Multi-scale alignment of respiratory cilia and its relation to mucociliary function.

The tracheobronchial tree is lined by a mucociliary epithelium containing millions of multiciliated cells. Their integrated oscillatory activity continuously propels an overlying pollution-protecting mucus layer in cranial direction, leading to mucociliary clearance - the primary defence mechanism of the airways. Mucociliary transport is commonly thought to co-emerge with the collective ciliary motion pattern under appropriate geometrical and rheological conditions. Proper ciliary alignment is therefore considered essential to establish mucociliary clearance in the respiratory system. Here, we used volume electron microscopy in combination with high-speed reflection contrast microscopy in order to examine ciliary orientation and its spatial organization, as well as to measure the propagation direction of metachronal waves and the direction of mucociliary transport on bovine tracheal epithelia with reference to the tracheal long axis (TLA). Ciliary orientation is measured in terms of the basal body orientation (BBO) and the axonemal orientation (AO), which are commonly considered to coincide, both equivalently indicating the effective stroke as well as the mucociliary transport direction. Our results, however, reveal that only the AO is in line with the mucociliary transport, which was found to run along a left-handed helical trajectory, whereas the BBO was found to be aligned with the TLA. Furthermore, we show that even if ciliary orientation remains consistent between adjacent cells, ciliary orientation exhibits a gradual shift within individual cells. Together with the symplectic beating geometry, this intracellular orientational pattern could provide for the propulsion of highly viscous mucus and likely constitutes a compromise between efficiency and robustness.

Main airway throughout the nasal cavity of green sea turtles is lined by keratinized stratified squamous epithelium.

Structural and histological features of the nasal cavity of sea turtles are largely different from those of other Testudines species. The sea turtle nasal cavity is a pair of tubular structures with three diverticula and an excavation in the center, and three types of sensory epithelium are present in these four significant structures. To more clarify the adaptation of the nasal cavity to marine life style in sea turtles, non-sensory epithelium in the nasal cavity of green sea turtles (Chelonia mydas) were histologically determined from nostril to choanae in this study. Unlike many other animals including terrestrial turtles, the vestibular area and nasopharyngeal duct were all lined by keratinized stratified squamous epithelium. In the main nasal cavity, the margins of each sensory epithelium turned into respiratory epithelium with goblet cells, followed by keratinized stratified squamous epithelium. Keratinized epithelium appears more appropriate in sea turtle upper airway to protect against osmotic pressure when they release seawater through the nostrils, and thus this histological feature of upper airway might reflect adaptation to marine life style.

On the functional compartmentalization of the normal middle ear. Morpho-histological modelling parameters of its mucosa.

BACKGROUND: Middle ear physiology includes both sound pressure transmission and homeostasis of its static air pressure. Pressure gradients are continuously created by gas exchange over the middle ear mucosa as well as by ambient pressure variations. Gas exchange models require actual values for regional mucosa thickness, blood vessel density, and diffusion distance. Such quantitative data have been scarce and limited to few histological samples from the tympanic cavity (TC) and the antrum. However, a detailed regional description of the morphological differences of the TC and mastoid air cell system (MACS) mucosa has not been available. The aim of the present study was to provide such parameters. METHODS: The study included sets of three histological H&E-slides from 15 archived healthy temporal bones. We performed a comparison of the mucosa morphology among the following regions: (1) anterior TC; (2) inferior TC; (3) posterior TC; (4) superior TC; (5) MACS antrum; (6) superior MACS; (7) central MACS; (8) inferior MACS. RESULTS: Regions (1)-(3), situated below the inter-attico-tympanic diaphragm, had the largest proportion of high respiratory epithelium, cilia and loose lamina propria within the mucosa, as well as the thickest mucosa and the largest diffusion distance. Regions (6)-(8), situated above the diaphragm, had the thinnest mucosa, the shortest distance to the blood vessels, together with the largest proportion of flat epithelium and very few cilia. Regions (4)-(5), still supradiaphragmatic, had intermediary values for these parameters, but generally closer to regions (6)-(8). The blood vessel density and the proportion of active mucosa were not significantly different among the regions. CONCLUSION: Mucosa of regions (1), (2) and (3) represented a predominantly clearance-specific morphology, whereas in regions (4)-(8) it seemed adapted to gas exchange. However, the lack of statistically significant differences in blood vessel density and proportion of active mucosa indicated that all regions could be involved in gas exchange with the highest adaptation in the superior MACS. This pattern divides the middle ear functionally along the inter-attico-tympanic diaphragm rather than the anatomical division into TC and MACS.

