Abstract
Volumetric data at micrometer level resolution can be acquired within a few minutes using synchrotron-radiation-based tomographic microscopy. The field of view along the rotation axis of the sample can easily be increased by stacking several tomograms, allowing the investigation of long and thin objects at high resolution. On the contrary, an extension of the field of view in the perpendicular direction is non-trivial. This paper presents an acquisition protocol which increases the field of view of the tomographic dataset perpendicular to its rotation axis. The acquisition protocol can be tuned as a function of the reconstruction quality and scanning time. Since the scanning time is proportional to the radiation dose imparted to the sample, this method can be used to increase the field of view of tomographic microscopy instruments while optimizing the radiation dose for radiation-sensitive samples and keeping the quality of the tomographic dataset on the required level. This approach, dubbed wide-field synchrotron radiation tomographic microscopy, can increase the lateral field of view up to five times. The method has been successfully applied for the three-dimensional imaging of entire rat lung acini with a diameter of 4.1 mm at a voxel size of 1.48 microm.
Highlights
The functional respiratory lung unit, the so-called acinus, is defined as the complex of alveolated airways distal of a last purely conducting airway, the terminal bronchiole (Rodriguez et al, 1987)
We present a method to laterally increase the field of view of tomographic imaging systems operated in parallel-beam geometry and would like to call this method wide-field synchrotron-radiation-based X-ray tomographic microscopy (WF-SRXTM)
A method to increase the lateral field of view of tomographic imaging has been established, which enables the high-resolution tomographic imaging of large samples that are wider than the field of view of the optical set-up in multiple semi-automatically combined steps
Summary
The functional respiratory lung unit, the so-called acinus, is defined as the complex of alveolated airways distal of a last purely conducting airway, the terminal bronchiole (Rodriguez et al, 1987). The total of all acini forms the lung parenchyma, the area where the pulmonary gas-exchange takes place. While the structural development of the gas-exchange region including the alveolar septa is quite well characterized (Schittny & Burri, 2008; Schittny et al, 2008; Mund et al, 2008), the development of the three-dimensional structure of its functional unit, of the acini, has not been studied much owing to the lack of suitable methods. In order to visualize the thin sheets of tissue (alveolar septa) forming the gas-exchanging alveoli, a resolution of the order of 1 mm is required.
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