Crystal Growth 101
The complete collection of eighteen Crystal Growth 101 articles. Introduction, Sample Preparation for Crystallization, Crystallization Screening, Sitting Drop Vapor Diffusion, Hanging Drop Vapor Diffusion, Microbatch Crystallization, Microdialysis Crystallization, Viewing Crystallization Experiments, Salt or Protein Crystals, Optimization, Drop Ratio, Temperature as a Crystallization Variable, Seeding, Buffer Formulation, Phosphate Buffer Dilution Table, Using Volatile Buffers to Adjust Drop pH and Induce Crystallization, Reagent Formulation and Handling, and Halides and Phasing.
18 篇晶体生长 101 篇文章的完整合集。 简介、结晶样品制备、结晶筛选、坐滴蒸气扩散、悬滴蒸气扩散、微批次结晶、微透析结晶、观察结晶实验、盐或蛋白质晶体、优化、滴比、温度作为结晶变量、晶种、缓冲液配方 , 磷酸盐缓冲液稀释表，使用挥发性缓冲液调节液滴 pH 值和诱导结晶、试剂配制和处理以及卤化物和定相。
Crystal Growth 101 Introduction 晶体生长101简介
The Crystal Growth 101 series prepared by Hampton Research presents an overview of the preparation of the sample, methods, screening, optimization, reagent formulation, and other aspects of protein crystallization. We hope Crystal Growth 101 will prove a useful resource and inspiration in your crystal quest. Best of success with your crystals!
Hampton Research 编写的 Crystal Growth 101 系列概述了蛋白质结晶的样品制备、方法、筛选、优化、试剂配方和其他方面。 我们希望 Crystal Growth 101 将成为您水晶探索中有用的资源和灵感。 用你的水晶取得最大的成功！
结晶样品制备 Sample Preparation for Crystallization
The sample is the single most important variable in the crystallization experiment. Begin with a pure, homogeneous, stable, active sample. The sample should be as pure as possible, 95 to 98%, assayed by Coomassie stained SDS-PAGE. A homogeneous, active sample, free of contaminants, aggregates, and minimal conformational flexibility is desired.
样品是结晶实验中最重要的单一变量。从一个纯的，均匀的，稳定的，活跃的样品开始。样品应尽可能纯净，95%~98%，用考马西染色SDS-PAGE法测定。一个均匀的，活跃的样品，没有污染物，聚集体，和最小的构象灵活性是需要的。
Crystal Growth 101 Screening 晶体生长 101 筛选
Crystallization screening is the proce
ss of evaluating methods, reagents, and other chemical and physical variables with the objective of producing crystals and/or identifying the variables which are positively or negatively associated with crystallization of the sample.
At the time of this writing, up to 40% of samples screened for crystallization will produce some kind of crystalline result and 10% of samples will produce a crystal suitable for X-ray diffraction analysis. About 75% of proteins screened require optimization. Optimization is the systematic manipulation and evaluation of variables which influence the crystallization of the sample.
结晶筛选是评估方法、试剂和其他化学和物理变量的过程，目的是生产晶体和/或识别与样品结晶正相关或负相关的变量。
在撰写本文时，多达 40% 的筛选结晶样品会产生某种结晶结果，10% 的样品会产生适合 X 射线衍射分析的晶体。 大约 75% 的筛选蛋白质需要优化。 优化是对影响样品结晶的变量的系统操作和评估。
Crystal Growth 101 Sitting Drop Vapor Diffusion 晶体生长 101 坐滴蒸气扩散
The complete collection of eighteen Crystal Growth 101 articles. Introduction, Sample Preparation for Crystallization, Crystallization Screening, Sitting Drop Vapor Diffusion, Hanging Drop Vapor Diffusion, Microbatch Crystallization, Microdialysis Crystallization, Viewing Crystallization Experiments, Salt or Protein Crystals, Optimization, Drop Ratio, Temperature as a Crystallization Variable, Seeding, Buffer Formulation, Phosphate Buffer Dilution Table, Using Volatile Buffers to Adjust Drop pH and Induce Crystallization, Reagent Formulation and Handling, and Halides and Phasing.