Comparative View of Lung Vascular Endothelium of Cattle, Horses, and Water Buffalo.

Endothelium plays an important role in maintaining the vascular barrier and physiological homeostasis. Endothelium also is fundamental to the initiation and regulation of inflammation. Endothelium demonstrates phenotypic and functional heterogeneity not only among various organs but also within an organ. One of the striking examples would be the pulmonary endothelium that participates in creating blood-air barrier. Endothelium in large pulmonary blood vessels is distinct in structure and function from that lining of the pulmonary capillaries. This chapter focuses on the comparative aspects of pulmonary endothelium and highlight unique differences such as the presence of pulmonary intravascular macrophages among select species.

Histological Characteristics of the Tracheobronchial Tree of the Least Shrew (Cryptotis Parva).

The least shrew (Cryptotis parva) is a small vomit-competent insectivorous species which has recently been introduced as an emesis animal model in the laboratory. In this study, the respiratory system of the least shrew was examined and compared with the well-established larger species routinely used in the laboratory. Five least shrews (4-5 g body weight, 45-60 days old) were used. Standard histological procedures were followed for light microscopic examination. The lining epithelium of the trachea was found to be pseudostratified ciliated columnar (PSCC). Three types of cells were easily identified, basal and ciliated as well as few goblet cells interspersed among the ciliated cells and they were not clearly recognizable. A few tracheal seromucous glands were located at the free end of the C-shaped cartilaginous rings. The cartilaginous rings are replaced by smooth muscle cells before the bronchi enter into the lung. The lining epithelium of tracheobronchial tree gradually changes into simple cuboidal epithelium that lacks goblet cells. However, the division of the tracheobronchial tree is similar to other mammalian species. On the other hand, the principal bronchus lacks cartilaginous plaques as it becomes intrapulmonary bronchus. The wall of the bronchi is supported by thick layers of spirally arranged smooth muscles. Two types of cells were readily recognizable: basal and ciliated cells, with rarely observed goblet cells. In addition, the PSCC epithelium changes into simple cuboidal much earlier in the bronchial division relative to other species.

Tolerance and pharmacokinetics of a ciprofloxacin-coated sinus stent in a preclinical model.

BACKGROUND: Chronic rhinosinusitis (CRS) is often associated with persistent bacterial infection despite the use of systemic antibiotics. Topically administered antibiotics are an alternative strategy, but require effective local concentrations, prolonged mucosal contact time, minor systemic absorption, and minimal depletion. The objectives of the current study were to analyze the in vitro release rate and in vivo drug delivery tolerance and pharmacokinetics of a ciprofloxacin-coated sinus stent (CSS). METHODS: The CSS (2 mg) was created from biodegradable poly-D/L-lactic acid. After analyzing in vitro release profile, CSSs were placed unilaterally in maxillary sinuses of 16 rabbits via dorsal sinusotomy. Animals were euthanized between 1 and 3 weeks postoperatively. Ciprofloxacin concentrations in the sinus tissue and plasmas were assessed using high-performance liquid chromatography. Radiological and histological evaluations were performed. RESULTS: In the in vitro release profile, an initial burst release was observed over the first 24 hours, followed by sustained release through the 14-day time point. In the rabbit model, ciprofloxacin was continuously released from the stent up to 3 weeks at doses >50 ng/mL. Histologic examination found no evidence of inflammation, epithelial ulceration, or bony reaction upon euthanization of the animals at 21 days. Computed tomography also demonstrated no signs of mucosal edema or opacification in the sinus. CONCLUSION: The CSS was safe in this preclinical model and sustained release was observed in both the in vitro and in vivo analyses. The innovative stent design coated with ciprofloxacin may provide a unique therapeutic strategy for chronic rhinosinusitis (CRS).

Epithelial Notch signaling regulates lung alveolar morphogenesis and airway epithelial integrity.

