Heo et al[23] also randomized patients with large stones (over 15 mm in diameter) in an ESLBD group (12 mm to 20 mm balloon diameter, dilation time 60 s) and in an EST group. When EST was performed alone, it was completed to its full length (major EST), whereas combined with dilation it stopped after reaching one third of the full length that could be theoretically reached (minor EST). The reported successful stone removal was 94.4% for the ESLBD group and 96.7% for the EST group, while complication rates were similar between the 2 groups (5% vs 7% respectively). Pancreatitis and cholangitis appeared in the same proportion (4% and 1% respectively). ML for stone extraction after failure of the conventional methods was required in 8% of the ESLBD group and in 9% of the EST group. Based on the similar rates of successful stone removal and complications, the analysis suggested ESLBD as an alternative option in large stone endoscopic treatment.
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Draganov et al[27] evaluated the efficacy and safety of ESLBD in patients with difficult stones who had failed stone extraction with standard techniques after full length EST. Successful complete stone removal was achieved in 95% of the patients, while in 84% the stone clearance was accomplished without additional ML. Mild complications occurred in 6% of the cases.
Although EST or ESLBD have been performed for removal of bile duct stones in patients with Billroth II gastrectomy, the reported results are not completely satisfactory. In a recent trial, a Korean group performed stone extraction after limited EST followed by dilation up to 15 mm. ML was required in 11.5% of the cases, while in all cases stones were successfully removed in a maximum of three consecutive sessions without significant complications (bleeding, pancreatitis or perforation). The authors consider ESLBD as an effective and safe method of stone removal in patients with Billroth II gastrectomy[73].
Critical point drying (CPD), a standard method used during sample preparation in scanning electron microscopy, has also been successfully applied to enhance micro-CT imaging of soft-tissue samples by the elimination of liquids (20,25,26).
The applied contrast agents have been used primarily because of their relative ease of use and low toxicity (16,18) while osmium has been used successfully for contrasting eyes in previous studies (12,14). In particular, iodine-containing solutions have proven successful for contrasting ocular tissue (13-15) and was confirmed with this study. In contrast to the present work, previous study using PTA to contrast eyes did not provide corresponding visualizations (14). PTA is known for its low penetration rate (16) and was demonstrated here particularly on the lens. The 30 weeks applied in our study exceeded by far the incubation times on previous approaches [96 h, see (14)]. For the visualization of the lens, iodine based contrast ag ents are more suitable than PTA for which the incubation times of more than 30 weeks applied here was still insufficient.
Under moist conditions, micro-CT scans of untreated samples (M-UT) generally possess low differential contrast between tissues which complicates a proper analysis (Figure 2A). In order to overcome this problem in soft tissue samples, a number of contrast media have been successfully tested and applied to enhance radiodensity (16,17). With these methods of contrasting and CPD, the necessary prerequisites are created to visualize delicate morphological structures of the eye for 3D analyses.
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