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蛋白晶体取放工具MicroMounts™
MicroMounts™是晶体学(蛋白质晶体、病毒晶体、小分子、无机晶体等)实验室常用显微工具之一,也是传统的和高通量的X射线晶体实验中的理想操作工具,例如,可以用来从溶液中取出目标晶体移到X射线通路上。它的特殊材质和设计既可视且X光可透、具有一定的机械硬度、一定的柔韧度、精准的尺寸。
这些显微工具的头部由柔韧的微纤维聚酰亚胺薄膜制造,这种柔中带刚的工具*易弯曲。这些工具可以测量样品的范围在毫米到几个微米之间,与金属工具相比,聚酰亚胺膜类工具对于很脆蛋白质晶体样品可以达到*低的损伤度,且可视和X光可透。
每个聚酰亚胺膜头部都镶嵌在一个无磁不锈钢针座(pins)上,不锈钢针座的直径大约0.64mm。针座再装配到一直径为0.7mm的手柄上,每个工具包都有手柄。该工具的应用领域:蛋白结晶学、单细胞以及较小样品。
特性:
· 从同步加速器中或者其它小型射线源中方便快速的找到拿取50μm-100μm以下尺寸的晶体
· 从溶液中找到和拿取晶体
· **拿取的蛋白质尺寸
· 每次**放置在同一位置、X射线束下简单排列
· *小化目标晶体周围的液体,以得到*佳冷却速率
· *小化背景信号使得X射线信号*大化
一、聚酰亚胺膜的模式:
1.晶体孔Crystal aperture,图示A ,大小从10-600μm,孔边缘的聚酰亚胺膜(图中B)很窄,以小化各个方向上的散射背景。
2.毛细作用孔Wicking hole,图中C,通过*细的导泄道drainage channel(图中D)与晶体孔连接。孔的大小与15 or XF paper wicks相当。钢笔样式的设计可以很好的去除多余的液体而不用the wick去接触晶体。
3.十字孔Alignment cross,图中E,位于从晶体孔中心对应的一个固定点上,便于晶体放置时的自动排列。
4.晶体孔大小代码Crystal aperture size code,图中F,围绕十字孔分布,可以用来自动识别孔的模式和晶体的大小。
5.晶体孔大小Aperture size,图中G,微米大小,直接读出。
6.方位标记孔Orientation mark ,图中H,可以用来自动识别样品在正面还是背面。
聚酰亚胺膜参数如下表所示:
Material | Polyimide (used in Kapton tape) |
Crystal aperture sizes | 10, 20, 30, 50, 75, 100, 150, 200, 300, 400, 500 and 600 micrometers |
Thickness | 7.5, 10 and 12.5 micrometers |
Distance from center of alignment cross to center of crystal aperture | 870 micrometers |
Distance from pin base to center of crystal aperture | 12.1 mm (for 11 mm pins) |
Width of wicking hole | 250 micrometers. Compatible with Size 15 paper wicks. |
Length tolerance | 0.5 mm for 11, 19 and 25 mm pins 0.3 mm for SPINE pins. |
二、无磁不锈钢针座Solid 316 non-magnetic stainless steel pins.
提供的的针座有四种规格:11 mm, 19 mm, SPINE, and 25 mm,这里的11,19,25mm与大家经常应用的尼龙环针座的10,18,24mm相对应的。
产品列表(**19mm和SPINE):
货号 | 产品描述 | 货号 | 产品描述 |
60115-S | MicroMounts, aperture 10µm, pin SPINE | 60115-19 | MicroMounts, aperture 10µm, pin 19mm |
60116-S | MicroMounts, aperture 20µm, pin SPINE | 60116-19 | MicroMounts, aperture 20µm, pin 19mm |
60117-S | MicroMounts, aperture 30µm, pin SPINE | 60117-19 | MicroMounts, aperture 30µm, pin 19mm |
60118-S | MicroMounts, aperture 50µm, pin SPINE | 60118-19 | MicroMounts, aperture 50µm, pin 19mm |
60119-S | MicroMounts, aperture 75µm, pin SPINE | 60119-19 | MicroMounts, aperture 75µm, pin 19mm |
60120-S | MicroMounts, aperture 100µm,pin SPINE | 60120-19 | MicroMounts, aperture 100µm, pin 19mm |
60121-S | MicroMounts, aperture 150µm,pin SPINE | 60121-19 | MicroMounts, aperture 150µm, pin 19mm |
60122-S | MicroMounts, aperture 200µm,pin SPINE | 60122-19 | MicroMounts, aperture 200µm, pin 19mm |
60123-S | MicroMounts, aperture 300µm,pin SPINE | 60123-19 | MicroMounts, aperture 300µm, pin 19mm |
60124-S | MicroMounts, aperture 400µm,pin SPINE | 60124-19 | MicroMounts, aperture 400µm, pin 19mm |
60125-S | MicroMounts, aperture 500µm,pin SPINE | 60125-19 | MicroMounts, aperture 500µm, pin 19mm |
60126-S | MicroMounts, aperture 600µm,pin SPINE | 60126-19 | MicroMounts, aperture 600µm, pin 19mm |
参考资料:
"Microfabricated mounts for high-throughput macromolecular cryocrystallography"
R. E. Thorne, Z. Stum, J. Kmetko, K. O'Neill and R. Gillilan
JOURNAL OF APPLIED CRYSTALLOGRAPHY
36: 1455-1460 December 2003
"A new crystal mounting method for macromolecular crystallography"
Z. Stum, J. Kmetko, K. O'Neill, R. L. Gillilan and R. E. Thorne
SYNCHROTRON RADIATION NEWS
17 (2): 31-32 April 2004