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![Cilia a and b. Solitary peripheral tubules [arrows on both sides of the cilium with double tubules between them (thick arrows)]. Dynein arms cannot be seen in these double tubules of transformation zone. Cilia c and d .](profile/Markus-Rautiainen/publication/16711430/figure/fig2/AS:525328879964160@1502259573510/FIG-7-Cilia-a-and-b-Solitary-peripheral-tubules-arrows-on-both-sides-of-the-cilium.png)
Cilia a and b. Solitary peripheral tubules [arrows on both sides of the cilium with double tubules between them (thick arrows)]. Dynein arms cannot be seen in these double tubules of transformation zone. Cilia c and d .
Source publication
Ultrastructure of human respiratory cilia was studied in ultrathin serial sections and in random fields of sections. The apex of the cilium had dense gray material under the plasma membrane, where the "central" pair of tubules (often slightly eccentrically placed) and the single peripheral tubules ended. Thereafter the cilium thickened gradually ab...
Citations
... This geometry changes through the axoneme, reducing the doublets angulation but keeping the rotational symmetry of the basal body [15]. Consequently, peripheral doublets show a slight non-pathologic tilt, evidenced in 1984 by Rautiainen et al. in a study of ciliary serial sections [16]. We have termed this conformation as "axonemal symmetry". ...
Diagnosis testing for primary ciliary dyskinesia (PCD) requires a combination of investigations that includes study of ciliary beat pattern by high-speed video-microscopy, genetic testing and assessment of the ciliary ultrastructure by transmission electron microscopy (TEM). Historically, TEM was considered to be the “gold standard” for the diagnosis of PCD. However, with the advances in molecular genetic techniques, an increasing number of PCD variants show normal ultrastructure and cannot be diagnosed by TEM. During ultrastructural assessment of ciliary biopsies of patients with suspicion of PCD, we observed an axonemal defect not previously described that affects peripheral doublets tilting. To further characterize this defect of unknown significance, we studied the ciliary axonemes by TEM from both PCD-confirmed patients and patients with other sino-pulmonary diseases. We detected peripheral doublets tilting in all the PCD patients, without any significant difference in the distribution of ciliary beat pattern or mutated gene. This defect was also present in those patients with normal ultrastructure PCD subtypes. We believe that the performance of axonemal asymmetry analysis would be helpful to enhance diagnosis of PCD.
... These may interfere with subsequent alignment of the dataset. Even worse than artifacts is that entire sections may become useless, for example, because of folds or film damage (Young et al. 1985) or may be lost completely (Rautiainen et al. 1984;see Saalfeld et al. 2012). The latter may result in the loss of important information necessary for correct structural interpretation or it may lead to a wrong "height" of the final model, if the loss is not corrected. ...
Since its entry into biomedical research in the first half of the twentieth century, electron microscopy has been a valuable tool for lung researchers to explore the lung's delicate ultrastructure. Among others, it proved the existence of a continuous alveolar epithelium and demonstrated the surfactant lining layer. With the establishment of serial sectioning transmission electron microscopy, as the first "volume electron microscopic" technique, electron microscopy entered the third dimension and investigations of the lung's three-dimensional ultrastructure became possible. Over the years, further techniques, ranging from electron tomography over serial block-face and focused ion beam scanning electron microscopy to array tomography became available. All techniques cover different volumes and resolutions, and, thus, different scientific questions. This review gives an overview of these techniques and their application in lung research, focusing on their fields of application and practical implementation. Furthermore, an introduction is given how the output raw data are processed and the final three-dimensional models can be generated.
... Each respiratory epithelial cell is lined with between 50 and 200 cilia, 1 which are about 5 μm to 6 μm long and about 0.2 μm wide. 2,3 The beating cilia are the driving force of mucociliary clearance, which is a process by which the mucus blanket overlying respiratory mucosa is transported to the gastrointestinal track for ingestion. Furthermore, mucociliary clearance is the primary mechanism by which the airway clears pathogens, allergens, debris, and toxins. 1 Various factors can affect cilia and mucociliary function. ...
Objectives/Hypothesis
The aim of this study was to evaluate statistically the effects of radiofrequency ablation, diode laser, and microdebrider‐assisted inferior turbinoplasty techniques on ciliated epithelium and mucociliary function.
Study Design
Prospective randomized study.
Methods
A total of 66 consecutively randomized adult patients with enlarged inferior turbinates underwent either a radiofrequency ablation, diode laser, or microdebrider‐assisted inferior turbinoplasty procedure. Assessments were conducted prior to surgery and 3 months subsequent to the surgery. The effect on ciliated epithelium was evaluated using a score based on the blinded grading of the preoperative and postoperative scanning electron microscopy images of mucosal samples. The effect on mucociliary function, in turn, was evaluated using saccharin transit time measurement.
Results
The score of the number of cilia increased statistically significantly in the radiofrequency ablation (P = .03) and microdebrider‐assisted inferior turbinoplasty (P = .04) groups, but not in the diode laser group. The score of the squamous metaplasia increased statistically significantly in the diode laser group (P = .002), but not in the other two groups. There were no significant changes found between the preoperative and postoperative saccharin transit time values in any of the treatment groups.
