Automated Whole Slide Morphometry (AWSM-Q): A Validation Study for Lung Histology Quantification in Rodent Models of Bronchopulmonary Dysplasia

Lindenmaier, Andras; Sadanand, Siddharth; Zheng, Jingjing; Zanette, Brandon; Friedlander, Yonni; Couch, Marcus J.; Stirrat, Elaine; Bouch, Sheena; Lok, Irene; Ivanovska, Julijana; Post, Martin; Jankov, Robert P.; Santyr, G.

doi:10.1164/ajrccm-conference.2019.199.1_meetingabstracts.a5492

Cited by 2 publications

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“…A composite score of lung injury was taken as the average of all scores for each caption and 8 captions were averaged for each animal. Automated whole slide morphometry was conducted based on a previously described protocol [ 14 , 15 ]. Digitized slides were automatically partitioned into 498 × 498 μm 2 tiles using Pannoramic Viewer (3DHISTECH, Budapest, Hungary) yielding approximately 220–1300 tiles per slide composed of 2048 × 2048 pixels each.…”

Section: Methodsmentioning

confidence: 99%

“…Digitized slides were automatically partitioned into 498 × 498 μm 2 tiles using Pannoramic Viewer (3DHISTECH, Budapest, Hungary) yielding approximately 220–1300 tiles per slide composed of 2048 × 2048 pixels each. Tiles were imported into MATLAB (MathWorks, Natick, MA) and individually converted into CIE-Lab colour space as previously described [ 14 , 15 ]. Pixels were then classified as air, tissue, oedema, or red blood cells (RBCs) using a combination of two-dimensional colour-based k -means clustering and manual clustering seed adjustment (Additional file 1 : Fig.…”

Section: Methodsmentioning

confidence: 99%

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Repeated endo-tracheal tube disconnection generates pulmonary edema in a model of volume overload: an experimental study

et al. 2022

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Background An abrupt lung deflation in rodents results in lung injury through vascular mechanisms. Ventilator disconnections during endo-tracheal suctioning in humans often cause cardio-respiratory instability. Whether repeated disconnections or lung deflations cause lung injury or oedema is not known and was tested here in a porcine large animal model. Methods Yorkshire pigs (~ 12 weeks) were studied in three series. First, we compared PEEP abruptly deflated from 26 cmH2O or from PEEP 5 cmH2O to zero. Second, pigs were randomly crossed over to receive rapid versus gradual PEEP removal from 20 cmH2O. Third, pigs with relative volume overload, were ventilated with PEEP 15 cmH2O and randomized to repeated ETT disconnections (15 s every 15 min) or no disconnection for 3 h. Hemodynamics, pulmonary variables were monitored, and lung histology and bronchoalveolar lavage studied. Results As compared to PEEP 5 cmH2O, abrupt deflation from PEEP 26 cmH2O increased PVR, lowered oxygenation, and increased lung wet-to-dry ratio. From PEEP 20 cmH2O, gradual versus abrupt deflation mitigated the changes in oxygenation and vascular resistance. From PEEP 15, repeated disconnections in presence of fluid loading led to reduced compliance, lower oxygenation, higher pulmonary artery pressure, higher lung wet-to-dry ratio, higher lung injury score and increased oedema on morphometry, compared to no disconnects. Conclusion Single abrupt deflation from high PEEP, and repeated short deflations from moderate PEEP cause pulmonary oedema, impaired oxygenation, and increased PVR, in this large animal model, thus replicating our previous finding from rodents. Rapid deflation may thus be a clinically relevant cause of impaired lung function, which may be attenuated by gradual pressure release.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Repeated endo-tracheal tube disconnection generates pulmonary edema in a model of volume overload: an experimental study

et al. 2022

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“…For each rat, between 1 and 3 5‐μm‐thick sections of both the lungs were stained with hematoxylin and eosin, then digitized on a whole‐slide scanner (Pannoramic 250 Flash II, 3DHistech, Hungary) acquiring approximately 700‐1600 micrographs per section with dimensions of 498 × 498 μm using a 20×/0.8 objective at a bright‐field resolution of 0.243 × 0.243 μm per pixel. Using MATLAB, all acquired micrographs were segmented by 4 group k‐means clustering corresponding to air, RBC, pulmonary tissue, and edema . The segmented images were binarized into RBC and non‐RBC compartments and the blood fraction was calculated as the ratio of RBC pixels to total pixels in the image.…”

