NEW STRATEGIES FOR CONTROLLING CRYSTALLIZATION

Lead Research Organisation: University of Leeds
Department Name: Sch of Chemistry

Abstract

Crystallization is a phenomenon which touches every person, each day of their lives. It lies at the heart of a vast range of fields in science and technology, including pharmaceuticals, healthcare, nanomaterials and foodstuffs, as well as environmental issues like weathering and carbon-capture. Only by building a fundamental understanding of crystal nucleation and growth can we hope to control these processes. Indeed, there has recently been a leap in our understanding of nucleation and growth thanks to advances in analytical techniques, which have enabled study of the nanoscale processes which govern crystallization.

We are addressing this challenge by developing strategies to design and produce crystals with defined polymorph, orientation, morphology and size. Over the last five years we have built expertise and collaborations to study crystallization using techniques such as synchrotron powder XRD, high-resolution cryo-TEM, AFM, EXAFS and the surface force apparatus. This has enabled us to show, for example, that crystallization from vapour often proceeds via liquid condensates in surface pores, which has important implications for ice formation in clouds and climate modelling. We are especially interested in bio-inspired crystallization, where the remarkable materials that are biominerals provide inspiration for the design and formation of synthetic crystals under ambient conditions. Here, we have shown that in contrast to current theories which emphasise control using biomolecules, nature uses physical confinement as a major route to controlling crystallization. We have also used bio-inspired strategies to produce "artificial biominerals" that have mechanical properties equivalent to their biogenic counterparts.

The flexible resources available within this Platform Grant will be used to underpin our existing research portfolio and to explore new ideas and approaches to understanding and controlling crystallization. Attainment of this ambitious goal requires a strategy which includes longer-term, higher-risk research. Particular emphasis will be placed on the use of the physical environment -confinement and surface topography - to control crystallization. As examples, we will extend existing projects to investigate the combined effects of confinement and surface chemistry, and to study the crystallization of amorphous minerals within carbon nanotubes. We will explore new directions including the control of protein crystallization, and microfluidic devices for the study of crystallization within droplets, with the expectation of building new research programmes based on the results.

The Platform Grant will also provide a superior research and training environment for PDRAs and students. Crystallization is a truly interdisciplinary subject, and a particular strength of our group is its ability to take both chemical and physical perspectives on the subject. This grant will hence be particularly valuable in providing a cohesive framework across the Schools of Physics and Chemistry at Leeds to help our researchers to work together as an integrated team. Staff funded by the grant will be assigned to projects rather than individual investigators, thereby enhancing the strategic nature of the platform support. Furthermore, we will provide our PDRAs with enhanced career stability and continuity, and with superior professional development opportunities to help them to apply for competitive lectureships/fellowships or positions in industry. Providing "added value", the Platform grant will be used to initiate/ strengthen collaborations with other internationally leading groups both within and outside the UK, and the PDRAs will gain enormously from research exchanges with other labs. The flexibility of the grant ensures that we retain a critical mass of researchers in key areas (eg microfluidics, AFM) and that our large research group which is funded by many different grants remains integrated, responsive and dynamic.

Planned Impact

This Platform Grant will make an impact on society, people, the economy and the knowledge base by supporting our work on crystallisation. The overarching goal of our research is to generate crystalline materials with target properties, where our strategy is based upon the development of a superior understanding of crystallization phenomena, and how they can be controlled by design. The physical and chemical properties of crystalline materials are defined by features including the polymorph, size, morphology, orientation and purity. As examples, pharmaceuticals are required as single polymorphs, while the properties of quantum dots are intimately linked to their sizes. Analysis of protein structure relies on the generation of large single crystals, while structured hydroxyapatite/ polymer composites are required for bone and dental implants. The catalytic properties of many materials can be optimised according to the exhibition of specific crystal faces, while the processing of powders depends upon their sizes and shapes. All of these can be achieved through effective control over crystal nucleation and growth processes.

At its outset, this Platform Grant will support and develop our work on the use of physical environments - in the form of confinement and topography - to direct crystal nucleation and growth, where this topic has been identified as an area of immediate strategic need for the group. Through this strategy we will lay the groundwork for researchers to use confinement and topography to (1) generate crystals with defined polymorphs, sizes and morphologies, (2) control the location and distribution of crystals on surfaces, (3) define the orientation of crystals and (4) prevent unwanted crystallization. This approach is quite distinct from current methods which invariably rely on the surface chemistry of a substrate or additive, and thus has the potential to be transformative. As the term of the Grant progresses we will diversify the areas supported, and will bring additional resources to our work on additive-controlled crystallisation, and the formation of crystals with composite structures.

