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    • 2. 发明申请
    • One-Step Cell and Tissue Preservative for Morphologic and Molecular Analysis
    • 一步细胞和组织防腐剂用于形态和分子分析
    • US20130137094A1
    • 2013-05-30
    • US13575001
    • 2011-01-25
    • Virginia A. EspinaLance A. LiottaClaudius Mueller
    • Virginia A. EspinaLance A. LiottaClaudius Mueller
    • G01N1/30
    • G01N1/30A01N1/00
    • The invention relates to a one-step chemical composition that preserves animal tissue, cells, and biomolecules, such as human tissue, human cells, and biomolecules therein. It improves the fidelity and morphologic structure of cells, organelles, and nuclear chromatin, and maintains and enhances the cellular antigenicity for immunohistochemistry and flow cytometry, while preserving proteins, post-translational modifications of proteins, and nucleic acids. In one embodiment, the composition comprises a) a non-aldehyde precipitating fixative at a concentration below 25% (volume/volume), b) a reversible/cleavable protein cross-linker that targets lipid-associated molecules, and c) a c reversible/cleavable protein cross-linker that targets water soluble molecules. In another embodiment, the composition further includes a kinase inhibitor, a phosphatase inhibitor, and a permeation enhancer. In still another embodiment, the compositions further include lactic acid at a concentration sufficient to maintain cellular nuclear volume at a level equivalent to aldehyde fixation of the same type of cell. In a further embodiment, the composition comprises: a) a precipitating fixative, b) a reversible/cleavable cross-linker, c) a permeation enhancer, d) a kinase inhibitor, e) a phosphatase inhibitor, and f) a carboxylic acid. In a still further embodiment, the invention comprises method for preserving a biological sample by contacting the sample with the composition of the invention under conditions effective for the preservation of the sample.
    • 本发明涉及保留动物组织,细胞和生物分子的一步化学组合物,例如人组织,人细胞和生物分子。 它提高细胞,细胞器和核染色质的保真度和形态结构,并保持和增强免疫组织化学和流式细胞术的细胞抗原性,同时保留蛋白质,蛋白质的翻译后修饰和核酸。 在一个实施方案中,组合物包含a)浓度低于25%(体积/体积)的非醛沉淀固定剂,b)靶向脂质相关分子的可逆/可切割蛋白质交联剂,以及c)可逆可逆/ 可裂解的蛋白质交联剂,其靶向水溶性分子。 在另一个实施方案中,组合物还包含激酶抑制剂,磷酸酶抑制剂和渗透增强剂。 在另一个实施方案中,组合物还包括浓度足以将细胞核体积保持在与相同类型细胞的醛固定相当的水平的乳酸。 在另一个实施方案中,组合物包括:a)沉淀固定剂,b)可逆/可切割交联剂,c)渗透促进剂,d)激酶抑制剂,e)磷酸酶抑制剂,和f)羧酸。 在又一个实施方案中,本发明包括通过在有效保存样品的条件下使样品与本发明的组合物接触来保存生物样品的方法。