homogenization and the dispersion of cells. With its lower consumption, we significantly increase the economic efficiency of the method, and at the same time, the results are comparable to the already existing procedure.
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