Our research therefore has the potential to make a significant impact on the huge number of academic and industrial researchers whose work requires control over crystallisation, on the wider public, and also on the UK economy, in areas ranging from healthcare to nanotechnology to everyday technologies. Links with industry will be made through existing contacts (P&G, Unilever and Nexia Solutions) who are all interested in the control of crystal growth on surfaces, in solution, and in microencapsulation and via the University of Leeds' industry-directed innovation hubs.

The platform grant will also have an important impact on the PDRAs it will support, as well as on our group as a whole in that it will enable new research directions to be explored, new collaborations to be initiated and existing ones to be strengthened and new skills to be acquired. There will be outstanding opportunities for professional development for all members of our group, which will enable them to move smoothly into permanent positions. We will also support the UK in generating researchers with a wide range of valuable generic and scientific skills (such as microfluidics, electron microscopy, XRD). Finally, the public will benefit through enhanced outreach work, in which the PDRAs and the whole group will participate. The visual appeal of crystals will ensure the success of such activities within schools and the wider community. Crystal growth can readily be displayed in real time to an audience with the help of microscopes and computer projection, and the investigators have experience in such outreach activities. The Leeds Centre for Crystallization will be used to offer summer placements in our university to undergraduate students and sixth-form pupils, who will be encouraged to participate via the annual "Leeds Festival of Science".

Publications

10 25 50
 
Description 1. We have been using microfluidic devices to study crystallization on-a-chip, and shown that we can use localised heating to induce transformation of amorphous calcium carbonate (ACC) to single crystals of calcite. It should be possible to extend this strategy to technologically-important materials.

2. We have demonstrated that confinement can be used to promote the aragonite polymorph of calcium carbonate. Calcium carbonate is a widespread compound, whose two common crystalline forms, calcite and aragonite, are important biominerals. Although aragonite is only marginally less stable than calcite under ambient conditions, it usually only crystallizes from solution at high temperatures or in the presence of magnesium ions. Yet organisms readily form both calcite and aragonite biominerals, a capacity usually attributed to the action of specific organic macromolecules. By investigating calcium carbonate precipitation in submicron pores, we have shown that aragonite is promoted in confinement and that pure aragonite crystallizes in nanoscale pores in the absence of any additives. This is of great significance to biomineralization processes, which invariably occur in small volumes, and suggests that organisms may exploit confinement effects to control polymorph.

3. We have initiated new work on the crystallization of proteins on substrates patterned with topographical features, and shown that we can control the nucleation of protein crystals with appropriate topographical features.

4. We have developed a highly-reproducible strategy to occlude gold nanoparticles within single crystals of a range of materials. Acidic macromolecules are considered fundamental to calcium carbonate biomineralisation, and have long been first-choice in the bio-inspired synthesis of crystalline materials. We have obtained data that challenge this view and demonstrate that low-charge macromolecules, which are invariably neglected, can vastly out-perform their acidic counterparts in the synthesis of nanocomposites. This raises important questions about the roles of different classes of proteins in biomineralisation. Using gold nanoparticles functionalised with generic proteins and synthetic homopolymers as additives, we have shown that high concentrations of glycoprotein-functionalised nanoparticles can be incorporated within calcite single crystals, while hydroxylated polymers were even more effective, yielding dark red crystals containing 37 wt% nanoparticles. The nanoparticles were perfectly dispersed within the host crystal and so close together that they exhibited plasmon coupling. The versatility of this strategy is also demonstrated by extension to alternative host crystals. This simple and scalable occlusion approach opens the door to a novel class of single crystal nanocomposites.
5. We have developed the use of X-ray tomography and X-ray diffraction tomography to study crystallisation in situ within nanoporous media. Crystallisation within porous media is an important phenomenon in the environment, biology and materials synthesis, but remains poorly understood. Synchrotron X-ray computed tomography and diffraction tomography were used to reveal how a population of calcium sulfate particles develop over time within nanoporous glass rods. The exceptional image resolution achievable enabled us to characterise the crystallisation pathway and demonstrate that this is governed by the local surface chemistry. We have shown that crystallisation is hugely retarded in these environments, and that following the precipitation of an amorphous precursor phase, gypsum (CaSO4.2H2O) can precipitate either directly or via metastable bassanite (CaSO4.0.5H2O). This demonstration of multiple pathways potentially unites many conflicting reports regarding gypsum precipitation. Conversely, while crystallisation within the rod causes no structural damage, gypsum precipitation adjacent to the rod surface causes catastrophic fracture. Understanding crystallisation in porous media ultimately promises the ability to control processes including weathering, scaling, contaminant sequestration and inorganic biomaterials synthesis.