The sitting drop vapor diffusion technique is another popular method for the crystallization of macromolecules. The principle of vapor diffusion is straightforward. A drop composed of a mixture of sample and reagent is placed in vapor equilibration with a liquid reservoir of reagent. Instructional video for Sitting Drop Vapor Diffusion
18 篇晶体生长 101 篇文章的完整合集。 简介、结晶样品制备、结晶筛选、坐滴蒸气扩散、悬滴蒸气扩散、微批次结晶、微透析结晶、观察结晶实验、盐或蛋白质晶体、优化、滴比、温度作为结晶变量、晶种、缓冲液配方 , 磷酸盐缓冲液稀释表，使用挥发性缓冲液调节液滴 pH 值和诱导结晶、试剂配制和处理以及卤化物和定相。
坐滴蒸汽扩散技术是另一种流行的大分子结晶方法。 蒸汽扩散的原理很简单。 一滴由样品和试剂的混合物组成的液滴与试剂的液体容器处于蒸汽平衡状态。 坐滴蒸气扩散的教学视频
Crystal Growth 101 Hanging Drop Vapor Diffusion 晶体生长 101 悬滴蒸气扩散
The hanging drop vapor diffusion technique is a popular method for the crystallization of macromolecules. The principle of vapor diffusion is straightforward. A drop composed of a mixture of sample and reagent is placed in vapor equilibration with a liquid reservoir of reagent. Typically the drop contains a lower reagent concentration than the reservoir. To achieve equilibrium, water vapor leaves the drop and eventually ends up in the reservoir. As water leaves the drop, the sample undergoes an increase in relative supersaturation. Both the sample and reagent increase in concentration as water leaves the drop for the reservoir. Equilibration is reached when the reagent concentration in the drop is approximately the same as that in the reservoir. Instructional Video for Hanging Drop Vapor Diffusion
悬滴蒸气扩散技术是一种流行的大分子结晶方法。 蒸汽扩散的原理很简单。 一滴由样品和试剂的混合物组成的液滴与试剂的液体容器处于蒸汽平衡状态。 通常，液滴包含比容器低的试剂浓度。 为了达到平衡，水蒸气离开液滴并最终进入水库。 随着水离开液滴，样品的相对过饱和度增加。 当水离开水滴进入水库时，样品和试剂的浓度都会增加。 当液滴中的试剂浓度与储液器中的浓度大致相同时，即达到平衡。 悬滴蒸气扩散教学视频
Crystal Growth 101 Microbatch Crystallization 晶体生长101】
Microbatch crystallization is a method where the sample and reagent are combined and sealed in a plate, tube, container, or sealed under a layer of oil.
Oils can also be used as a barrier between the reservoir and the drop in traditional Hanging or Sitting Drop crystallization experiments. This is known as Vapor Diffusion Rate Control.
微批次结晶是一种将样品和试剂混合并密封在板、管、容器中或密封在一层油下的方法。
在传统的悬滴或坐滴结晶实验中，油也可以用作油藏和液滴之间的屏障。 这称为蒸汽扩散速率控制。
Converting Vapor Diffusion to Microbatch 将蒸汽扩散转化为微批次
When the conditions for growing protein crystals by sitting or hanging drop vapor diffusion are known, crystals of comparable or sometimes better quality can usually be produced by microbatch under oil using a general set of guidelines.
当通过静置或悬滴蒸汽扩散生长蛋白质晶体的条件已知时，通常可以使用一组通用指南在油下通过微批次生产具有可比性或有时更好质量的晶体。
Vapor Diffusion to Batch Calculator 蒸汽扩散到批次计算器
Vapor Diffusion to Batch Calculator is an Excel based spreadsheet that allows one to enter vapor diffusion crystallization condition data in order to generate a best guess microbatch condition. Vapor diffusion crystallization condition data is entered into the yellow fields and best guess microbatch conditions, results, and calculations are displayed in the green fields.
Vapor Diffusion to Batch Calculator 是一种基于 Excel 的电子表格，允许输入蒸气扩散结晶条件数据以生成最佳猜测微批次条件。 蒸汽扩散结晶条件数据输入黄色字段，最佳猜测微批次条件、结果和计算显示在绿色字段中。
Crystal Growth 101 Microdialysis Crystallization 晶体生长101微渗析结晶
In the microdialysis crystallization method the sample is separated from the crystallization reagent by a semi-permeable membrane. The semi-permeable membrane allows small molecules, such as salts, additives, and other crystallization reagents to pass, but prevents biological macromolecules from crossing the membrane. Crystallization of the sample takes place due to the diffusion of crystallization reagent out of, or into the sample, at constant sample concentration.
