Ease of scalability

Producing COVID-19 vaccines for the entire world population rapidly demonstrated the microfluidic development scale up roadblock. Discovery formulations developed using microfluidics had to be scaled up using alternative higher throughput formulating technology with different operating parameters.   

With a growing demand for more mRNA-LNP based vaccines, proven to have high efficacy, there is currently a need to facilitate a faster and cheaper global deployment of a high-throughput manufacturing method. A GMP compliant approach is needed, from the earliest possible stage through to manufacturing, that is also tuneable in size to access various tissues or specific drug targets.  

Advanced cross flow mixing (AXF) from Micropore Technologies allows for seamless scaleup with consistent physics, mechanisms conditions and geometry across its equipment range. Ultimately Micropores AXF technology can be scaled up to a device with over 10 million pores with the potential for a throughput of up to 20 Liters per minute all this from a device that would still fit inside a briefcase. AXF allows controlled, low shear precision continuous flow mixing technology from nano to micro formulations for when you want to avoid roadblocks on your product development journey. It uses the same same shear, same physics and same technology from lab bench to manufacturing scale to enable scale up with confidence.  

Micropore technology address key concerns for adoption

• Expertise, availability of skilled personnel, associated risks, and the need for a reliable supply chain. 

Micropore is a technology provider with global experience in manufacturing all different types of vaccine modalities can further ensure a cost-effective, high-quality process. Partnering with Micropore will enable a stronger benchmark with in-depth expertise and the ability to leverage novel technologies will also help reduce risk and shorten timelines.

• There is a need to develop standard analytical methods for quality characterisation and reference material that lend insight into the mechanisms of stability or degradation of mRNA and LNPs containing it. 

The determinants of stability of mRNA in LNP formulations – what parts are predicated on the payload of mRNA and what portions are predicated on the lipid nanoparticles themselves or what portions are predicated on the freeze-drying cycle that’s used – can be related to its size and its secondary structure. Messenger RNA poses a unique manufacturing challenge because of its large size. Other RNA entities such as siRNA and guide RNA for clustered regularly interspaced short palindromic repeats (CRISPR) technology typically are produced using chemical synthesis, which can be performed in a relatively controlled environment. But mRNAs are larger, with complex three dimensional structures that aren’t yet fully understood. Malvern Zetasizer (DLS) enables particle size and stability measurements while the RedShiftBio Aurora (MMS) system enables secondary structure (HOS) determination.

• The need to move to larger scale manufacturing of mRNA-LNPs without an extensive setup or excess capacity to convert.

With mRNA vaccine production requiring relatively less space than other approaches, new facilities may be more feasible and affordable. Micropore technology can enable localised production of vaccines and thus accelerate access to a much larger population. In locations with limited or no infrastructure, the Micropore approach can be the shortest route to production and can reflect the exact needs of the organisation at minimal cost.

• The need for future vaccine manufacturing facilities to be closed and continuous processing, plus equipment connectivity and communication.

Micropore offers a minimal cost model achieved through the AXF advanced cross flow technology platform. The flexibility of mRNA-based vaccines when manufactured using this single piece of stainless steel equipment with no consumables means it requires the least capital investment. The scalability of production (from 200 μL to 1500 L/hr) reduces the facility design complexity and means that more doses can be manufactured in a continuous process which eliminates variability, compared to a batch-to-batch approach. As such, this vaccine modality combined with the Micropore mixing platform can be a robust starting point for production with low risk.

• The need to overcome current barriers of manufacturing process to be fully digitalised as regulatory authorities rely on data and parameters recorded during production for verification and approval.

Micropore cross flow technology demonstrates predicable scalability which is favourable especially for GMP manufacturing which means process controls can be introduced that are automated – process analytical technologies (PAT). Automated analysis of properties such as online particle size enables the option to automatically control any deviations in size and feed that back to control the pumps and optimise control to give the correct size again. This increases confidence in quality of production meaning throughput can be increased further.

Continuous Formation & Stabilisation of LNPs Minimises Risk of mRNA Degradation

Batch mode vs continuous mode manufacturing: While Impingement jet mixing (IJM) or T-mixers are the most widespread manufacturing method currently, high turbulent mixing combined with high pressure and shear stress effects can compromise LNP stability and affect overall performance of the product. The process also suffers from high batch variability and high wastage due to holding or prolonged processing steps.

Micropore Technologies offers Continuous Manufacturing: Micropore Technologies employs laminar flow mixing across a permanent stainless-steel membrane to produce reproducible, scalable LNPs. The outer dispersed phase is continuously mixed with the aqueous inner compartment to form LNPs. Size controlled uniform particles are generated in a continuous flow capacity of up to 1500 L/hour making this by far the fastest production rate in the LNP industry. This would translate to roughly 58,000 doses of vaccine every minute, an important capability when faced with the demands of global disease emergencies. 