Abnormal enlargement of the alveolar spaces is a hallmark of conditions such as chronic obstructive pulmonary disease and bronchopulmonary dysplasia. Notch signaling is crucial for differentiation and regeneration and repair of the airway epithelium. However, how Notch influences the alveolar compartment and integrates this process with airway development remains little understood. Here we report a prominent role of Notch signaling in the epithelial-mesenchymal interactions that lead to alveolar formation in the developing lung. We found that alveolar type II cells are major sites of Notch2 activation and show by Notch2-specific epithelial deletion (Notch2(cNull)) a unique contribution of this receptor to alveologenesis. Epithelial Notch2 was required for type II cell induction of the PDGF-A ligand and subsequent paracrine activation of PDGF receptor-α signaling in alveolar myofibroblast progenitors. Moreover, Notch2 was crucial in maintaining the integrity of the epithelial and smooth muscle layers of the distal conducting airways. Our data suggest that epithelial Notch signaling regulates multiple aspects of postnatal development in the distal lung and may represent a potential target for intervention in pulmonary diseases.

Hajek revisited: a histological examination of the quadrangular membrane.

OBJECTIVE: The predictability of laryngeal cancer spread is due in part to connective tissue membranes. These membranes function as barriers to cancer and divide the larynx into subunits. The field of laryngeal conservation surgery is based on these concepts. The quadrangular membrane plays an important role, hindering the lateral spread of cancer in the larynx. The composition of this membrane has not been well described in the literature. In this study, we examine basic characteristics of the quadrangular membrane using histological techniques. METHODS: Whole organ sections of the larynx were used. These sections were examined under a microscope with stains specific for collagen and elastin. RESULTS: Examination of the sections revealed that the quadrangular membrane is made up of closely woven undulating collagen and elastic fibers. CONCLUSION: The quadrangular membrane is a fibroelastic structure providing a barrier to cancer spread.

Histological and lectin histochemical studies on the olfactory and respiratory mucosae of the sheep.

The olfactory and respiratory mucosae of the Corriedale sheep were examined using lectin histochemistry in order to clarify the histochemical and glycohistochemical differences between these two tissues. The olfactory epithelium was stained with 13 lectins out of 21 lectins examined, while the respiratory epithelium was positive to 16 lectins. The free border of both of the olfactory and respiratory epithelia was stained with 12 lectins: Wheat germ agglutinin (WGA), succinylated-wheat germ agglutinin (s-WGA), Lycopersicon esculentum lectin (LEL), Solanum tuberosum lectin (STL), Datura stramonium lectin (DSL), Soybean agglutinin (SBA), Bandeiraea simplicifolia lectin-I (BSL-I), Ricinus communis agglutinin-I (RCA-120), Erythrina cristagalli lectin (ECL), Concanavalin A (Con A), Phaseolus vulgaris agglutinin-E (PHA-E) and Phaseolus vulgaris agglutinin-L (PHA-L). The associated glands of the olfactory mucosa, Bowman's glands, were stained with 13 lectins. While both the goblet cells and mucous nasal glands were stained with 8 lectins; five of them (WGA, s-WGA, STL, Vicia villosa agglutinin (VVA) and ECL) were mutually positive among the Bowman's glands, mucous nasal glands and the goblet cells. These findings indicate that the glycohistochemical characteristics of the free borders of both olfactory and respiratory epithelia are similar to each other, suggesting that secretions from the Bowman's glands and those of the goblet cells and mucous nasal glands are partially exchanged between the surface of two epithelia to contribute the functions of the respiratory epithelium and the olfactory receptor cells, respectively.

Tonsil surface epithelium is ideal for monitoring Ki-67 immunohistochemical staining.

AIMS: To identify an easily obtainable non-neoplastic tissue that can be used as control material for monitoring optimal Ki-67 immunohistochemical staining. METHODS AND RESULTS: Various tissues, including tonsil (60), uterine cervix (31), breast skin (26), oesophagus (15), stomach (15), small intestine (15) and colon (16), were studied in the search for ideal control tissue. Tonsil surface epithelium is superior to other tissues because it displayed an easily recognized Ki-67 staining pattern including high positive (parabasal layer), low positive (intermediate layer) and negative (basal and superficial layers) zones. Moderate to weak staining of the majority of the intermediate cells could serve as a threshold for positive staining. Of the variables potentially affecting staining results that were tested, the pretreatment solution for antigen retrieval had the greatest impact, of which pH 9 EDTA was far better than pH 6 citrate solution. CONCLUSIONS: Tonsil surface epithelium is a useful control for monitoring Ki-67 staining. Achieving optimal staining results could minimize variations in Ki-67 index due to differences in the staining methods used by different laboratories.

Surface fluid absorption and secretion in small airways.

