Toshihiro Sera

Three-dimensional visualization and morphometry of small airways from microfocal X-ray computed tomography.

Physiological morphometry is a critical factor in the flow dynamics in small airways. In this study, we visualized and analyzed the three-dimensional structure of the small airways without dehydration and fixation. We developed a two-step method to visualize small airways in detail by staining the lung tissue with a radiopaque solution and then visualizing the tissue with a cone-beam microfocal X-ray computed tomographic (CT) system. To verify the applicability of this staining and CT imaging (SCT) method, we used the method to visualize small airways in excised rat lungs. By using the SCT method to obtain continuous CT images, three-dimensional branching and merging bronchi ranging from 500 to 150 microm (the airway generation=8-16) were successfully reconstructed. The morphometry of the small airways (diameter, length, branching angle and gravity angle between the gravity direction and airway vector) was analyzed using the three-dimensional thinning algorithm. The diameter and length exponentially decreased with the airway generation. The asymmetry of the bifurcation decreased with generation and one branching angle decided the other pair branching angle. The SCT method is the first reported method that yields faithful high-resolution images of soft tissue geometry without fixation and the three-dimensional morphometry of small airways is useful for studying the biomechanical dynamics in small airways.

Development of high-resolution 4D in vivo-CT for visualization of cardiac and respiratory deformations of small animals.

The interest in small animal models of human diseases has generated a need to design a computed tomography (CT) system that operates at a microscopic level. It is particularly important to be able to visualize the dramatic rhythmical motion of organs such as the heart and lungs. In order to evaluate the motion of the heart and lungs of small animals (rats and mice), we developed in the present study a high-resolution 4D in vivo-CT system for small animals that uses synchrotron radiation. To reduce motion artifacts and the radiation dose, the projections were synchronized with airway pressure, the ECG, the x-ray shutter and the CCD shutter. For cardiovascular imaging, a blood pool contrast agent was injected and the data sets were acquired at several ECG points during the end-expiratory phase. For imaging of the lungs, the data sets were acquired at several airway pressures during diastole. The dynamic motion of the cardiovascular system (the ventricles and coronary arteries) and small airways (diameter > 250 microm of rats and 125 microm of mice) was visualized. This high-resolution imaging tool may be very useful for the development of novel drugs in murine models, in addition to its use in the study of cardiovascular and respiratory physiology.

Fast tomography using quasi-monochromatic undulator radiation.

A beamline with a helical undulator has been used without a monochromator for fast high-resolution tomographic imaging with an X-ray energy of 12.4-16.5 keV and an energy bandwidth of 2-3%. The X-ray beam was expanded with two mirrors to 12 mm x 4 mm. The X-ray field was made uniform by a diffuser. The detector pixel size was 9.9 microm x 9.9 microm. At the highest speed, a 180 degrees scan was completed in 6 s with 454 projections. Beam-hardening effects were not significant. This technique may be useful in studying time-dependent structural changes of soft materials such as polymers and biological samples.

Small airway changes in healthy and ovalbumin-treated mice during quasi-static lung inflation.

Previously, we developed a synchrotron radiation CT system to evaluate the morphometric changes (length and diameter, D) and small airway compliance (sC(aw)) of euthanized mice under quasi-static inflation [Sera, T., Uesugi, K., Yagi, N., 2005. Localized morphometric deformations of small airways and alveoli in intact mouse lungs under quasi-static inflation. Respir. Physiol. Neurobiol. 147, 51-63). Using this system, this study compared normal and asthmatic small airways. Ovalbumin-treated mice were used as an asthma model. Compared with the values at functional residual capacity, D of normal and asthmatic small airways (D<200microm) increased by 48% and 36% at the end of tidal inspiration. For larger airways (D>500microm), the increases were 23% and 20%, respectively. The ratio of the sC(aw) of asthmatic small airways to that of healthy small airways was 0.57, and the ratio was 0.70 for larger airways. The morphometric changes and sC(aw) in asthma model mice were significantly lower than those of healthy mice. The differences in sC(aw) between healthy and asthma model mice were greater for smaller airways.

Localized morphometric deformations of small airways and alveoli in intact mouse lungs under quasi-static inflation

Localized morphometric deformations of small airways and alveoli during respiration have several biomechanical and physiological implications. We developed fast synchrotron radiation CT system to visualize the small airways and alveoli of an intact mouse lung without fixation and dehydration, and analyzed their localized morphometric deformations between functional residual capacity (FRC) and total lung capacity (TLC). The maximum resolution of 32.6lp/mm at the 5% modulation transfer function level can be achieved with 11.8-microm voxels and 7-min scanning. Compared with the values at FRC, the diameter and length for smaller airways (diameter at FRC <200 microm) increased by 68.8% and 29.5% (averaged value), and those for larger airways (diameter at FRC >400 microm) increased by 45.2 and 22.9% (averaged value), at TLC. Moreover we defined the volume behavior as the percentage of airway volume at FRC for TLC. The volume behavior for the small airways was not similar to that of the lung volume. These results indicated that all airways did not behave homogenously.

High-resolution visualization of tumours in rabbit lung using refraction contrast X-ray imaging.

Contrast enhancement by refraction was used to visualize tumours in the rabbit lung. VX2 tumour cells were intravenously injected into a rabbit. After 14 days the rabbit was euthanized and the lungs were imaged. Refraction-enhanced X-ray images were obtained with a sample-to-detector distance of 2.65-6m. The beamline BL20B2 at the SPring-8 synchrotron radiation facility was used for the experiment, with a monochromatic X-ray beam with an energy of 33.2 keV. In the case of projection images, it was found that refraction did not help visualization of small tumours: the nodules did not show up with sharply defined edges. In tomography, tumours with a size of 1-10mm were clearly visualized, together with blood vessels with a diameter down to 0.4mm. These results show that refraction-enhanced imaging may be useful in human lung tomography to find small tumours.

In vivo-CT system with respiratory and cardiac gating using synchrotron radiation

The interest in using small animal models of human disease has produced a need to design a CT system at a microscopic level comparable to that achievable in a clinical CT in human. In this study, we developed a high-resolution in vivo-CT system with respiratory and cardiac gating using synchrotron radiation. The system was constructed in BL20B2 at SPring-8. SPring-8 is the third generation synchrotron radiation source in Hyogo, Japan, and prefers much higher flux X-ray than a laboratory X-ray source. Another advantage of synchrotron monochromatic CT is the minimalization of beam hardening effects, which pose serious problems when using white X-rays. Since the X-ray beam from the synchrotron source is parallel, each horizontal line corresponds to a slice position along the rotation axis. Multiple slices are easily obtained in one rotation (3D-CT). For in vivo scanning, the X-ray mechanical shutter and CCD electrical shutter were synchronized with an airway pressure (respiratory) and electrocardiographic (ECG) signal. Synchronization reduced the motion artifacts caused by respiration and heart beats, markedly improving visualization of the edges of the heart, ribs and diaphragm. In particular, small airways (diameter > 300 &mgr;m) and cerebral blood vessels were visualized clearly. This system is very useful for evaluating lung physiology and cardiovascular mechanics in vivo.

Development of 4-D High-Resolution CT for Measurement of Airway Deformation

Airway consists of a number of various compliant tubes from trachea to alveolus, and the airway geometry deforms dramatically during respiration. This morphometric deformation is very important factors in pulmonary dynamics (flow structure, particle deposition, surfactant transport and so on). Previously, we reported the small airway deformations of euthanized animals using micro-CT and synchrotron radiation-CT.Copyright