Micropore Technologies: 2^nd generation Vaccine manufacturing Process

Micropore’s Membrane Technology: How it works

Micropore’s equipment is uniquely suited to the production of complex nanomedicines in a scalable manner – from small 200µl samples through to 1 billion doses.

Micropore technologies differs from conventional mixing LNP techniques by considering the process not as discrete or separate unit operations but as one single whole process.

Attributes:

• High efficiency, scalable and reproducible.

• Reduced manufacturing time and costs.

• Customisation for diverse LNP formulations.

• Tunable particle size with narrow size distribution (PDI).

The Micropore system comprises a membrane inside a housing. Lipid mixtures added from the top are mixed with the RNA/buffer solution from the side which enters the membrane to produce monodisperse particles.  The system operates at very low pressure – approximately 2 Bar. This allows the size of the particles created to be very predictable with any change in flow rate. T-mixer devices operate at much higher pressures – about 30 Bar.

The graphs above show comparison control curves for AXF vs. T-mixer.

Advanced Cross Flow (AXF) mixing

Micropore’s equipment is uniquely suited to the production of complex nanomedicines in a scalable manner – from mL batches up to tonnes.

A lipid/organic phase passes through the 100,000s of membrane pores into the flow of aqueous continuous phase that passes through the centre of the membrane tube. Gentle, laminar flow based mixing allows for good preservation of sensitive materials. Precision engineered equipment gives great size distributions even at scale, allowing precise targeting of distributions.

Process robustness and high encapsulation efficiency

Here we present data from a formulation prepared using the AXF Pathfinder 20.

The conditions used are listed below:

• Formulation ratio DDAB:DSPC:cholesterol:DMS-PEG2000, 40:10:48:2 mol%.
• Lipid concentration, 3.5mg/ml
• N:P ratio, 1:6
• polyA concentration, 0.046mg/ml
• Flow rate ratio 3:1
• Total flow rate 100mL/min
• continuous phase, Tris buffer, 10mM pH 7.4

The Pathfinder delivered highly repeatable, monodisperse LNPs that were 70nm in size with a narrow PDI of 0.13. The experiment was repeated in triplicate and samples were measured using the Malvern Zetasizer DLS system.

Encapsulation efficiency is about 97-98%. The AXF mini uses cross flow mixing which is a gentle mixing technique and prevents RNA degradation.

Process tunability and predictability

The two graphs above demonstrate the superior capability of the AXF mini to control the size of nanoparticles produced which can be critically important, especially for GMP manufacturing.

The plot on the right shows two formulations, plotted in red and green. Differences arise from e.g. concentration of DMG-PEG200 in the range 0.004 – 0.12 μmol which reduces size from 200 nm to 30 nm but both exhibit the same shape and therefore control. Although the PDI is still quite tight in both formulations but the formulation plotted in green is considerably larger (100-160nm) compared to the formulation in red (50-110nm). From this experiment you can quickly see it is possible to produce a 70nm particle by using a total flow rate around 100 ml/min. 

The plot on the left shows an experiment where the AXF mini was used to create two different formulations using two different flow rates. At 20ml/min the LNP size was 107 nm. When the total flow rate was increased to 200 ml/min, the LNP size reduced to 55 nm with a remarkably low PDI of 0.06.  

This data demonstrates the predicable scalability which is favourable especially for GMP manufacturing and regulators which means you can start to introduce process controls that are automated – process analytical technologies (PAT). Automated analysis of properties such as online particle size enables the option to automatically control any deviations in size and feed that back to control the pumps and optimise control and give the correct size again. This increases confidence in quality of production meaning throughput can be increased further. ATA Scientific offers several technologies to characterise LNPs.

Contact us for a demo today!

Live-cell imaging is important for many applications however limitations of conventional methods have constrained its routine use. Reliable live cell imaging requires an environment that keeps the cells functioning during the experiment while also being able to ensure the experimental method is not perturbing the cells and affecting the interpretation of the results. Here we discuss the growing popularity of automated live-cell imaging systems and highlight some key features to look for when selecting a live cell imaging system.

What is live-cell imaging?

Live-cell imaging is a microscopy-based technique used to examine living cells in real-time. It offers deeper insights into dynamic cellular processes such as migration, confluency and signaling and can reveal findings that might otherwise have been overlooked. Both brightfield and fluorescence-based live-cell imaging modalities support a range of different analysis needs.

How is live-cell imaging used?

