Technical pitfalls in middle ear gas studies: Errors introduced by the gas permeability of tubing and additional dead space

Conclusions. We demonstrated errors introduced by the gas permeability of the tubing and additional dead space. Materials with practically no permeability, such as glass, must be used to overcome the loss or gain of gas through the walls of tubes used for studying ME gas variations. Experiments conducted at a constant volume (variable pressure) require the smallest possible tubing volume in order to obtain good sensitivity and improve the accuracy of the results. Objectives. Experimental studies that investigate middle ear (ME) gas exchanges, using either pressure measurements or volume changes, are conducted using various tubing connections between the ME and a measuring device. The aims of this study were to highlight experimental problems due to the errors introduced by (i) the gas permeability of the tubing used and (ii) additional dead space in experiments conducted at constant volume. Material and methods. The problem of the gas permeability of the tubing was addressed by comparing three types of tube (silicone, polyethylene, PVC) with a glass tube. Horizontally placed tubes were connected to a syringe filled with pure CO ₂ via a gas-tight valve. The end of each tube tested was plunged into colored water (5% Coomassie Brilliant Blue R-250). The tube was washed out with CO₂ until gas bubbles were seen leaving it. The valve was then closed and the movement of water in the tube was observed. The same experiments were repeated for pure O₂. The problem of the error introduced by the additional dead space volume was addressed at a theoretical level using the well-known gas laws. Two conditions were studied: condition A, in which the experiment was conducted at constant pressure and the volume was measured; and condition B, in which the experiment was conducted at constant volume and the pressure was measured. The main outcome measure of each condition was the calculated variation in the final number of moles of gas involved. Results. No water movement was observed in glass tubes. In contrast, plastic tubes exhibited significant gas permeability effects for both CO₂ and O₂. The colored solution advanced at a faster rate with CO₂ than O ₂ but differently for each type of tubing. For condition A, gas transfer was independent of the volume of the external tubing and was accurately measured by the displacement of the droplet in the lumen. In contrast, for condition B, the pressure variations were influenced by the volume of the tubing.