在微透析结晶法中，样品通过半透膜与结晶试剂分离。 半透膜允许小分子，如盐、添加剂和其他结晶试剂通过，但阻止生物大分子穿过膜。 由于在恒定的样品浓度下结晶试剂从样品中扩散出来或扩散到样品中，从而发生样品的结晶。
Crystal Growth 101 Viewing Crystallization Experiments 查看结晶实验
A stereomicroscope with zoom (8 to 12.5:1) and 10 to 100x magnification, LED illuminators to minimize temperature change during observation, polarizing optics to discern birefringence, camera attachment, and a viewing platform large enough to support a crystallization plate when viewed from any and all wells is a good starting point for observing the experiment. Automated imaging systems are also available, as well as imaging systems with ultraviolet and other capabilities.
晶体生长 101 查看结晶实验
具有变焦（8 至 12.5:1）和 10 至 100 倍放大倍率的立体显微镜、可将观察过程中的温度变化降至最低的 LED 照明器、可识别双折射的偏振光学器件、相机附件以及足够大的观察平台，可在从任何角度观察时支撑结晶板 并且所有孔都是观察实验的良好起点。 还提供自动成像系统，以及具有紫外线和其他功能的成像系统。
Crystal Growth 101 Salt or Protein Crystals? 盐或蛋白质晶体？
In viewing crystallization experiments, when crystals are observed, one must determine if the crystals are of the target biological macromolecule (protein) or salt (inorganic, small molecule) crystals. Before deciding on how to distinguish whether the crystal is salt or protein,take a picture of the crystal for documentation before you destroy the crystal or make it disappear.
在观察结晶实验中，当观察到晶体时，必须确定晶体是目标生物大分子（蛋白质）还是盐（无机、小分子）晶体。 在决定如何区分晶体是盐还是蛋白质之前，在破坏晶体或使其消失之前，先拍下晶体的照片以备文档。
Crystal Growth 101 Optimization 晶体生长 101 优化
Optimization is the manipulation and evaluation of biochemical, chemical, and physical crystallization variables, towards producing a crystal, or crystals with the specific desired characteristics.
优化是对生化、化学和物理结晶变量的操作和评估，以生产具有特定所需特征的晶体或晶体。
Crystal Growth 101 Drop Ratio 晶体生长101掉落率
Varying the drop ratio is a simple method to utilize during crystallization screening and optimization. Drop ratios allow one to explore varying levels of initial and final protein and reagent concentration, explore different equilibration paths, and cover a wider range of relative supersaturation. All by simply changing the amount of sample and reagent added to the drop.
在结晶筛选和优化过程中，改变下降率是一种简单的方法。 下降率允许人们探索不同水平的初始和最终蛋白质和试剂浓度，探索不同的平衡路径，并涵盖更广泛的相对过饱和度。 只需更改添加到液滴中的样品和试剂的量即可。
Drop Ratio Calculator 下降率计算器
Drop Ratio Calculator is an Excel based spreadsheet that will display the effects of varying the drop ratio of sample and crystallization reagent in a vapor diffusion experiment. Protein Concentration and Reagent concentration can be entered into the gray fields and the results displayed above in the gray fields. Custom Protein Concentration and Custom Reagent Concentration can be entered into the blue fields, along with Custom Protein Volume and Custom Reagent Volume and the results displayed above in the blue fields.
Drop Ratio Calculator 是一个基于 Excel 的电子表格，它将显示在蒸汽扩散实验中改变样品和结晶试剂的下降率的影响。 蛋白质浓度和试剂浓度可以输入灰色字段，结果显示在灰色字段中。 Custom Protein Concentration 和 Custom Reagent Concentration 可以输入到蓝色字段中，以及 Custom Protein Volume 和 Custom Reagent Volume 以及上面蓝色字段中显示的结果。
Crystal Growth 101 Temperature 晶体生长 101 温度
Temperature can be a significant variable in the crystallization of biological macromolecules (proteins). Temperature often influences nucleation and crystal growth by manipulating the solubility and supersaturation of the sample. Temperature has also been shown to be an important variable with phase separation in detergent solutions during membrane protein crystallization.