Applications of live-cell imaging span basic research through to biopharmaceutical manufacturing. In a research setting, live-cell imaging can be used during cell culture to help define the best time for harvest, determine the senescence status of cells, assess drug treatments for cytotoxicity or detect and monitor phagocytosis. For example, Phagocytosis, is a process by which certain live cells, called phagocytes internalise foreign matter. This defensive reaction against infection is key in the study of immunology and plays an important role in immune responses, tissue homeostasis, and continuous clearance of apoptotic cells. Generally, phagocytotic activity is assayed using flow cytometry. However, this process only provides quantitative data and does not provide the means to monitor phagocytosis in real time. Performing fluorescence-based assays using the CELENA® X High Content Imaging System with pH-sensitive fluorescent particles, like pHrodo™ Green, can be an effective and efficient system for quantifying and monitoring apoptosis activity. For biopharmaceutical manufacturing, live-cell imaging has broad utility for process development and control throughout the production of biologic drugs and vaccines.

Limitations of conventional live-cell imaging methods?

Historically, live-cell imaging has involved manually monitoring cells by culturing them in a CO2 incubator. The culture vessel is removed several times to take images of cells over time using a digital microscope. This approach is labor-intensive and highly prone to human error, largely because it offers no means of finding the same position in the culture vessel. Fluctuating environmental conditions can also cause cellular stresses, which can compromise results. While benchtop imaging systems improve on this method, they are bulky and cumbersome, and often struggle to maintain a stable environment.

3 Factors to consider when choosing an automated live-cell imaging system

Automated live-cell imaging systems like the CELENA® X offer a flexible design that is smaller, faster and easier to use to meet both the demands of the drug discovery industry and the basic research needs of the smaller laboratory.

Multiple imaging modes that are affordable

Live cell imaging systems that offer both brightfield and fluorescence options for either time-lapse or real-time monitoring offer maximum flexibility. Celena X integrates an automated fluorescence microscope with quantitative image analysis software to process large datasets at an affordable cost. Its interface allows a user to run multi-well or multi-spectral experiments with capacity for multi-point imaging with only a few clicks. The microscope provides for a multitude of fluorescence cell imaging possibilities by supporting all objective magnification from 1.25x to 100x, both brightfield and phase contrast illumination, and with LED filter cubes.

Stable scanning performance and compatibility

The system is compatible with a wide range of cell culture vessels such as multi-well plates, dishes, flasks and slides to cover a wide variety of assay types. Depending upon the application, either image-based or laser-based autofocus methods can be used. The CELENA® X can be used for image confluency in McCoy cells seeded in 96-well plates over 48 hours with brightfield image-based autofocusing, demonstrating how the system can be modified and applied as a high throughput method for various cell-based assays. With laser-based autofocusing and multiple filter cubes, this method utilised the CELENA® X to image the dose-dependent effects of anti-cancer drugs throughout the cell cycle in HeLa cells seeded in 96-well plates, demonstrating how the system can be used in a multivariate drug screening process.

User friendly interface and 3D modeling

An analysis of high content images with large datasets can cause problems with most types of analytical software. Each new assay requires the creation of new modules, which can be challenging for lab staff and often involves IT staff. The CELENA® X provides an easy-to-use modular-design analysis software based on a powerful CellProfiler engine. Tens of thousands of images can be analysed automatically to obtain quantitative information with a complex software setting or optimisation.

Three-dimensional (3D) cell models can provide a more accurate representation of real cell environments than 2D cell culture systems but requires a different strategy of imaging and analysis compared to 2D cell culture. Organoids, for example, are organ-specific 3D cell models derived from human stem cells, designed to mimic the functionality and structure of human organs while 3D spheroids can represent a gradient of nutrients and oxygen between cells located in both outer and inner layers which is more relevant to physiological environments. These 3D models are notably useful for studying various types of cancers.

The challenge when imaging organoid and spheroid assays comes from organoids having multiple focal planes making it difficult to acquire in-focused images for multiple organoids. For live/dead cell viability of the single organoid, a different analysis strategy is required since individual cells in an organoid do not exist as a single live/dead status. To address this issue, CELENA X employs MergeFocus software module after acquiring Z-stack images from multi-channel fluorescence.

We invite you to use the Celena X automated live cell imaging system today and compare it for yourself.

Contact us to arrange a free trial.

ATA Scientific Pty Ltd
+61 2 9541 3500
enquiries@atascientific.com.au 
www.atascientific.com.au 

Product page link: – https://www.atascientific.com.au/products/celena-x-high-content-auto-cell-imaging-system/

Cell counting underpins numerous applications, spanning basic research through to the development and production of cell therapies. In recent years, manual cell counting methods have been replaced by the use of automated cell counters, which are both faster and more accurate, especially for complex sample types. Here we discuss 5 factors to consider when choosing an automated cell counter and share useful tips for instrument use to ensure accurate results.

