A Simple, Inexpensive, and Effective Light- Carrying Laryngoscopic Blade for Orotracheal Intubation of Rats

The research paradigm of using large laboratory animals, in which oroendotracheal intubations are relatively easy, is shifting toward the use of small animals, such as rodents, in which oropharyngeal access is limited, the arytenoid cartilage cycles are faster, and the glottis is much smaller. The considerable growth recently seen in preclinical imaging studies is accompanied by an increased number of rats and mice requiring in vivo intubation for airway management. Tracheal access is important for ventilation, administration of inhaled anesthetics, instillation of drugs or imaging agents, and maintenance of airway patency to reduce mortality during and after operations. I fashioned a light-carrying laryngoscopic blade (laryngoscope) from readily available acrylic-polymethyl methacrylate tubing and used it to perform rapid, effective tracheal intubation in rats. The laryngoscope design and intubation techniques are presented

Automation Process for Morphometric Analysis of Volumetric CT Data from Pulmonary Vasculature in Rats

With advances in medical imaging scanners, it has become commonplace to generate large multidimensional datasets. These datasets require tools for a rapid, thorough analysis. To address this need, we have developed an automated algorithm for morphometric analysis incorporating A Visualization Workshop computational and image processing libraries for three-dimensional segmentation, vascular tree generation and structural hierarchical ordering with a two-stage numeric optimization procedure for estimating vessel diameters. We combine this new technique with our mathematical models of pulmonary vascular morphology to quantify structural and functional attributes of lung arterial trees. Our physiological studies require repeated measurements of vascular structure to determine differences in vessel biomechanical properties between animal models of pulmonary disease. Automation provides many advantages including significantly improved speed and minimized operator interaction and biasing. The results are validated by comparison with previously published rat pulmonary arterial micro-CT data analysis techniques, in which vessels were manually mapped and measured using intense operator intervention

The Effect of ACE Inhibition on the Pulmonary Vasculature in Combined Models of Chronic Hypoxia and Pulmonary Arterial Banding in Sprague Dawley Rats

Microfocal CT was used to image the pulmonary arterial (PA) tree in rodent models of pulmonary hypertension (PH). CT images were used to measure the arterial tree diameter along the main arterial trunk at several hydrostatic intravascular pressures and calculate distensibility. High-resolution planar angiographic imaging was also used to examine distal PA microstructure. Data on pulmonary artery tree morphology improves our understanding of vascular remodeling and response to treatments. Angiotensin II (ATII) has been identified as a mediator of vasoconstriction and proliferative mitotic function. ATII has been shown to promote vascular smooth muscle cell hypertrophy and hyperplasia as well as stimulate synthesis of extracellular matrix proteins. Available ATII is targeted through angiotensin converting enzyme inhibitors (ACEIs), a method that has been used in animal models of PH to attenuate vascular remodeling and decrease pulmonary vascular resistance. In this study, we used rat models of chronic hypoxia to induce PH combined with partial left pulmonary artery occlusion (arterial banding, PLPAO) to evaluate effects of the ACEI, captopril, on pulmonary vascular hemodynamic and morphology. Male Sprague Dawley rats were placed in hypoxia (FiO2 0.1), with one group having underwent PLPAO three days prior to the chronic hypoxia. After the twenty-first day of hypoxia exposure, treatment was started with captopril (20 mg/kg/day) for an additional twenty-one days. At the endpoint, lungs were excised and isolated to examine: pulmonary vascular resistance, ACE activity, pulmonary vessel morphology and biomechanics. Hematocrit and RV/LV+septum ratio was also measured. CT planar images showed less vessel dropout in rats treated with captopril versus the non-treatment lungs. Distensibility data shows no change in rats treated with captopril in both chronic hypoxia (CH) and CH with PLPAO (CH+PLPAO) models. Hemodynamic measurements also show no change in the pulmonary vascular resistance with captopril treatment in both CH and CH+PLPAO