温度可能是生物大分子（蛋白质）结晶过程中的一个重要变量。 温度通常通过控制样品的溶解度和过饱和度来影响成核和晶体生长。 温度也已被证明是膜蛋白结晶过程中洗涤剂溶液中相分离的重要变量。
Crystal Growth 101 Seeding 晶体生长 101 播种
Seeding allows one to grow crystals in the Metastable Zone, where spontaneous homogeneous nucleation cannot occur, but crystal growth from seeds can occur. Why would one want to do this? For control, reproducibility, and to improve the likelihood of a successful crystallization experiment. In the Metastable Zone crystals can grow from seeds but cannot spontaneously nucleate. By placing a seed or solution of seeds in a drop which is saturated to the Metastable Zone one can use the seeds to grow larger single crystals. By controlling the number of seeds introduced into the Metastable Zone drop one can control the number of crystals grown. It is not practically possible to measure and know the number of seeds introduced to a drop, but by performing serial dilutions from a concentrated seed stock one can control the number of crystals grown in the Metastable Drop.
晶种允许人们在亚稳区中生长晶体，在亚稳区不能发生自发的均匀成核，但可以发生晶种的晶体生长。 为什么要这样做？ 用于控制、重现性和提高结晶实验成功的可能性。 在亚稳区晶体可以从种子生长但不能自发成核。 通过将种子或种子溶液放入饱和到亚稳区的液滴中，可以使用种子生长更大的单晶。 通过控制引入亚稳区的种子数量，可以控制晶体生长的数量。 测量和知道引入液滴的种子数量实际上是不可能的，但通过从浓缩的种子库中进行连续稀释，可以控制在亚稳态液滴中生长的晶体数量。
Crystal Growth 101 Buffer Table晶体生长 101 缓冲表
Crystallization buffer table in order of useful pH range.
结晶缓冲表按有用的 pH 范围排序。
Crystal Growth 101 Buffer Formulation 晶体生长 101 缓冲液配方
Details about the formulation of Hampton Research buffers, including how to titrate, useful pH range, pKa and more.
有关 Hampton Research 缓冲液配方的详细信息，包括如何滴定、有用的 pH 范围、pKa 等。
Crystal Growth 101 Phosphate Buffer Dilution Table Crystal Growth 101 磷酸盐缓冲液稀释表
Use this table to easily create Sodium potassium phosphate buffers and reagents between 0.2 and 4.0 M while varying the pH between 5 and 8.2.
使用此表可轻松创建 0.2 到 4.0 M 之间的磷酸钠钾缓冲液和试剂，同时在 5 到 8.2 之间改变 pH 值。
Crystal Growth 101 Volatile Buffers 晶体生长 101 挥发性缓冲液
Volatile buffers, when added only to the reagent reservoir of a vapor diffusion experiment, can alter the pH of the crystallization drop by vapor diffusion of the volatile acid or base component from reservoir into the drop. This may be particularly useful when the sample is known to have pH dependent solubility and may be used to induce crystallization.
挥发性缓冲液，当仅添加到蒸气扩散实验的试剂容器时，可以通过挥发性酸或碱成分从容器到液滴的蒸气扩散来改变结晶液滴的 pH 值。 当已知样品具有依赖于 pH 值的溶解度并且可用于诱导结晶时，这可能特别有用。
Crystal Growth 101 Reagent Formulation Crystal Growth 101 试剂配方
Good crystallization results depend on being able to replicate experiments, and experiments cannot be replicated unless the reagents are made correctly and consistently. Be consistent with sources, methods, and techniques. Maintain accurate and detailed records; your future self and perhaps others, will thank you later.
良好的结晶结果取决于能够重复实验，除非正确且一致地制作试剂，否则实验无法重复。 与来源、方法和技术保持一致。 保持准确和详细的记录； 你未来的自己，也许其他人，稍后会感谢你。
Crystal Growth 101 Solubility Table 晶体生长 101 溶解度表
Solubility data for formulating a saturated solution of the salts listed at the designated temperature.
在指定温度下配制所列盐的饱和溶液的溶解度数据。
Crystal Growth 101 Using Halides for Phasing 使用卤化物进行定相的晶体生长 101
Bromide and iodide can diffuse into protein crystals when soaked with the appropriate solution and can successfully be used for phasing.
当用适当的溶液浸泡时，溴化物和碘化物可以扩散到蛋白质晶体中，并可以成功地用于定相。
Crystallization Scoring Sheet 结晶评分表
24 well plate format scoring sheet for screening and optimization.
用于筛选和优化的 24 孔板格式评分表。
Polyethylene Glycol Stability 聚乙二醇稳定性
PEG Stability: A Look at pH and Conductivity Changes over Time in Polyethylene Glycols
PEG 稳定性：观察聚乙二醇中 pH 值和电导率随时间的变化
Hampton Research Catalog
Hampton Research Catalog and Crystal Growth 101 – Complete