Biomedical subjects

Chantal Rémy

Publications and source records attributed to Chantal Rémy.

9 recordsLinked to original sources

PubMed2006

A direct method for measuring mouse capillary cortical blood volume using multiphoton laser scanning microscopy.

Knowledge of the blood volume per unit volume of brain tissue is important for understanding brain function in health and disease. We describe a direct method using two-photon laser scanning microscopy to obtain in vivo the local capillary blood volume in the cortex of anesthetized mouse. We infused fluorescent dyes in the circulating blood and imaged the blood vessels, including the capillaries, to a depth of 600 microm below the dura at the brain surface. Capillary cortical blood volume (CCBV) was calculated without any form recognition and segmentation, by normalizing the total fluorescence measured at each depth and integrating the collected intensities all over the stack. Theoretical justifications are presented and numerical simulations were performed to validate this method which was weakly sensitive to background noise. Then, CCBV had been estimated on seven healthy mice between 2%+/-0.3% and 2.4%+/-0.4%. We showed that this measure of CCBV is reproductible and that this method is highly sensitive to the explored zones in the cortex (vessel density and size). This method, which dispenses with form recognition, is rapid and would allow to study in vivo temporal and highly resolute spatial variations of CCBV under different conditions or stimulations.

Algorithms↗

PubMed2006

A low temperature embedding and section registration strategy for 3D image reconstruction of the rat brain from autoradiographic sections.

In studies on animal models of human brain pathologies, three-dimensional reconstruction from histological sections is particularly useful when assessing the morphologic, functional and biochemical changes induced by pathology. It allows assessing lesion heterogeneity in planes different from the cutting plane and allows correlating the histology with images obtained in vivo, such as by means of magnetic resonance imaging. To create a 3D volume from autoradiographic sections with minimal distortion, both cryosectioning as well as section registration need to be optimal. This paper describes a strategy whereby four external fiducial markers are positioned outside the rat brain with the use of a low temperature brain embedding procedure. The fiducial markers proposed here can be rapidly added to any frozen tissue block with no impact on the subsequent histological operations. Since embedding is performed at a low temperature, no tissue degradation occurs due to sample heating. The markers enable robust and almost error free registration, even in the presence of missing sections and poor image quality. Furthermore, the markers may be used to partially correct for global distortions.

Acrylamides↗

PubMed2006

In vivo two-photon microscopy study of short-term effects of microbeam irradiation on normal mouse brain microvasculature.

PURPOSE: The purpose of this study was to assess the early effects of microbeam irradiation on the vascular permeability and volume in the parietal cortex of normal nude mice using two-photon microscopy and immunohistochemistry. METHODS AND MATERIALS: The upper part of the left hemisphere of 55 mice was irradiated anteroposteriorly using 18 vertically oriented beams (width 25 microm, interdistance 211 microm; peak entrance doses: 312 or 1000 Gy). At different times after microbeam exposure, the microvasculature in the cortex was analyzed using intravital two-photon microscopy after intravascular injection of fluorescein isothiocyanate (FITC)-dextrans and sulforhodamine B (SRB). Changes of the vascular volume were observed at the FITC wavelength over a maximum depth of 650 mum from the dura. The vascular permeability was detected as extravasations of SRB. RESULTS: For all times (12 h to 1 month) after microbeam irradiation and for both doses, the FITC-dextran remained in the vessels. No significant change in vascular volume was observed between 12 h and 3 months after irradiation. Diffusion of SRB was observed in microbeam irradiated regions from 12 h until 12 days only after a 1000 Gy exposure. CONCLUSION: No radiation damage to the microvasculature was detected in normal brain tissue after a 312 Gy microbeam irradiation. This dose would be more appropriate than 1000 Gy for the treatment of brain tumors using crossfired microbeams.

Animals↗

PubMed2006

Development of a decision support system for diagnosis and grading of brain tumours using in vivo magnetic resonance single voxel spectra.