Native small airways must remain wet enough to be pliable and support ciliary clearance, but dry enough to remain patent for gas flow. The airway epithelial lining must both absorb and secrete ions to maintain a critical level of fluid on its surface. Despite frequent involvement in lung diseases, the minuscule size has limited studies of peripheral airways. To meet this challenge, we used a capillary to construct an Ussing chamber (area <1 mm(2)) to measure electrolyte transport across small native airways (∼1 mm ø) from pig lung. Transepithelial potentials (V(t)) were recorded in open circuit conditions while applying constant current pulses across the luminal surface of dissected airways to calculate transepithelial electrical conductance (G(t)) and equivalent short circuit current (I(eq)(sc)) in the presence and absence of selected Na(+) and Cl(-) transport inhibitors (amiloride, GlyH-101, Niflumic acid) and agonists (Forskolin + IBMX, UTP). Considered together the responses suggest an organ composed of both secreting and absorbing epithelia that constitutively and concurrently transport fluids into and out of the airway, i.e. in opposite directions. Since the epithelial lining of small airways is arranged in long, accordion-like rows of pleats and folds that run axially down the lumen, we surmise that cells within the pleats are mainly secretory while the cells of the folds are principally absorptive. This structural arrangement could provide local fluid transport from within the pleats toward the luminal folds that may autonomously regulate the local surface fluid volume for homeostasis while permitting acute responses to maintain clearance.

Microscopic anatomy of the lower respiratory tract of the grey short-tailed opossum (Monodelphis domestica).

The respiratory tracts of seven grey short-tailed opossums were histologically examined. Six opossums were prepared by perfusion with buffered formalin. Opossum seven was perfused with gluteraldehyde. Samples taken from the respiratory passages and lungs of specimens 1-6 were stained with haematoxylin and eosin. A mixture of methylene and azure blue was used for specimen 7. The trachea and right and left principal bronchi are lined with a pseudostratified ciliated columnar epithelium with occasional goblet cells. The secondary and tertiary bronchi and the primary and secondary bronchioles are lined by a simple ciliated columnar epithelium. The terminal bronchioles and a portion of the respiratory bronchioles are lined by a simple ciliated cuboidal epithelium. The terminal portion of the respiratory bronchioles and the alveolar ducts are lined with simple squamous epithelium. Alveoli are lined by type I and II pneumocytes. Tracheal glands are present in the tela submucosa. The fibromusculocartilaginous tunic of the trachea consists of c-shaped cartilage rings and the trachealis muscle. A lamina muscularis mucosa begins in the intrapulmonary portion of the principal bronchus and continues into the respiratory bronchioles. Bronchial glands are present in the propria submucosa and tela submucosa of the principal bronchi. The musculocartilaginous tunic is localized to the extrapulmonary portion of the principal bronchus. The bronchial cartilages are irregular shaped plates and limited to the extrapulmonary portion of the principal bronchus. The visceral pleura is a simple squamous mesothelium covering the outer surface of the lung.

Human cystic fibrosis airway epithelia have reduced Cl- conductance but not increased Na+ conductance.

Loss of cystic fibrosis transmembrane conductance regulator (CFTR) anion channel function causes cystic fibrosis (CF) lung disease. CFTR is expressed in airway epithelia, but how CF alters electrolyte transport across airway epithelia has remained uncertain. Recent studies of a porcine model showed that in vivo, excised, and cultured CFTR(-/-) and CFTR(ΔF508/ΔF508) airway epithelia lacked anion conductance, and they did not hyperabsorb Na(+). Therefore, we asked whether Cl(-) and Na(+) conductances were altered in human CF airway epithelia. We studied differentiated primary cultures of tracheal/bronchial epithelia and found that transepithelial conductance (Gt) under basal conditions and the cAMP-stimulated increase in Gt were markedly attenuated in CF epithelia compared with non-CF epithelia. These data reflect loss of the CFTR anion conductance. In CF and non-CF epithelia, the Na(+) channel inhibitor amiloride produced similar reductions in Gt and Na(+) absorption, indicating that Na(+) conductance in CF epithelia did not exceed that in non-CF epithelia. Consistent with previous reports, adding amiloride caused greater reductions in transepithelial voltage and short-circuit current in CF epithelia than in non-CF epithelia; these changes are attributed to loss of a Cl(-) conductance. These results indicate that Na(+) conductance was not increased in these cultured CF tracheal/bronchial epithelia and point to loss of anion transport as key to airway epithelial dysfunction in CF.