Conclusions
Radiofrequency ablation and microdebrider‐assisted inferior turbinoplasty are more mucosal preserving techniques than the diode laser, which was found to increase the amount of squamous metaplasia at the 3‐month follow‐up. The number of cilia seemed to even increase after radiofrequency ablation and microdebrider‐assisted inferior turbinoplasty procedures, but not after diode laser. Nevertheless, the mucociliary transport was equally preserved in all three groups.
Level of Evidence
1b. Laryngoscope, 2018
... In 2 smears, 1 mL of fluid was spun down, and the cell deposit was fixed in glutaraldehyde and examined by electron microscopy (EM) using standard protocol. 15 Patients were observed for 1-2 hours in the endoscopy recovery area, where careful monitoring of vital signs took place. An experienced nurse called patients routinely 24 -48 hours and at 30-day intervals to assess for potential complications more specifically inquiring about the development of fever, chest pain, or other symptoms perceived that have been related to the procedure. ...
... Transverse sections of the cilia showed the typical 9 ϩ 2 microtubule configuration (data not shown). 15 Multiple mitochondria were identified near the base of the cilia (Fig. 5). ...
The management of foregut duplication cysts is controversial, especially in asymptomatic patients. The safety and accuracy of endoscopic ultrasound (EUS) and EUS-fine-needle aspiration EUS-FNA) in confirming the nature of cysts by using electron microscopy (EM) has not been reported. In this study, the authors describe the utility of demonstrating detached ciliary tufts (DCTs) in the diagnosis of foregut duplication cysts with EUS-FNA.
Consecutive patients with suspected mediastinal masses or mediastinal cysts on imaging studies were evaluated prospectively by EUS and EUS-FNA. Cyst fluid was examined by routine cytologic techniques. In two patients, EM was performed to confirm the nature of DCTs.
Ten consecutive patients were evaluated with EUS and EUS-FNA. Seventy percent of the cysts were characterized by computed tomography (CT) scans as solid masses. The mean greatest cyst dimension measured 34 mm x 48 mm by EUS. Microscopic examination of the cyst content revealed mucinous material, cellular debris, and DCTs. The latter were seen in routine cytologic preparations and by EM. Patients were followed up to a median of 321 days. Due to EUS-FNA confirmatory diagnoses of foregut duplication cysts, none of the patients except 1 underwent surgical resection after developing pneumonia 6 months later. Histologic sections of the resected specimen confirmed the presence of (foregut cyst, bronchogenic type). All other patients were asymptomatic. Cysts size and nature did not change on repeated imaging studies.
EUS was superior compared with CT scanning in characterizing foregut duplication cysts. EUS-FNA is safe and accurate in the diagnosis of foregut duplication cysts. The demonstration of DCTs in cyst fluid and the absence of malignant cells confirmed the benign nature of these lesions, allowing conservative and expectant management for these patients.
... Approximately 0.50 pn below the apex peripheral microtubules begin to exist as doublets. This change occurs at what is referred to as the transfomation zone ( Rautiainen et al., 1984). Images of some cilia seen in Fig. 3 in the present study were sectioned at the transformation zone since a doublet configuration of microtubules was depicted in the axoneme profiles. ...
Oligociliated epithelial cells or cells with a solitary cilium were observed lining hepatic bile ductules in pigs. The cilia were situated in the midst of microvilli, and arose from a basal body that was located in the vicinity of the free surface of the epithelium. The axonemal pattern revealed in transverse sections of the cilia was 7 + 1, 8 + 1, 8 + 2 or 9 + 0, Although devoid of the classical axoneme pattern, we assume some of the cilia to be motile, and other to be sensory structures that would assist bile formation and secretion.
... Three additional biopsies were performed over 4-and 12-month intervals. The specimens were processed for electron microscopy by methods previously described [21,23]. Two of the samples were cut longitudinally perpendicular to the plane of the mucosa and examined in a JEOL JEM 100 S electron microscope. ...
... One piece of the second sample was serially cut longitudinally and used to measure the length of the cilia. The tips of the cilia were recognized as dark hazy dots (Fig. 2) [21]. ...
... The mean ciliary length (+ SD) measured in 19 longitudinal sections with whole cilia visible from the cell membrane to the tip was 2.5 + 0.9 ~tm (Fig. 4). The diameter of the ciliary shaft in the cross-sections was about 0.20 gm and was considered to be normal [21]. Occasionally, the cilia were very short and were only about 1 gm long. ...