Section: Methodsmentioning

confidence: 99%

“…Using MATLAB, all acquired micrographs were segmented by 4 group k-means clustering corresponding to air, RBC, pulmonary tissue, and edema. [39][40][41] The segmented images were binarized into RBC and non-RBC compartments and the blood fraction was calculated as the ratio of RBC pixels to total pixels in the image. The overall blood fraction for a rat was calculated as the mean across all segmented micrographs.…”

Section: Histologymentioning

confidence: 99%

Chemical shift of ¹²⁹Xe dissolved in red blood cells: Application to a rat model of bronchopulmonary dysplasia

Friedlander

Zanette

Lindenmaier

et al. 2019

Magnetic Resonance in Med

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Purpose To measure the chemical shift of hyperpolarized 129Xe dissolved in the red blood cells(δRBC) of a cohort of rats exposed to hyperoxia and intermittent hypoxia (IH) to mimic human bronchopulmonary dysplasia, and to investigate the effect of xenon‐blood distribution time on δRBC. Methods δRBC was measured from spectra acquired using a chemical shift saturation recovery sequence from 15 Sprague‐Dawley rats exposed to hyperoxia‐IH and 10 age‐matched control rats. Sensitization to the xenon‐blood distribution time was achieved by varying the time between saturation pulses, τ. δRBC was compared with blood fraction measured by histology of the cohort and blood oxygenation measured directly using pulse oximetry following a hypoxic challenge in an identically exposed cohort. Results The mean δRBC in the hyperoxia‐IH exposed rats was 0.55 ± 0.04 ppm lower than that of the healthy cohort (P = .0038), and this difference did not depend on τ (P = .996). The blood fraction of the exposed cohort was lower than that of the healthy cohort (P = .0397). Oximetry measurements showed that the baseline arterial oxygen saturation (SaO2) of each cohort was not different (P = .72), but after a hypoxic challenge, the SaO2 of the exposed cohort was lower than that of the healthy cohort (P = .003). Conclusion δRBC is reduced in rats exposed to hyperoxia‐IH compared with control rats. The change in δRBC is consistent with enhanced blood oxygen desaturation of the exposed cohort measured by pulse oximetry during a hypoxic challenge. This suggests that the observed change in δRBC reflects enhanced desaturation in the hyperoxia‐IH exposed cohort compared with the healthy cohort.

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Automated Whole Slide Morphometry (AWSM-Q): A Validation Study for Lung Histology Quantification in Rodent Models of Bronchopulmonary Dysplasia

Cited by 2 publications

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Repeated endo-tracheal tube disconnection generates pulmonary edema in a model of volume overload: an experimental study

Repeated endo-tracheal tube disconnection generates pulmonary edema in a model of volume overload: an experimental study

Chemical shift of ¹²⁹Xe dissolved in red blood cells: Application to a rat model of bronchopulmonary dysplasia

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Automated Whole Slide Morphometry (AWSM-Q): A Validation Study for Lung Histology Quantification in Rodent Models of Bronchopulmonary Dysplasia

Cited by 2 publications

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Repeated endo-tracheal tube disconnection generates pulmonary edema in a model of volume overload: an experimental study

Repeated endo-tracheal tube disconnection generates pulmonary edema in a model of volume overload: an experimental study

Chemical shift of 129Xe dissolved in red blood cells: Application to a rat model of bronchopulmonary dysplasia

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Chemical shift of ¹²⁹Xe dissolved in red blood cells: Application to a rat model of bronchopulmonary dysplasia