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    • 9. 发明公开
    • Cooperating capillary and cap for use in a high-pressure freezer
    • ZusammenwirkendesKapillarröhrchenund Verschluss zur Verwendung in einemHochdrucktiefkühlgerät
    • EP2706342A1
    • 2014-03-12
    • EP12183821.3
    • 2012-09-11
    • FEI COMPANY
    • Schampers, RuudPersoon, HansEngelen, Andre
    • G01N1/42
    • G01N1/30G01N1/42
    • Assembly of cooperating capillary (1) and cap (2) for containing an aqueous solution (3) in an inner volume of the capillary, the assembly equipped to be used in a high pressure freezer in which the aqueous solution is frozen at a high pressure to form an amorphous frozen sample at a cryogenic temperature, the inner volume of the capillary having a diameter of less than 500 µm, preferably less than 350 µm, and the cap equipped to form a closure to one end of the capillary, characterized in that the part of the cap that is in contact with the inner volume of the capillary when the cap forms a closure, shows an indent, as a result of which the cap, after freezing the aqueous solution, can be removed from the capillary and a free standing pillar of frozen aqueous material extends from the capillary.
      As a result of this the metal of the capillary need not be cut and related artifacts are eliminated. Further inspection is preferably done using a focused ion beam forming block-face or lamella and an electron beam for the inspection of the sample.
    • 用于在毛细管的内部容积中含有水溶液(3)的配合毛细管(1)和盖(2)的组装,该组件装备用于高压冷冻机中,其中水溶液在高压下冷冻 在低温下形成无定形冷冻样品,毛细管的内部体积具有小于500μm,优选小于350μm的直径,并且帽被设置成在毛细管的一端形成封闭物,其特征在于, 当盖形成闭合件时,与毛细管的内部容积接触的帽的部分显示凹口,结果是在冷冻水溶液之后,帽可以从毛细管中除去,并且自由 冷冻水性材料的立柱从毛细管延伸。 作为其结果,毛细管的金属不需要被切割并消除相关的伪像。 优选使用聚焦离子束形成阻挡面或薄片和用于检查样品的电子束进行进一步的检查。
    • 10. 发明公开
    • Forming an electron microscope sample from high-pressure frozen material
    • 用高压冷冻材料形成电子显微镜样品
    • EP2685233A1
    • 2014-01-15
    • EP12176254.6
    • 2012-07-13
    • FEI COMPANY
    • Schampers, RuudHayles, MikeDe Winter, MatthijsSchneijdenberg, Chris
    • G01N1/42
    • B23K15/08G01N1/06G01N1/42G01N2001/2873
    • Method of forming a sample from a capillary with high-pressure frozen sample material, the method comprising: a step (102) of providing a high-pressure capillary with vitrified sample material at a temperature T 1 below the glass transition temperature Tg, and a step (104) of cutting the capillary, characterized by a step (106) of warming the capillary to a temperature T 2 between temperature T 1 and temperature Tg, a subsequent step (108) of cooling the capillary to a temperature T 3 below temperature T 2 , as a result of which material is extruded from the capillary, and a step (112) of freeing a sample from the extruded sample material at a temperature below temperature T g . Vitrified biological material could be extruded from a capillary at liquid at a temperature of T = 88 K (liquid nitrogen by raising the temperature to, for example 130 K and then cooling the capillary to 88 K. Not only did this result to extrusion material, but even more surprising is that a repeated temperature cycle often results in further extrusion of sample material.
    • 用高压冷冻样品材料从毛细管形成样品的方法,所述方法包括:在低于玻璃化转变温度Tg的温度T1下向玻璃化样品材料提供高压毛细管的步骤(102);以及步骤 (104),其特征在于将毛细管加热至温度T1与温度Tg之间的温度T2的步骤(106),将毛细管冷却至低于温度T2的温度T3的后续步骤(108),作为 从毛细管挤出材料的结果,以及在低于温度Tg的温度下从挤出的样品材料中释放样品的步骤(112)。 玻璃化的生物材料可以在温度为T = 88K的液体(液氮通过将温度升高至例如130K,然后将毛细管冷却至88K)从毛细管挤出。不仅产生挤出材料, 但更令人惊讶的是,重复的温度循环常常导致样品材料的进一步挤出。