1. Match the instrument specifications to the cell type

While cell-based research has traditionally relied on immortalised cell lines, it is increasingly common for more complex sample types to be used. These include primary cells, peripheral blood cells, stem cells, dissociated tumor cells, and even engineered T cells, which vary in terms of size, shape, and aggregation properties. To improve cell counting accuracy, many users have switched from manual, haemocytometer-based methods to counting cells with automated platforms. However, automated cell counters can only provide accurate results if they feature the right specifications for the cell type in question. Although an automated cell counter equipped with brightfield microscopy optics and a low-magnification objective represents a budget-friendly option for counting well-isolated, homogenous cell lines, an instrument capable of measuring fluorescence is often a better choice for counting clumpy samples or smaller cell types such as peripheral blood mononuclear cells (PBMCs).

2. Determine an appropriate sample dilution range

So, the question is, what factors should you consider when using an automated cell counter? A primary concern is the optimal dilution range, typically provided as cells/mL. This differs among instruments based on the size of the field of view (FOV), the magnification and the image sensor size. For counting accuracy, the sample should be diluted such that it falls within this range—if too dilute, counts will be inaccurate; if too concentrated, the instrument software will struggle to distinguish individual cells.

3. Consider using fluorescence staining for viability measurements

Most automated cell counters allow viability measurements to be performed using trypan blue staining and brightfield imaging. But, while this approach provides accurate results for homogenous samples like cancer cell lines, it is less reliable for more diverse sample types. For example, primary cells are often contaminated with large numbers of red blood cells, which will be mistakenly classified as dead cells after staining with trypan blue. Cellular debris and non-cellular particles can also be misidentified in this way, leading to data being artificially skewed. Fluorescence-based methods such as acridine orange/propidium iodide (AO/PI) staining provide greater accuracy than trypan blue, regardless of cell type, and are fast becoming a preferred method for measuring cell viability.

4. Maximise the counting volume

Once samples have been prepared for counting, they are loaded onto a chamber slide; this functions to provide a fixed volume measurement based on the chamber height. Because the chamber height is set by the slide manufacturer, scanning multiple FOVs is the best way of increasing cell counting volume to achieve greater accuracy and precision. Modern instruments equipped with an automated scanning stage can count volumes of up to 5.1 µL, which is 10-fold higher than a conventional haemocytometer measurement.

5. Optimise the counting protocol

The most common error when counting cells is incorrect focusing, which, for brightfield microscopy, can lead to poor discrimination of live cells from dead cells. If an automated cell counter offers only manual focusing, it is recommended that users obtain a bright spot at the center of each live cell for brightfield imaging to produce an accurate count. Fortunately, most modern automated cell counters feature an autofocusing function, which can minimise focus-related issues when running in brightfield mode. Alternatively, since fluorescence imaging is relatively insensitive to focus, using a fluorescence cell counter with an autofocus function will eliminate this problem.

Another widespread mistake is failure to optimise the cell counting algorithm, also known as the counting protocol. This is proprietary to each automated cell counter and is used for image preprocessing, object finding, and object classification. As it would be impossible for a single counting protocol to cover every cell type, most algorithms allow the end user to perform further optimisation. Factors to consider here include the cell size, spot brightness, cell detection sensitivity, roundness, noise reduction, and fluorescence intensity, all of which should be carefully tailored to the diversity of cell types within the sample.

Introducing the LUNA™ Automated Cell Counter

The LUNA™ automated cell counter family from Logos Biosystems provides fast, accurate counting for a broad range of cell types, including PBMC and CAR T cells. Dilution of highly concentrated samples and replicate counts at low cell concentrations are no longer necessary when using the LUNA series of Automated cell counters.

The LUNA-FX7 is the newest member of the LUNA Automated Cell Counter family that provides unmatched cell counting accuracy, dual fluorescent and brightfield detection, advanced declustering algorithm, precision autofocus and 21 CFR PART 11 compliance. It has built-in quality control features and precise validation slides for monitoring QC and bioprocesses. LUNA-FX7 meets a broad range of counting applications from confirming cell quantities for single-cell sequencing to highly accurate cell therapy dose.

Key benefits:

• Larger counting volumes, up to 5.1 µL for high accuracy (1% CV)
• High throughput using either 8- or 3- channel slides.
• Bioprocess package that can monitor individual and specific batches.
• Innovative CountWire™ package for 21 CFR Part 11 compliance.

Higher throughput. Offering a variety of slide options, the LUNA-FX7™ utilises a counting volume of up to 5.1µL, lowering error and CV for each count.

We invite you to use the LUNA-FX7 Automated Cell Counter today and compare it for yourself.

Contact us to arrange a free trial.

