Alterations of mouse lung tissue dimensions during processing for morphometry: A comparison of methods

Schneider, Jan Philipp; Ochs, Matthias

doi:10.1152/ajplung.00329.2013

Cited by 85 publications

(69 citation statements)

References 46 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The unstained uterus sample decreased in volume by 44.8%, while the stained uterus sample decreased by 19.2%. On the other hand, the unstained lung sample decreased in volume by 60.5% while the stained lung sample decreased in volume by 57.3%, which is consistent with the results presented previously [12]. This difference in volume change between the two tissue types is surprising and may be partially attributed to variations in the machine resolution for different scan parameters, segmentation errors, differences in tissue sample density and difficulty imaging the embedded, unstained lung tissue with micro-CT. Pre-embedded images of the unstained lung sample were acquired without problem, but the embedded sample poorly attenuated micro-CT X-rays, which made segmentation and volume measurements difficult and imprecise.…”

Section: Discussionsupporting

confidence: 93%

“…As a result, the lung tissue would have been more likely to lose more fluid during the embedding process than either the uterus or mouse brain tissue, leading to the greatest decrease in tissue volume among the three samples. Such variation in tissue shrinkage could impact the accuracy of quantitative measurements in tissue [12]. Information on the shrinkage characteristics of the different tissue types that we initially gathered in this work gives valuable inputs for designing an experimental setup involving a larger number of samples to come up with an effective embedding protocol for each tissue type, such as using wax for embedding [11,13].…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

The Role of Micro-CT in 3D Histology Imaging

et al. 2016

View full text Add to dashboard Cite

Objectives: 3D histology tissue modeling is a useful analytical technique for understanding anatomy and disease at the cellular level. However, the current accuracy of 3D histology technology is largely unknown, and errors, misalignment and missing information are common in 3D tissue reconstruction. We used micro-CT imaging technology to better understand these issues and the relationship between fresh tissue and its 3D histology counterpart. Methods: We imaged formalin-fixed and 2% Lugol-stained mouse brain, human uterus and human lung tissue with micro-CT. We then conducted image analyses on the tissues before and after paraffin embedding using 3D Slicer and ImageJ software to understand how tissue changes between the fixation and embedding steps. Results: We found that all tissue samples decreased in volume by 19.2-61.5% after embedding, that micro-CT imaging can be used to assess the integrity of tissue blocks, and that micro-CT analysis can help to design an optimized tissue-sectioning protocol. Conclusions: Micro-CT reference data help to identify where and to what extent tissue was lost or damaged during slide production, provides valuable anatomical information for reconstructing missing parts of a 3D tissue model, and aids in correcting reconstruction errors when fitting the image information in vivo and ex vivo.

show abstract

Section: Discussionsupporting

confidence: 93%

Section: Discussionmentioning

confidence: 99%

The Role of Micro-CT in 3D Histology Imaging

et al. 2016

View full text Add to dashboard Cite

show abstract

“…This indicates lagged alveolar development, which matches the pathological characteristics of a new BPD displaying a pulmonary developmental disorder. Meanwhile, we must realize that there are pitfalls of the morphometric analysis using paraffin-embedded tissue because tissue-processing procedures may cause nearly 40% shrinkage of the lung tissue (48).…”

Section: Discussionmentioning

confidence: 99%

Hyperoxia stimulates the transdifferentiation of type II alveolar epithelial cells in newborn rats