Post-Acquisition Small-Animal Respiratory Gated Imaging Using Micro Cone-Beam CT

On many occasions, it is desirable to image lungs in vivo to perform a pulmonary physiology study. Since the lungs are moving, gating with respect to the ventilatory phase has to be performed in order to minimize motion artifacts. Gating can be done in real time, similar to cardiac imaging in clinical applications, however, there are technical problems that have lead us to investigate different approaches. The problems include breath-to-breath inconsistencies in tidal volume, which makes the precise detection of ventilatory phase difficult, and the relatively high ventilation rates seen in small animals (rats and mice have ventilation rates in the range of a hundred cycles per minute), which challenges the capture rate of many imaging systems (this is particularly true of our system which utilizes cone-beam geometry and a 2 dimensional detector). Instead of pre-capture ventilation gating we implemented a method of post-acquisition gating. We acquire a sequence of projections images at 30 frames per second for each of 360 viewing angles. During each capture sequence the rat undergoes multiple ventilation cycles. Using the sequence of projection images, an automated region of interest algorithm, based on integrated grayscale intensity, tracts the ventilatory phase of the lungs. In the processing of an image sequence, multiple projection images are identified at a particular phase and averaged to improve the signal-to-ratio. The resulting averaged projection images are input to a Feldkamp cone-beam algorithm reconstruction algorithm in order to obtain isotropic image volumes. Minimal motion artifact data sets improve qualitative and quantitative analysis techniques useful in physiologic studies of pulmonary structure and function

Quasi-Exact Helical Cone Beam Reconstruction for Micro CT

A cone beam micro-CT system is set up to collect truncated helical cone beam data. This system includes a micro-focal X-ray source, a precision computer-controlled X-Y-Z-theta stage, and an image-intensifier coupled to a large format CCD detector. The helical scanning mode is implemented by rotating and translating the stage while keeping X-ray source and detector stationary. A chunk of bone and a mouse leg are scanned and quasi-exact reconstruction is performed using the approach proposed in J. Hu et al. (2001). This approach introduced the original idea of accessory paths with upper and lower virtual detectors having infinite axial extent. It has a filtered backprojection structure which is desirable in practice and possesses the advantages of being simple to implement and computationally efficient compared to other quasi-exact helical cone beam algorithms for the long object problem

Measuring the effect of airway pressure on pulmonary arterial diameter in the intact rat lung

To study the relationship between transpulomnary pressure (Ptp), intravascular pressure (Pv), and the pulmonary arterial tree structure, morphometric measurements of pulmonary arterial trees were made in intact lungs from Sprague-Dawley rats. Using cone beam micro-CT and techniques we developed for imaging small animal lungs, volumetric CT data were acquired for Ptp from 0 - 12 mmHg and Pv from 5 - 30 mmHg. The diameter, D (measured range approximately 0.08-2.0 mm), vs. pressure, P, relation can be described by D(P) = D(0)(1+ α P), where α is a distensibility coefficient. Unlike studies performed in larger animals, where changes in either Ptp or Pv had nearly identical effect on vessel distensibility, we found that there is only a small dependence of arterial diameter on Ptp in the rat. For example, using the above relation where P=Ptp and Pv is held constant at 12mmHg, alpha = 0.55±0.42(SE) %/mmHg, compared with when P=Pv and Ptp is held at 12mmHg, alpha = 2.59±0.17(SE) %/mmHg

Quantification of Pulmonary Arterial Wall Distensibility Using Parameters Extracted from Volumetric Micro-CT Images

Stiffening, or loss of distensibility, of arterial vessel walls is among the manifestations of a number of vascular diseases including pulmonary arterial hypertension. We are attempting to quantify the mechanical properties of vessel walls of the pulmonary arterial tree using parameters derived from high-resolution volumetric x-ray CT images of rat lungs. The pulmonary arterial trees of the excised lungs are filled with a contrast agent. The lungs are imaged with arterial pressures spanning the physiological range. Vessel segment diameters are measured from the inlet to the periphery, and distensibilities calculated from diameters as a function of pressure. The method shows promise as an adjunct to other morphometric techniques such as histology and corrosion casting. It possesses the advantages of being nondestructive, characterizing the vascular structures while the lungs are imaged rapidly and in a near-physiological state, and providing the ability to associate mechanical properties with vessel location in the intact tree hierarchy