Core--Analytical laboratory

这一核心设施为项目1、2和3提供分析化学和组织病理学支持。提供的神经化学分析包括在血清、脑脊液和微透析液样本中测量的血管活性化合物和代谢物。用高效液相色谱法测定一氧化氮相关氨基酸精氨酸和瓜氨酸，兴奋性氨基酸谷氨酸和天冬氨酸，体积调节性氨基酸牛磺酸和抑制性氨基酸甘氨酸。在生物能量窘迫时升高的血管活性嘌呤-腺苷也被用高效液相色谱法测定。微透析液中的钾随着膜的丧失而升高，膜电位的维持性用离子色谱法测定。用化学发光法测定了一氧化氮、硝酸盐和亚硝酸盐的主要分解产物。血管活性多肽，如内皮素，用酶联免疫测定法测定。组织学支持包括照片和数字文件、大体检查、组织处理、组织切片和染色。用图像分析软件进行挫伤体积和神经细胞计数。使用毛细血管灌流的腔内标志物和毛细血管内皮细胞的免疫组织化学标志物，将测量灌流的毛细血管的比例，并根据需要进行免疫组织化学染色；目前的免疫组织化学染色包括β-淀粉样蛋白、葡萄糖和单羧酸转运体、内皮素、一氧化氮合酶(NOS)、凝血因子8和3-硝基酪氨酸。

This core facility provides analytical chemistry and histopathology support for Projects 1,2, and 3. The neurochemical analysis provided include vasoactive compounds and metabolites measured in serum, cerebrospinal fluid, and microdialysate samples. Amino acids including the nitric oxide related amino acids arginine and citrulline, the excitatory amino acids glutamate and aspartate, the volume regulatory amino acid taurine and the inhibitory amino acid glycine are measured by high performance liquid chromatography. The vasoactive purine adenosine which is elevated during bioenergetic distress is also measured by HPLC. Potassium in microdialysate is elevated with loss of membrane potential maintenance is measured by ion chromatography. The major breakdown products of nitric oxide, nitrate and nitrite, are measured using chemiluminescence. Vasoactive peptides such as endothelin are measured using enzyme- linked immunoassay. Histology support consists of photographic and digital documentation, gross examination, tissue processing, tissue sectioning, and staining. Contusion volume and neuronal cell counting is performed using image analysis software. Using intralumenal markers of capillary perfusion and immunohistochemical markers of capillary endothelial cells, the fraction of perfused capillaries will be measured morphologenically Immunohistochemical staining an also be performed as need; the current repertoire of immunohistochemical staining includes beta-amyloid, glucose and monocarboxylate transporters, endothelin, nitric oxide synthetase (NOS), coagulation factor 8, and 3-nitrotyrosine.