6. We have developed the capability to study crystallisation within flowing droplets in a microfluidic device using synchrotron X-ray diffraction, and used this to study crystallisation mechanisms. Droplet Microfluidics-Coupled X-ray Diffraction (DMC-XRD) enables the collection of time-resolved, serial diffraction patterns from a stream of flowing droplets containing growing crystals. The droplets offer reproducible reaction environments, and radiation damage is effectively eliminated by the short residence time of each droplet in the beam. We have then used DMC-XRD to identify effective particulate nucleating agents for calcium carbonate and to study their influence on the crystallization pathway. Bioactive glasses and NX illite were shown to significantly lower the induction time, highlighting the importance of both surface chemistry and topography on the nucleating efficiency of a surface. This technology is also extremely versatile, and could be used to study dynamic reactions with a wide range of synchrotron-based techniques.

7. We have used the natural "pockets" present on cleaved mica to investigate how surface topography can direct ice nucleation. Crystal nucleation - the first appearance of a crystalline phase where there was none before - usually occurs at the surface of a foreign material. Ice formation in the atmosphere is dependent upon the number and type of aerosol particles present, but little is known about why some are more effective than others. We have investigated the role of surface topography in promoting crystallisation of ice and different organic crystals and shown that acute geometries are highly effective in promoting the growth of a confined crystalline phase, which then gives rise to a bulk phase. This is relevant to crystallisation in a large number of real world systems such as industrial film growth and our climate.

8. Understanding how surfaces direct nucleation is a complex problem that limits our ability to predict and control crystal formation. We have addressed this challenge using high-speed imaging to identify and quantify the sites at which ice nucleates in water droplets on macroscopic feldspar substrates. Our data demonstrate that ice nucleation only occurs at a tiny number of locations, all of which are associated with micron-size pits. Similar behaviour is observed on a-quartz substrates that lack cleavage planes. These results demonstrate that substrate heterogeneities are the salient factor in promoting nucleation, and therefore finally prove the existence of active sites. We have also obtained strong evidence that the activity of these sites derives from a combination of surface chemistry and nanoscale topography. These results have implications for the nucleation of a wide range of materials and suggest new strategies for promoting or inhibiting nucleation across a wide range of applications.

9. We have developed a microfluidic-based method to understand crystallisation mechanisms. As crystallization processes are often occur rapid, it can be difficult to monitor the growth mechanisms. We have used the fact that crystallization proceeds more slowly in small volumes than in bulk solution, to investigate the effects of the soluble additives Mg2+ and poly(styrene sulfonate) on the early stages of growth of calcite crystals. Using a "Crystal Hotel" microfluidic device to provide well-defined, nanoliter volumes, we showed that calcite crystals form via an amorphous precursor phase. Surprisingly, the first calcite crystals formed were perfect rhombohedra, and no morphological influence of the soluble additives were observed until the crystals reach sizes of 0.1-0.5 µm for Mg2+ and 1-2 µm for PSS. The crystals then continued to grow to develop morphologies characteristic of these additives. These results can be rationalized by considering additive binding to kink sites, which is consistent with crystal growth via a classical mechanism.

10. We have used confinement to study the crystallisation of amorphous calcium carbonate (ACC). Using a crossed-cylinder apparatus, we have directly observed that ACC forms in confinement in very dilute systems where it is virtually undetectable in bulk solution. The aggregation of ACC particles is also seen to significantly decrease with increasing confinement, suggesting that the stability of ACC in confinement is related to its aggregation state and restricted mobility. Finally, we also observed that ACC transforms to calcite via the metastable phase vaterite in constrained volumes at solution concentrations where direct transformation to calcite occurs in bulk solution. This work underscores the utility of using confinement to detect and preserve transient metastable phases in bulk solution, where this may have applications in materials syntheses utilising amorphous precursor phases.

11. Super resolution microscopy is predominantly used to study biological samples. We have applied this technique to the analysis of an inorganic sample - a nanocomposite comprising nanoparticles embedded within a single crystal host. To fully profit from these materials it is essential to be able to characterise their 3D structures, identifying the locations of individual nanoparticles, and the defects present within the host crystals. Using calcite crystals containing quantum dots as a model system, we used 3D stochastic optical reconstruction microscopy (STORM) to locate the positions of the nanoparticles within the host crystal. The nanoparticles are shown to preferentially associate with dislocations in a manner previously recognised for atomic impurities, rendering these defects visible by STORM. Our images also demonstrate that the types of dislocations formed at the crystal/ substrate interface vary according to the nucleation face, and dislocation loops are observed that have entirely different geometries to classic misfit dislocations. This approach offers a rapid, easily accessed, and non-destructive method for visualising the dislocations present within crystals, and gives insight into the mechanisms by which additives become occluded within crystals.