ATA Scientific Pty Ltd
+61 2 9541 3500
enquiries@atascientific.com.au 
www.atascientific.com.au 

Product page link:
https://www.atascientific.com.au/products/luna-fx7-automated-cell-counter/

Australia’s Additive manufacturing (AM) industry is off and running, transforming the way we produce and distribute goods. Parts which required multiple components to assemble manually can now be produced more viably using AM in a one-step build process. It removes the need for complex shipping arrangements to move instruments from place to place, relying instead on digital files to print products on-demand. In the heat of the COVID-19 pandemic, 3D printing stepped up to become a vital technology to provide solutions to severe disruptions in supply chains ranging from personal protective equipment (PPE) to emergency dwellings to isolate patients. From aerospace to automotive engineering, and from the medical to the dental industry, AM is an evolving technology revolutionising industries across the country.1

Backed by the Australian Government ‘Modern Manufacturing Strategy’ as well as the funding available for the sector, has enabled new manufacturing-focused research facilities that work alongside industry.2 With new opportunities to deliver cutting-edge R&D in AM and materials processing, highly complex or previously unachievable products can be created quickly and efficiently for the global market. CSIRO established Lab22 with a vision to grow a new manufacturing industry as Australia’s Centre for Additive Innovation citing a recent focus on critical mineral and hybrid manufacturing.3 The University of Sydney and GE Additive have also joined forces to collaborate on R&D topics and demonstrate AM technology via the new Sydney Manufacturing Hub.4 These and many other AM research facilities in Australia underpin a growing AM industry helping to build sovereign capabilities. Instead of sending processed ores overseas and importing them back as powders, Australia can forge on finding ways to turn minerals into new AM innovations. 

But as exciting as the possibilities are in AM, the process itself is not without its challenges. Problems with final product consistency and a narrow range of expensive raw materials are some of the biggest obstacles to the widespread adoption of AM. In processes that use powder as the raw material, for example, just one particle could contaminate the rest of the material, impacting the overall quality of the end product. To maintain consistent high quality in these components, producers need to ensure that their input materials are carefully monitored and optimised. 

Why particle characterisation is critical 

Key to developing and manufacturing high quality materials with the required functionality and performance is understanding the relationship between material structure and material properties. From metals and polymers to composites and ceramics, monitoring the particle size and shape is important to ensure the powder supply is consistent and meets specifications. Beyond quality control, it also plays a vital role when investigating novel alloys or composites or developing a new AM process.

Below are two key analytical tools that support additive manufacturers with material characterisation.

The use of laser diffraction for particle size distribution

Particle size distribution is critical for powder bed AM processes since it affects powder bed packing and flowability which in-turn impacts on build quality and final component properties. The Malvern Mastersizer 3000 uses laser diffraction, an established technique for measuring the particle size distribution of metal, ceramic and polymer powders for additive manufacturing, and is employed by powder producers, component manufacturers and machine manufacturers worldwide to qualify and optimise powder properties. A complete high-resolution particle size distribution is provided in a matter of minutes (from 10 nm to 3.5 mm) using either wet or dry dispersion. The technique can also be integrated into a process line to provide real-time particle sizing.5

Automated Image Analysis for particle shape and composition

Powder bed density and powder flowability are influenced by particle size and shape. Particle morphology is therefore, another important metric for powder bed additive manufacturing, with smooth, regular-shaped particles preferable as they can flow and pack more easily than those with a rough surface and irregular shape. The Malvern Morphologi 4-ID provides automated optical image analysis to classify and quantify the size and shape of metal, ceramic and polymer powders. The fully integrated Raman spectrometer also enables component-specific morphological descriptions of chemical species.

The Phenom ParticleX AM is a specialised high-resolution desktop scanning electron microscope (SEM) dedicated to optimising AM metal powders and final product quality. By combining an imaging resolution of <8nm and magnifications up to 200,000x together with X-ray analysis (EDS) for elemental composition, properties such as structural integrity, print resolution, surface uniformity, phases and the presence of impurities or defects can be determined to contribute unique insights not possible with other systems. A scanning area of 100x100mm, grants a large degree of freedom to image and assess the size and shape of whole parts or sections of a larger component simultaneously. This fully integrated system is simple to operate and eliminates the need for outsourcing for quality checks, speeding up time-to-market. 

We can go further together

At ATA Scientific, we don’t just sell our instruments – through collaboration with a broad range of industries and academic institutions, we play a key role in the AM ecosystem. We support our customers by providing optimal material characterisation techniques used in AM together with key insights into the application, measurements and analysis to fully understand material behaviour. 

Contact us for more information today!

ATA Scientific Pty Ltd
+61 2 9541 3500
enquiries@atascientific.com.au
www.atascientific.com.au

References

1. Additive manufacturing: could it drive global success for Australian businesses?
   https://www.materials-talks.com/how-additive-manufacturing-can-take-your-industry-to-new-heights/
2. Additive manufacturing and critical minerals come together at CSIRO’s Lab22 – CSIRO
   https://www.exportfinance.gov.au/resources/article/additive-manufacturing-could-it-drive-global-success-for-australian-businesses
3. Additive manufacturing and critical minerals come together at CSIRO’s Lab22 – CSIRO
   https://www.csiro.au/en/work-with-us/industries/mining-resources/Resourceful-magazine/Issue-27/Lab22-and-critical-minerals
4. $25M Sydney manufacturing hub launches to drive state wide innovation
   https://www.3dprintingmedia.network/25m-sydney-manufacturing-hub-launches-to-drive-statewide-innovation/
5. Characterising material properties for powder additive manufacturing
   https://www.materials-talks.com/characterizing-material-properties-for-powder-additive-manufacturing/