A lumped mucosal wave model of the vocal folds revisited: recent extensions and oscillation hysteresis.

This paper examines an updated version of a lumped mucosal wave model of the vocal fold oscillation during phonation. Threshold values of the subglottal pressure and the mean (DC) glottal airflow for the oscillation onset are determined. Depending on the nonlinear characteristics of the model, an oscillation hysteresis phenomenon may occur, with different values for the oscillation onset and offset threshold. The threshold values depend on the oscillation frequency, but the occurrence of the hysteresis is independent of it. The results are tested against pressure data collected from a mechanical replica of the vocal folds, and oral airflow data collected from speakers producing intervocalic /h/. In the human speech data, observed differences between voice onset and offset may be attributed to variations in voice pitch, with a very small or inexistent hysteresis phenomenon.

Microscopic anatomy of the lower respiratory system of the African giant pouched rat (Cricetomys gambianus, Waterhouse 1840) / Anatomía microscópica del sistema respiratorio inferior de la rata gigante Africana (Cricetomys gambianus, Waterhouse 1840)

A qualitative and quantitative study, by light microscopy, was undertaken on the lower respiratory system of the African Giant pouched rat. Specifically, the trachea, bronchi and lungs were stained with Haematoxylin and eosin, Alcian blue at a pH of 2.5 and Periodic Acid-Schiff stains. Three cell types were identified in saggital sections of the trachea: the ciliated cells, basal cells and mucous cells. Fibers of the trachealis muscles in the laminar propria separated the underlying cartilages from the basal cells. Mucous cells were visible only in the membranous portion of the trachea and they were predominant in the rostral and caudal portion of the trachea. Lobar bronchi consisted of cuboidal epithelium and a layer of one or two smooth muscle cells and opened into segmental bronchi and respiratory bronchiole. Some tracheal cartilaginous rims stained blue with AB while most glandular cells stained red with PAS. The diameter of respiratory bronchiole, alveoli duct and alveoli were 24.93 µm (+/- 1.27), 21.14 um (+/- 0.66) and 12.95 um (+/- 0.21), respectively. These and other findings were compared with similar report in other rodents.

Se realizó un estudio cualitativo y cuantitativo, mediante microscopía de luz, en el sistema respiratorio inferior de la rata gigante Africana. La tráquea, los bronquios y los pulmones fueron teñidos con hematoxilina y eosina, azul Alcián a pH de 2,5 y ácido periódico de Schiff. Tres tipos de células fueron identificadas en las secciones sagitales de la tráquea: células ciliadas, basales y mucosas. Las fibras del músculo traqueal en la propia laminar separados los cartílagos subyacente de las células basales. las células mucosas son visibles sólo en la porción membranosa de la tráquea y predominan en la parte rostral de la porción caudal de la tráquea. Los bronquios lobares consistían en epitelio cúbico y una capa de una o dos células de músculo liso y abierto en los bronquios y bronquiolos segmentarios respiratorias. Algunos bordes azules cartilaginoso traqueal manchada con AB, mientras que la mayoría de las células glandulares teñido de rojo con PAS. El diámetro de los bronquiolos respiratorios, conductos alveolares y los alvéolos fueron 24,93 m (+/- 1,27), 21,14 m (+/- 0,66) y 12,95 m (+/- 0,21), respectivamente. Estos y otros resultados se compararon con el informe similar en otros roedores.

Development of the bronchial epithelial reticular basement membrane: relationship to epithelial height and age.

BACKGROUND: The bronchial epithelium and underlying reticular basement membrane (RBM) have a close spatial and functional inter-relationship and are considered an epithelial-mesenchymal trophic unit (EMTU). An understanding of RBM development is critical to understanding the extent and time of appearance of its abnormal thickening that is characteristic of asthma. METHODS: RBM thickness and epithelial height were determined in histological sections of cartilaginous bronchi obtained postmortem from 47 preterm babies and infants (median age 40 weeks gestation (22 weeks gestation-8 months)), 40 children (2 years (1 month-17 years)) and 23 adults (44 (17-90) years) who had died from non-respiratory causes, and had no history of asthma. RESULTS: The RBM was visible by light microscopy at 30 weeks gestation. RBM thickness increased in successive age groups in childhood; in infants (r=0.63, p<0.001) and in children between 1 month and 17 years (r=0.82, p<0.001). After 18 years, RBM thickness decreased with increasing age (r=-0.42, p<0.05). Epithelial height showed a similar relationship with age, a positive relationship from preterm to 17 years (r=0.50, p<0.001) and a negative relationship in adulthood (r=-0.84, p<0.0001). There was a direct relationship between epithelial height and RBM thickness (r=0.6, p<0.001). CONCLUSIONS: The RBM in these subjects was microscopically identifiable by 30 weeks gestation. It thickened during childhood and adolescence. In adults, there was either no relationship with age, or a slow reduction in thickness in older age. Developmental changes of RBM thickness were accompanied by similar changes in epithelial height, supporting the close relationship between RBM and epithelium within the EMTU.