A 48-year-old man was examined because of anosmia. He was otherwise healthy except for mild arterial hypertension. He had stopped smoking 20 years ago. Previously he had been exposed to sulfuric acid gases for 3 years in his work. Clinical examination revealed no findings to explain his anosmia. The patient was fertile, indicating normal sperm/cilia motility. Nasal mucociliary function was examined by radioactive tracer and found to be markedly and constantly impaired. Ciliary ultrastructure in cross-sections was normal. However, in longitudinal sections the length of the cilia varied from 0.6 microns to 3.9 microns. The mean length of the cilia from the cell membrane to the tip was 2.5 +/- 0.9 microns, in contrast to normal ciliary length of 5-7 microns. These findings represent a new structural defect among the various known ciliary abnormalities.
... In healthy human respiratory mucosa the alignment of the cilia may appear locally random, but larger populations of cilia result in more oriented patterns [22]. By estimating the ciliary orientation with the standard deviation of beat directions (as measured with a semiautomatic image analyzer), large populations of cilia can be covered [23]. In this study, we have measured ciliary orientation in patients with proven immotile cilia syndrome and have compared our findings with the ciliary orientation in control samples from healthy non-smokers. ...
... We also separated the measured fields into two groups according to the level of sectioning of the cilia: fields sectioned near the tip of the cilia or fields sectioned near the cell body. The presence of sectioned tips of cilia in the micrograph placed the field in the former group [23], the presence of microvilli in the latter. We also measured the length of the outer dynein arms in all patients and compared groups with different lengths of the dynein arms with each other. ...
Ciliary orientation was studied in 43 patients with the "immotile cilia" syndrome. Twenty-four of these patients had total situs inversus. One mucosal specimen was taken from uterine cervical epithelium, 2 were from bronchial mucosa and 40 from nasal mucosa. The orientation of the cilia was measured from micrographs using a semiautomatic image analyzer (IBAS I). The results from patients were compared with those of 10 control subjects. The mean standard deviation and its standard deviation of the angles of ciliary orientation was 39.7 degrees +/- 9.2 degrees in 43 patients and 27.4 degrees +/- 4.3 degrees in the control group. The difference between the groups is highly significant statistically (P less than or equal to 0.001). However, there were no statistically significant differences in the standard deviations of ciliary orientation between the fields sectioned near the cell membrane or near the ciliary tip. We were also unable to find any significant differences in the standard deviations of the ciliary angles in the specimens taken from brush biopsies and excisional biopsies. There were also no statistically significant differences between the standard deviations of the ciliary angles for the groups with or without situs inversus. If 35 degrees is considered to be the limit value for the mean standard deviation between normal and pathological specimens in our total material, this would give a specificity of 0.90 and a sensitivity of 0.72.
... These two parts of the motion cannot be distinguished from each other on the basis of ultrastructure of the crosssectioned cilia. The orientation can be measured on every level of cilia because cilia do not coil around their axis [21 ]. The measured angles are between 0-180°, because distinction could not be made between direction of stroke and recovery stroke. ...
Ciliary orientation was studied on the respiratory epithelium of the nasal cavity or the sphenoidal sinus of ten adult nonsmokers without respiratory disease. The ciliary orientation was evaluated from micrographs by measuring the angle between the plane defined by the central tubules and reference line (with a semiautomatic image analyser (IBAS I]. The standard deviation of the angles of cilia population was counted in every field. The standard deviation of the measurements described the ciliary alignment. It varied from 12.1-41.2 degrees. The mean standard deviation was 27.3 +/- 7.4 degrees. 58% of all measured cilia were within +/- 0-20 degrees of the mean and 85% of cilia were within +/- degrees. However, a few cilia or small groups of cilia were found in most fields which differed dramatically from the main orientation. The size of these groups was always less than ten cilia. On the normal respiratory epithelium the standard deviation of ciliary orientation varies between +/- 10-40 degrees (at about 97% probability). For diagnostic conclusions more than 60 cilia should be measured.
... Measurements of ciliary orientation can be done on any level of the cilia from the basal body to the tip of the cilia because the central tubules do not coil around the axis of each cilium [19]. Nonetheless the measurements of ciliary orientation must still be limited within 0~ ~ The reference line must be chosen so that the majority of the angles and the maximum point of their distribution are situated about the middle of the range. ...
We have created a method for measuring the orientation of cilia in the respiratory epithelium. Ciliary orientation is the direction perpendicular to the plane defined by the central tubules of the cilia and is an estimate of ciliary beat direction. Ciliary orientation can be estimated by measuring the angle between the plane defined by the central tubules and a reference line. The standard deviation of these measurements describes the variation present in the beat directions of the cilia. The reference line must be so chosen that the majority of measurements falls at about the middle of the 0 degree-180 degrees range. We tested measurements by using both a glass angle measure and a semiautomatic image analyzer (IBAS I). The latter approach was faster and more reproducible. We made our measurements of normal tissues on samples obtained from two healthy adult non-smokers. Measurements were made in four areas of each sample, with 59-110 cilia in each. The differences between the maximum and minimum angles of the ciliary orientation in the same area varied from 167.9 degrees to 85.4 degrees from the reference line. The standard deviation varied from 18.0 degrees to 35.4 degrees and we consider this to be a normal variation in ciliary orientation. Of the cilia, 57% were within a standard deviation of 20 degrees.