Hou

Yang

et al. 2015

American Journal of Physiology-Lung Cellular and Molecular Phys

View full text Add to dashboard Cite

-Supplemental oxygen treatment in preterm infants may cause bronchopulmonary dysplasia (BPD), which is characterized by alveolar simplification and vascular disorganization. Despite type II alveolar epithelial cell (AEC II) damage being reported previously, we found no decrease in the AEC II-specific marker, surfactant protein C (SP-C), in the BPD model in our previous study. We thus speculated that AEC II injury is not a unique mechanism of BPD-related pulmonary epithelial repair dysfunction and that abnormal transdifferentiation can exist. Newborn rats were randomly assigned to model (85% oxygen inhalation) and control groups (room air inhalation). Expressions of AEC I (aquaporin 5, T1␣) and AEC II markers (SP-C, SP-B) were detected at three levels: 1) in intact lung tissue, 2) in AEC II isolated from rats in the two groups, and 3) in AEC II isolated from newborn rats, which were further cultured under either hyperoxic or normoxic conditions. In the model group, increased AEC I was observed at both the tissue and cell level, and markedly increased transdifferentiation was observed by immunofluorescent double staining. Transmission electron microscopy revealed morphological changes in alveolar epithelium such as damaged AECs, a fused air-blood barrier structure, and opened tight junctions in the model group. These findings indicate that transdifferentiation of AECs is not suppressed but rather is increased under hyperoxic treatment by compensation; however, such repair during injury cannot offset pulmonary epithelial air exchange and barrier dysfunction caused by structural damage to AECs. transdifferentiation; alveolar epithelial cells; hyperoxia; bronchopulmonary dysplasia SUPPLEMENTAL OXYGEN TREATMENT, which is often necessary for preterm infants with respiratory failure, has become a major risk factor for bronchopulmonary dysplasia (BPD), one of the most common, serious complications in preterm infants (6,34). BPD is characterized by high morbidity in infants with very low birth weight (BW Ͻ 1,500 g) soon after birth and may induce multiple complications in respiratory and nervous systems during adolescence (4, 6). Despite several decades of research, the underlying pathophysiological foundation of BPD has not been completely clarified.Lung tissue consists of many different cell types, and a developmental disorder of alveolar epithelial cells (AEC) is likely a major cause of BPD (40). The alveolar area accounts for Ն99% of the internal surface area of the lungs (49). Mammals have two types of AECs that maintain normal air exchange function by mutual coordination. Type I AEC (AEC I) cover nearly 96% of alveolar spaces and closely adhere to adjacent capillaries. Such cells are the main epithelial constituents of the air-blood barrier (18, 53) and have air exchange functions. Recent studies have revealed that AEC I can also secrete transport proteins to maintain intrapulmonary fluid and electrolyte balance (9,19,22). Type II AEC (AEC II) are located at the corners of alveoli and are more abundant than AEC I bu...

show abstract

“…In such instances, minor alterations to lung parenchymal structure may not be detected, or may be exaggerated, being either magnified or masked. To avoid this, a state-of-the-art analysis of the lung structure was undertaken (65). This analysis includes the preservation of lung structure, followed by SUR sampling, and a stereological analysis allowing for the determination of the total number of alveoli, septal thickness, and MLI (66).…”

Section: Discussionmentioning

confidence: 99%

Systemic hydrogen sulfide administration partially restores normal alveolarization in an experimental animal model of bronchopulmonary dysplasia

Madurga

Mižíková

Ruiz‐Camp

et al. 2014

American Journal of Physiology-Lung Cellular and Molecular Phys

View full text Add to dashboard Cite

Arrested alveolarization is the pathological hallmark of bronchopulmonary dysplasia (BPD), a complication of premature birth. Here, the impact of systemic application of hydrogen sulfide (H2S) on postnatal alveolarization was assessed in a mouse BPD model. Exposure of newborn mice to 85% O2 for 10 days reduced the total lung alveoli number by 56% and increased alveolar septal wall thickness by 29%, as assessed by state-of-the-art stereological analysis. Systemic application of H2S via the slow-release H2S donor GYY4137 for 10 days resulted in pronounced improvement in lung alveolarization in pups breathing 85% O2, compared with vehicle-treated littermates. Although without impact on lung oxidative status, systemic H2S blunted leukocyte infiltration into alveolar air spaces provoked by hyperoxia, and restored normal lung interleukin 10 levels that were otherwise depressed by 85% O2. Treatment of primary mouse alveolar type II (ATII) cells with the rapid-release H2S donor NaHS had no impact on cell viability; however, NaHS promoted ATII cell migration. Although exposure of ATII cells to 85% O2 caused dramatic changes in mRNA expression, exposure to either GYY4137 or NaHS had no impact on ATII cell mRNA expression, as assessed by microarray, suggesting that the effects observed were independent of changes in gene expression. The impact of NaHS on ATII cell migration was attenuated by glibenclamide, implicating ion channels, and was accompanied by activation of Akt, hinting at two possible mechanisms of H2S action. These data support further investigation of H2S as a candidate interventional strategy to limit the arrested alveolarization associated with BPD.

show abstract

Alterations of mouse lung tissue dimensions during processing for morphometry: A comparison of methods

Cited by 85 publications

References 46 publications

The Role of Micro-CT in 3D Histology Imaging

The Role of Micro-CT in 3D Histology Imaging

Hyperoxia stimulates the transdifferentiation of type II alveolar epithelial cells in newborn rats

Systemic hydrogen sulfide administration partially restores normal alveolarization in an experimental animal model of bronchopulmonary dysplasia

Contact Info

Product

Resources

About