[Observations on the anatomy and physiology of the mammalian noses]. / Betrachtungen zur Anatomie und Physiologie von Säugetiernasen.

From the anatomical and flow dynamic point of view, the mammalian noses are strictly divided in a respiratory and olfactory area. In humans, the middle and the superior turbinate (ethmoturbinates) fulfill the respiratory and olfactory function. An intensive contact between breathing air and respiratory mucosa is the most important pre-condition for the respiratory function of the nose. In all investigated species this contact takes place in the functional area of the nose, i. e. the area of the turbinates. In humans, the area from the external nasal ostium up to the head of the inferior turbinate is called "inflow area". This part of the nose distributes the airflow over the complete nasal cross sectional area and generates turbulent flow. The inflow area in the human nose is an adaptation to the relatively increased nasal height during evolution.

Postnatal development of the lamina reticularis in primate airways.

The basement membrane zone (BMZ) appears as three component layers: the lamina lucida, lamina densa, and lamina reticularis. The laminas lucida and densa are present during all stages of development. The lamina reticularis appears during postnatal development. Collagens I, III, and V form heterogeneous fibers that account for the thickness of the lamina reticularis. Additionally, there are three proteoglycans considered as integral components of the BMZ: perlecan, collagen XVIII, and bamacan. Perlecan is the predominant heparan sulfate proteoglycan in the airway BMZ. It is responsible for many of the functions attributed to the BMZ, in particular, trafficking of growth factors and cytokines between epithelial and mesenchymal cells. Growth factor binding sites on perlecan include FGF-1, FGF-2, FGF-7, FGF-10, PDGF, HGF, HB-EGF, VEGF, and TGF-beta. Growth factors pass through the BMZ when moving between the epithelial and mesenchymal cell layers. They move by rapid reversible binding with sites on both the heparan sulfate chains and core protein of perlecan. In this manner, perlecan regulates movement of growth factors between tissues. Another function of the BMZ is storage and regulation of FGF-2. FGF-2 has been shown to be involved with normal growth and thickening of the BMZ. Thickening of the BMZ is a feature of airway remodeling in asthma. It may have a positive effect by protecting against airway narrowing and air trapping. Conversely, it may have a negative effect by influencing trafficking of growth factors in the epithelial mesenchymal trophic unit. However, currently the significance of BMZ thickening is not known.

Heparan sulfates in the lung: structure, diversity, and role in pulmonary emphysema.

There is an emerging interest in the extracellular matrix (ECM) of the lung, especially in the role it plays in development and disease. There is a rapid change from the classical view of the ECM as a supporting structure towards a view of the ECM as a regulatory entity with profound effects on proliferation, migration, and differentiation of pulmonary cells. In the ECM, a variety of molecules is present in a highly organized pattern. Next to the abundant fiber-forming molecules such as collagens and elastin, a large number of less abundant molecules are part of the ECM, including proteoglycans. In this review, we will focus on one class of proteoglycans, the heparan sulfate proteoglycans. We will particularly address the structure, biosynthesis, and function of their saccharide moiety, the heparan sulfates, including their role in development and (patho)physiology.

Laser scanning microscopy combined with image restoration to analyse a 3D model of the human epithelial airway barrier.

A laser scanning microscope collects information from a thin, focal plane and ignores out of focus information. During the past few years it has become the standard imaging method to characterise cellular morphology and structures in static as well as in living samples. Laser scanning microscopy combined with digital image restoration is an excellent tool for analysing the cellular cytoarchitecture, expression of specific proteins and interactions of various cell types, thus defining valid criteria for the optimisation of cell culture models. We have used this tool to establish and evaluate a three dimensional model of the human epithelial airway wall.