A computer-based decision support system to assist radiologists in diagnosing and grading brain tumours has been developed by the multi-centre INTERPRET project. Spectra from a database of 1H single-voxel spectra of different types of brain tumours, acquired in vivo from 334 patients at four different centres, are clustered according to their pathology, using automated pattern recognition techniques and the results are presented as a two-dimensional scatterplot using an intuitive graphical user interface (GUI). Formal quality control procedures were performed to standardize the performance of the instruments and check each spectrum, and teams of expert neuroradiologists, neurosurgeons, neurologists and neuropathologists clinically validated each case. The prototype decision support system (DSS) successfully classified 89% of the cases in an independent test set of 91 cases of the most frequent tumour types (meningiomas, low-grade gliomas and high-grade malignant tumours--glioblastomas and metastases). It also helps to resolve diagnostic difficulty in borderline cases. When the prototype was tested by radiologists and other clinicians it was favourably received. Results of the preliminary clinical analysis of the added value of using the DSS for brain tumour diagnosis with MRS showed a small but significant improvement over MRI used alone. In the comparison of individual pathologies, PNETs were significantly better diagnosed with the DSS than with MRI alone.

Algorithms↗

PubMed2005

Focal brain ischemia in rat: acute changes in brain tissue T1 reflect acute increase in brain tissue water content.

Several recent studies have reported changes of brain tissue T(1) in ischemic models during the first minutes after occlusion of the middle cerebral artery (MCA). In order to assess whether these tissue T(1) changes are related to an increase in tissue water content, we performed T(1) (7 T) and tissue water content measurements in a rat model (n = 10, Sprague-Dawley) of focal cerebral ischemia (intraluminal occlusion model). The tissue water content was determined using a gravimetric technique. The animals were divided into two groups: an ischemic group, with an effective MCA occlusion (n = 6) and a control group, with animals having undergone sham surgery but no MCA occlusion (n = 4). In the ipsilateral cortex, the tissue water content was 81.1 +/- 0.7% at 2 h 15 min following ischemic insult (contralateral value: 79.3 +/- 0.5%). Concomitantly, the tissue T(1) in the ipsilateral cortex was 2062 +/- 60 ms at ischemia onset + 1 h (contralateral 1811 +/- 28 ms) and 2100 +/- 38 ms at ischemia onset + 2 h (contralateral 1807 +/- 18 ms). The tissue T(1) and tissue water content values measured in the contralateral area do not differ from the values obtained in the control group. A significant T(1) increase is observed at ischemia onset + 1 h (+ 14%) and ischemia onset (+ 2 h) + 16%, together with a significant increase in tissue water content (+ 2.3%). This suggests that there is an increase in tissue water content concomitant with cell swelling during the first hours of ischemia.

Animals↗

PubMed2004

2D J-resolved spiral spectroscopic imaging at 7 T: application to mobile lipid mapping in a rat glioma.

Lactate (Lac) and mobile lipids (Lip), which are present in rat gliomas, are difficult to map because their 1H resonances overlap in the 1.3 ppm region. 2D J-resolved spectroscopy enables proper separation of the two resonance lines. To obtain high-spatial-resolution mapping of Lac and Lip resonances within a reasonable experiment time, we coupled 2D J-resolved spectroscopy with a fast spectroscopic imaging (SI) method, based on an out-and-in spiral k-space description. The method was applied to a rat glioma at 7 T, and Lac and Lip maps were reconstructed. The duration of SI (2D spatial, 2D spectral) was 64 min for a theoretical in-plane resolution of 1 x 1 mm, and a slice thickness of 2 mm (voxel size 8.2 microl, taking into account the point-spread function (PSF)).

Animals↗

PubMed2003

Correlation between the occurrence of 1H-MRS lipid signal, necrosis and lipid droplets during C6 rat glioma development.