As RNA science continues to make headlines around the world as the new way of making safer, more targeted medicines, it has sparked a wave a new studies that offer significant potential not only as broad-spectrum vaccines but also treatments for cancer, genetic and autoimmune diseases. This is quite extraordinary, given prior to the COVID-19 outbreak, few people outside the RNA research community were aware that this technology even existed. That Australia is still unable to manufacture mRNA vaccines – despite their proven success against the SARS-CoV-2 virus – presents a huge opportunity.

The COVID-19 pandemic brought together world-leading researchers in a mass response from across many fields, diverting resources to deliver a vaccine that could help save millions of lives. Scientists stepped up and worked tirelessly despite the myriad of challenges, the biggest one of which was the challenge of vaccine supply. Recognising this challenge and the need for building local capability, led to a humble yet exceptionally talented scientist from Iceland to step up and run the first RNA institute in Australia.

Australia’s RNA capability strengthens as UNSW RNA Institute opens

Professor Pall (Palli) Thordarson, an award-winning researcher and chemistry professor at UNSW Science, is set to kickstart RNA capabilities here in NSW and finally launch our Genetic Medicine Ecosystem. Fuelled by his interest in the interface between chemistry and biology, Palli has always been fascinated by the role of RNA and now, this molecule is the focal point of his work. RNA science seems like an overnight success, but it has been decades in the making and holds huge potential for making critical contributions toward advancing human health. The facility will allow scientists to connect and network with industry partners and other collaborators to meet research and manufacturing needs.

The importance of driving onshore advances in RNA research and therapies

Today, the NSW Government together with 14 universities that constitute the NSW RNA Bioscience Alliance and the dozen research organisations within the NSW RNA Production Research Network are now involved2. The goal to create a national Genetic Medicine manufacturing facility in Australia, was captured during an early meeting of the Australian RNA Production Consortium (ARPC). It is inspiring to see so much occurring around the nation knowing each of the original members of the ARPC are tirelessly working to build this, in a fashion that is not simply replicating facilities in each state, but to build a collaborative web of skills and infrastructure. These people have changed the future for scientific research in Australia – thank goodness for this cognisant few!

It seemed puzzling to find a chemistry expert amongst so many RNA giants however, as time passed, it had become clear the sheer value someone like Palli had in such a forum. Palli’s fascination with RNA began to emerge early in his PhD, “I remember looking at RNA and RNA-based systems biology as a PhD student and thinking, ‘These are the most fascinating chemical machines in the world’.” Palli has long tried to connect the dots between the pure chemistry world and the biological sciences and just how that chemical machine results in a biological translation.

As we spend inordinate efforts to educate and develop a constant crop of scientists, from high school to early career researchers, we hear the same things from PhD students today as we did back in the 1980’s and I suspect before- “we can’t find work as we are either too inexperienced or we are over-qualified!” Such enormous investment into this ecosystem can only help resolve much of this. 

Whilst there is a great deal of investment in Victoria and some in QLD, there are synergies between the research institutes and many collaborations. The global nature of science has throughout history proven there are no boundaries, no borders and only one tribe – the science tribe – we see beyond nationality – and we travel the planet in a quest for knowledge, going to where the research is. To be part of this global community Australia needs vision such as shown by Palli. Without this, we could never attract the best of the best to come to Australia, nor would we retain our global giants.

Next generation technologies will nurture a wide range of genetic medicines

Being privy to much of the research and seeing technologies you supply have material impact in the future lives of potentially millions of people is inspirational. Consider this technological revolution answers the question we didn’t even know existed – to solve a problem we didn’t know we had. As Palli often states- the missing link to all this manufacturing ecosystem is the humble Lipid Nanoparticle. Now humble should not elicit the thought it is simple, far from it. However, it has been made a great deal simpler by the NanoAssemblr platform3. These technologies are unique. They create very small particles, quickly and repeatably. They encapsulate a payload, such as mRNA, a peptide, protein, or small molecule for the delivery to the cell by stealth. Imagine you are a cancer researcher and you have figured out the mechanism of how the cancer cells are replicating and where they stem from. Using a Nobel prize winning technique called CRISPR you edit the RNA – how do you do this? Hitch a ride inside a Lipid Nanoparticle, slip into the tumour and stop it in its tracks! This is elite science happening right here in Sydney with team collaborations across the nation. This is the key – we can do wonderous science that will save lives – this is what drives the scientist. The government sees the billions in revenue and the possibility of jobs, jobs, and jobs. It makes ethical and economic sense! 