12. Understanding how surfaces direct nucleation is a complex problem that limits our ability to predict and control crystal formation. We have addressed this challenge using high-speed imaging to identify and quantify the sites at which ice nucleates in water droplets on the two natural cleavage faces of macroscopic feldspar substrates. Our data show that ice nucleation only occurs at a few locations, all of which are associated with micron-size surface pits. Similar behaviour is observed on a-quartz substrates that lack cleavage planes. These results demonstrate that substrate heterogeneities are the salient factor in promoting nucleation, and therefore finally prove the existence of active sites. We also provide strong evidence that the activity of these sites derives from a combination of surface chemistry and nanoscale topography. Our results have implications for the nucleation of many materials and suggest new strategies for promoting or inhibiting nucleation across a wide range of applications.

13. Understanding atmospheric ice nucleation is a key component to building an understanding of weather and climate. Nucleation is largely determined by the presence of rare "active sites" present on air-borne particles. These sites may promote nucleation by one of two distinct pathways: by immersion of the site in supercooled liquid water or by its exposure to supersaturated water vapour. Our work compares these two pathways by identifying active nucleation sites on thin sections of two atmospherically important minerals (feldspar and quartz) both from vapour and from liquid water. We find that the two sets of sites have little correlation, with only 6 out of 73 sites active for nucleation from liquid also active for nucleation from vapour. This implies different chemical and topographical requirements for nucleation sites for each pathway, and that the activity of a site can only be considered within the context of the specific nucleation pathway involved.
Exploitation Route Our work will enable researchers/ scientists to use confinement to control crystallisation, where this in important to the enormous range of processes that occur in confinement. This include the formation of biomaterials and biominerals, processes such as weathering and frost-heave and the templating of nanomaterials.

Polymorph control is one of the most fascinating topics in crystallisation - and one which is still poorly understood. That confinement can control polymorph - and indeed select calcite or aragonite - gives us new understanding of the factors which govern polymorph, and can potentially answer the long-standing "calcite/aragonite" problem.

There is huge need to control protein crystallisation to generate XRD-quality crystals for structural analysis. That surface topography can potentially be used to control protein crystallisation is therefore of interest in all working in this area.

Our development of expertise to use microfluidic devices to perform synchrotron X-ray analysis of continuous and segmented-flow systems is expected to have a significant impact on researchers from a wide range of backgrounds.
Sectors Chemicals,Construction,Environment,Healthcare,Culture, Heritage, Museums and Collections,Pharmaceuticals and Medical Biotechnology

URL http://www1.chem.leeds.ac.uk/FCM/
 
Description Flow-Xl: A New UK Facility for Analysis of Crystallisation in Flow Systems
Amount £1,129,048 (GBP)
Funding ID EP/T006331/1 
Organisation Engineering and Physical Sciences Research Council (EPSRC) 
Sector Public
Country United Kingdom
Start 01/2020 
End 01/2022
 
Description Programme Grant
Amount £5,436,236 (GBP)
Funding ID EP/R018820/1 
Organisation Engineering and Physical Sciences Research Council (EPSRC) 
Sector Public
Country United Kingdom
Start 03/2018 
End 02/2023
 
Title Dataset for 'An Innovative Data Processing Method for Studying Nanoparticle Formation in Droplet Microfluidics using X-rays Scattering' 
Description X-ray scattering techniques provide a powerful means of characterizing the formation of nanoparticles in solution. Coupling these techniques to segmented-flow microfluidic devices that offer well-defined environments gives access to in situ time-resolved analysis, excellent reproducibility, and eliminates potential radiation damage. However, analysis of the resulting datasets can be extremely time-consuming, where these comprise frames corresponding to the droplets alone, the continuous phase alone, and to both at their interface. We here describe a robust, low-cost, and versatile droplet microfluidics device and use it to study the formation of magnetite nanoparticles with simultaneous synchrotron SAXS and WAXS. Lateral outlet capillaries facilitate the X-ray analysis and reaction times of between a few seconds and minutes can be accommodated. A two-step data processing method is then described that exploits the unique WAXS signatures of the droplets, continuous phase, and interfacial region to identify the frames corresponding to the droplets. These are then sorted, and the background scattering is subtracted using an automated frame-by-frame approach, allowing the signal from the nanoparticles to be isolated from the raw data. Modeling these data gives quantitative information about the evolution of the sizes and structures of the nanoparticles, in agreement with TEM observations. This versatile platform can be readily employed to study a wide range of dynamic processes in heterogeneous systems. 
Type Of Material Database/Collection of data 
Year Produced 2021 
Provided To Others? Yes  
URL https://archive.researchdata.leeds.ac.uk/977/
 