The aim of this study was to investigate the possible correlation between the 1H MRS mobile lipid signal, necrosis and lipid droplets in C6 rat glioma. First, the occurrence of necrosis and lipid droplets was determined during tumor development, by a histological analysis performed on 34 rats. Neither necrosis nor lipid droplets were observed before 18 days post-implantation. At later stages of development, both necrosis and lipid droplets were apparent, the lipid droplets being mainly located within the necrotic areas. Using a second group of eight rats, a temporal correlation was evidenced between mobile lipid signal detected by in vivo single-voxel one- (136 ms echo time) and two-dimensional J-resolved 1H MR spectroscopy, and the presence of necrosis and lipid droplets on the histological sections obtained from the brains of the same rats. Finally, spatial distribution of the mobile lipid signal was analyzed by chemical-shift imaging performed on a third group of eight animals, at the end of the tumor growth. The spectroscopic image corresponding to the resonance of mobile lipids had its maximum intensity in the center of the tumor where necrotic regions were observed on the histological sections. These necrotic areas contained large amounts of lipid droplets. All these results suggest that mobile lipids detected in vivo by 1H MRS (136 ms echo time) in C6 rat brain glioma arise mainly from lipid droplets located in necrosis.

Animals↗

PubMed2002

Measurement by nuclear magnetic resonance diffusion of the dimensions of the mobile lipid compartment in C6 cells.

The (1)H spectrum of certain tumor cells, in vivo tumors, and their biopsies in vitro shows a narrow and intense resonance at 1.26 ppm, which has been assigned to the fatty acyl chain of triglycerides [nuclear magnetic resonance (NMR) visible mobile lipids, MLs]. We have used diffusion-weighted NMR spectroscopy to directly address the subcellular origin of MLs in the case of C6 cells in which lactate accumulation had been inhibited by prior iodoacetamide incubation. Borage oil and artificial lipid droplets were used as model systems of free and restricted diffusion, respectively. The characteristic diameter for the ML resonance compartment measured by NMR for the C6 cells was not significantly different from the one obtained with phase contrast microscopy (1.88 +/- 0.04 micro m from NMR versus 1.37 +/- 0.33 micro m from microscopy). We herewith provide direct and noninvasive evidence that the lipid signal at 1.26 ppm in C6 cells, which remains visible in long echo time (T(E) = 136 ms) experiments, mostly originates from subcellular structures with diameters of 1-2 micro m, which correspond to the cytosolic lipid droplets that can be detected in optical microscopy preparations of the same cells.

Animals↗

PubMedDate not supplied

Immunohistochemical study of VEGF, angiopoietin 2 and their receptors in the neovascularization following microinjection of C6 glioma cells into rat brain.

BACKGROUND: Recent papers suggest that two angiogenic factors (angiopoietin 2 and vascular endothelial growth factor) cooperate in tumoral angiogenesis to support the growing tumor. The purpose of the present work was to demonstrate the existence of such cooperation in a longitudinal study of a brain tumor model during tumor growth by means of immunohistochemistry. MATERIALS AND METHODS: The study was performed on 31 rats bearing C6 glioma. At different stages of tumor growth, the histological aspects were described and sections were immunostained for VEGF, Ang-2 and their receptors VEGFR-1, VEGFR-2 and Tie-2. Immunostaining was semi-quantitatively analyzed and the localization of immunostained cells was reported. RESULTS: Ang-2 and Tie-2 were detected in the endothelial cells of vessels surrounded by tumor cells, occuring early in our study, with immunostaining taking place from day 4 to day 24. Immunostaining with VEGF (on tumoral cells) and VEGFR-2 (on endothelial cells) was present after 8 days of tumor growth. A clear increase of vessel density can be observed at the periphery of the tumors after 16 days of tumor growth. At that time, areas of necrosis were present in the tumor with concomitant VEGF and Ang-2 expression. CONCLUSION: The present study demonstrated cooperation between the early effect of Ang-2 and the secondary effect of VEGF to elaborate new vessels in a longitudinal study of experimental brain tumors. This study is favorable to the new model of tumor angiogenesis, with successive vessel cooption, regression and growth mediated by angiopoietins and VEGF.

Angiopoietin-2↗