The benefits of RNA research are far-reaching

For those who assume it is just medical, think again. The Agricultural industry is set to benefit from multi millions in investment with the help of Palli to aid in biosecurity, disease prevention and control – think lumpy skin or foot and mouth disease in cattle. It is hard to see any downside to what Palli has orchestrated. It is visionary and all power to UNSW backing him to run down this path. Nice to see Palli back on the farm.

ATA Scientific collaborates with thousands of scientists throughout Australia and New Zealand providing solutions for scientific challenges. We are constantly adding novel technologies that help pose the obscure questions needed to advance science. Collaborate with us today www.atascientific.com.au

References

1) https://newsroom.unsw.edu.au/news/science-tech/australias-rna-capability-strengthens-unsw-rna-institute-opens Website Accessed 19 July 2022
2) https://newsroom.unsw.edu.au/news/general/unsw-celebrates-119m-funding-support-rna-research-across-14-universities Website accessed 19 July 2022
3) https://www.precisionnanosystems.com/ Website accessed 19 July 2022

The need to build new energy storage solutions to address the increasing global demand has helped drive a power revolution in battery research and technology. Lithium-ion (Li-ion) batteries are predicted to play a key role in the trend toward renewable and sustainable industrial electrification solutions. As fossil fuels are phased out and CO2 regulations become more stringent, the increase in demand to provide ever more lightweight, low-cost, safe, high-power and fast-charging batteries has accelerated advances in battery technology.

Access to the right tools and technologies can help optimise R&D and production cycles, investigate causes of battery failure, improve safety, and speed up time-to-market, to keep technological progress moving in sync with modern global demands. Here we discuss a complementary set of physical, chemical, and structural analysis solutions designed to enable rapid, high-precision analysis of particle size and shape distribution plus elemental composition of battery materials for the entire process from research through to production.

With the Mastersizer 3000, particle size can be rapidly analyzed with ease, while the Morphologi 4 can image and classify thousands of particles automatically with high statistical accuracy. Our Zetasizer can analyze the zeta potential of a dispersion and also the size and agglomerate state of nanosized materials. Phenom XL G2 scanning electron microscope (SEM) is an unrivalled technique that allows users to observe the 3D structure of powders and electrodes and also identify elements and the presence of contaminants.

The importance of particle size measurement

The performance of a battery can be characterised according to the amount of energy that it can store or the amount of power that it can produce. The maximum battery power can be increased by decreasing the particle size of the electrode material and increasing the surface area. Battery power is determined by the rate of reaction between the electrodes and the electrolyte, while storage capacity is a function of the volume of electrolyte within the cell. These properties are intrinsically linked to the intercalation structure and particle size of the electrode particles, which determine how well the mobile ions are taken up and released by the electrode. Particle size distribution and particle shape influence particle packing, hence the volume of electrolyte that can be accommodated within the interstitial voids of the electrode, which affects storage capacity. As a result, a mixture of coarse and fine particles is often used in the electrodes to increase surface area, whilst also controlling the overall packing fraction of the electrode material to allow good contact between the electrode and the electrolyte.

Particle sizing of electrode materials is commonly performed using the Mastersizer 3000 which uses automated laser diffraction technology. With a measurement range that runs from 0.01 to 3500 µm, the Mastersizer is the particle sizing technology of choice for most battery manufacturing applications – starting from precursor to the final milled electrode materials.

Figure 1 Particle size distribution of three batches of NCM cathode materials synthesized with different processing parameters

Figure 2 Particle size distribution of three batches of synthetic graphite synthesized with different heating conditions

The Malvern Insitec online process systems deliver real-time monitoring of particle size for automated process control. These can be used for either the monitoring of particle size evolution in precursor slurry or in the control of electrode material size right after the mill. Smaller particles in electrode slurry production can be prone to agglomeration and/or flocculation, resulting in uneven electrode coatings and ultimately compromising the electrochemical performance. Aggregation and stability can be monitored by measuring zeta potential (particle charge) using the Malvern Zetasizer Ultra. A low zeta potential will indicate particles likely to aggregate whereas a high zeta potential will form a stable dispersion. The Malvern Zetasizer Ultra builds on the legacy of the industry-leading Zetasizer Nano Series adding high-resolution sizing (Multi-Angle Dynamic Light Scattering) and particle concentration capabilities.

The importance of measuring porosity

Porosity is an important parameter both for the separator and for the electrolyte to transport lithium-ions between the anode and cathode. By controlling porosity, higher intra-electrode conductivity can be achieved to ensure adequate electron exchange as well as sufficient void space for electrolyte access/transport of lithium-ions for intercalation of the cathode. Higher porosity means less heat generated in the cell and greater energy density. However, excessive porosity hinders the ability of the pores to close, which is vital to allow the separator to shut down an overheating battery. Therefore understanding the porosity of the electrode materials is important to guarantee the right ion accessibility and charging speed.