Title High-speed Imaging of Ice Nucleation in Water Proves the Existence of Active Sites - dataset 
Description This is the data for High-speed Imaging of Ice Nucleation in Water Proves the Existence of Active Sites, where we demonstrate that specific locations on the surface of feldspar and quartz are repeatedly the area in which ice growth originates in freezing experiments, i.e. the nucleation active sites. This means that heterogeneities on the surfaces are of first order importance in determining the temperature at which liquid water freezes. The data contained in this repository consists of the ice nucleation experimental data, used to construct the figures in the main text and supplementary materials. These show that nucleation is site specific for both quartz and feldspar. We also present WDS mapping results, where no chemical differences were found at the active sites within the resolution of the technique, leading to our hypothesis that topography is an important factor in determining ice nucleation effectiveness of a site. 
Type Of Material Database/Collection of data 
Year Produced 2019 
Provided To Others? Yes  
 
Title The effect of additives on the early stages of growth of calcite single crystals - dataset 
Description As crystallization processes are often rapid, it can be difficult to monitor the growth mechanisms. In this study, we 
Type Of Material Database/Collection of data 
Year Produced 2018 
Provided To Others? Yes  
 
Description Nik Kapur microfluidics 
Organisation University of Leeds
Department Institute of Transport Studies
Country United Kingdom 
Sector Academic/University 
PI Contribution Our research team has identified the problem that needs to be solved - characterisation of inorganic materials by XRD on-chip. We contribute expertise in materials chemistry and XRD analysis.
Collaborator Contribution Nik Kapur has assisted in the design of microfluidic devices that can be used for synchrotron XRD analysis of crystallisation within droplets. His expertise in fluid dynamics and the design and manufacture of devices has been invaluable.
Impact 1 paper to date. Multi-disciplinary collaboration between chemistry and engineering.
Start Year 2014
 
Description Prof Chick Wilson 
Organisation University of Bath
Department School of Health Bath
Country United Kingdom 
Sector Academic/University 
PI Contribution We helped them carry out in situ XRD analysis of crystallisation in millifluidic segmented flow reactors at Diamond light source and analyse their data
Collaborator Contribution They participated in a number of joint beam-times
Impact We are currently writing a number of joint publications and grant proposal
Start Year 2016
 
Description Prof Naomi Chayen 
Organisation Imperial College London
Department Faculty of Medicine
Country United Kingdom 
Sector Academic/University 
PI Contribution We have collaborated on using out model confined systems to control and study protein crystallisation
Collaborator Contribution Naomi hosted a postdoc to teach them methods to crystallise proteins. We have developed ideas together.
Impact Publications and a grant proposal are in preparation
Start Year 2016
 
Description Prof Nico Sommerdijk 
Organisation Eindhoven University of Technology
Country Netherlands 
Sector Academic/University 
PI Contribution We have collaborated on a project investigating the role of confinement in the mineralisation of collagen
Collaborator Contribution They contributed expensive TEM time
Impact We have recently submitted a paper on this work
Start Year 2010
 
Description Bristol Festival of Nature 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Public/other audiences
Results and Impact 6 members of the research group participated in the 2-day "Bristol festival of nature" where they manned a stand that showcased demonstrations about crystallisation processes.
Year(s) Of Engagement Activity 2015
 
Description Exhibition of Images 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Public/other audiences
Results and Impact 6 week exhibition of images of crystals at the North Bar Leeds
Year(s) Of Engagement Activity 2017
 
Description Exhibition of Scientific Images 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Public/other audiences
Results and Impact 6 week exhibition of images of crystals at North bar leeds
Year(s) Of Engagement Activity 2016
 
Description Exhibition of scientific images 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Public/other audiences
Results and Impact Exhibition of images of crystals at the North Bar, Leeds
Year(s) Of Engagement Activity 2019
 
Description Lancaster Science Festival 2016 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Public/other audiences
Results and Impact 7 members of the research group participated in the 2-day "Lancaster Science festival" where they manned a stand that showcased demonstrations about crystallisation processes.
Year(s) Of Engagement Activity 2015