Recognised as the most advanced instrument in the field for material surface characterisation the Micromeritics 3Flex has become a crucial tool for the battery industry. The 3Flex is a high-performance adsorption analyser designed for measuring surface area, pore size, and pore volume of powders and particulate materials. Analysis of BET surface area, pore volume and pore size distribution helps to optimise battery components.

The Micromeritics AutoPore V Series utilises Mercury Porosimetry, a technique based on the intrusion of mercury into a porous structure under controlled pressures, to calculate pore size distributions, total pore volume, total pore surface area, median pore diameter and sample densities.

The importance of measuring surface area

Increasing the surface area of the electrode improves the efficiency of the electrochemical reaction and facilitates the ion exchange between electrode and electrolyte. Lower surface area materials are better suited for improved cycling performance of the cell resulting in longer battery life. High surface area presents some limitations due to the degradation interaction of the electrolyte at the surface and resultant capacity loss along with thermal stability. Nanoparticles hold promise to increase surface area without capacity loss by permitting shorter diffusion paths for lithium-ions between the graphite particles which facilitates fast charge and more efficient discharge rates and improves the capacity of the battery.

The Micromeritics TriStar II Plus is an automated, three-station, surface area and porosity analyser. MicroActive software allows the user to overlay a mercury porosimetry pore size distribution with a pore size distribution calculated from gas adsorption isotherms to rapidly view micropore, mesopore, and macropore distributions in one easy-to-use application.

The importance of measuring particle shape

Shape will affect the electrode coating in terms of packing density, porosity and uniformity. Spherical shaped particles will pack more densely than fibrous or flake shaped particles. The average strain energy density stored in a particle increases with the increasing sphericality. Fibrous and flake shaped particles are expected to have a lower tendency for mechanical degradation than spherical-shaped particles. Automated imaging using the Malvern Morphologi 4 is commonly employed for particle shape analysis of electrode materials but can also be coupled with Raman spectroscopy to give particle-specific structural and chemical information.

The importance of analysing chemical composition 

Deviations in chemical composition or impurities in electrode materials can significantly affect final battery performance. For this reason, chemical composition and elemental impurity analysis are an integral part of the battery manufacturing process. Simple to operate and fast to learn, the Phenom XL G2 scanning electron microscope (SEM) is an unrivalled technique that allows users to observe the 3D structure of electrodes after production; the size and granulometry of raw powders; the size of pores and fibres in insulating membranes and the response of materials to electrical or thermal solicitations. Using fully integrated X-Ray analysis (Energy Dispersive Spectrometer, EDS) the distribution and identity of elements including the presence of contaminants in the battery sublayer can quickly be revealed.

The Phenom XL G2 is the only SEM that can be placed within an argon-filled glovebox, allowing users to perform research on air sensitive lithium battery samples.

The future of batteries

Driven by our need to reduce greenhouse gases with renewable energy and portable communication devices, the Li-ion battery market is growing at 14% Compound Annual Growth Rate (CAGR). Deutsche Bank forecasts lithium-ion batteries will account for 97 percent of battery use in energy storage alone by 2025. Most automotive companies are now investing in batteries and are in a race to patent critical next-generation battery technologies and battery management systems. Companies related to fossil energy or mining are also entering the battery value chain.

Australia is well positioned to capitalise on the significant opportunities presented, having the world’s third-largest reserves of lithium and is the largest producer of spodumene (mineral source of Lithium). Australia currently produces nine of the 10 mineral elements required to produce most lithium-ion battery anodes and cathodes and has commercial reserves of graphite – the remaining element. Australia has secure access not only to all the chemicals required for lithium-ion battery production including precursor, anode, cathode, electrolyte materials but also the knowhow through various research groups at our universities and institutions such as the CSIRO, meaning more advanced batteries can be manufactured locally. However, as demand for lithium batteries continues to increase, it will eventually outstrip supply, so we need to think beyond Lithium to other technologies that can deliver safer, more abundant and cheaper materials (such as sodium, zinc or vanadium) to store renewable energy.

At the University of Wollongong , the Smart Sodium Storage Solution (S4) Project aims to develop sodium-ion batteries for renewable energy storage. This ARENA-funded project builds upon previous research undertaken at the University of Wollongong and involves three key battery manufacturing companies in China. Gelion, a spin-off company from the University of Sydney, is developing gel-based zinc-bromine batteries. The technology uses a unique gel electrode that transforms zinc-bromide technology into a high-efficiency non-flow battery.

Energise your battery research with ATA Scientific

Whether you are a battery component manufacturer looking for greater process efficiency and better quality control, or a researcher striving to determine the performance parameters of newly emerging battery materials, our solutions will offer you the new levels of insight and control needed to power the production of superior quality batteries. Contact us via phone (+61 2 9541 3500), or through our website for a demonstration or quote today!