Fast Prototyping of Fluidic Devices Using Fluidic Factory 3D Printer 28 Jun 2016
Quick fabrication of microfluidic devices has always been challenging: existing fabrication methods for microfluidic devices such as injection moulding, micromilling and bonding involve long and costly fabrication processes. 3D printers using SLA and SLS methods on the other hand are not able to print embedded microchannels in a single step, hence leading to relatively long fabrication times, high costs as well as limitations to material selection. Other FDM printers have a critical limitation in the milli- and micro- fluidic industry: structures created with such devices inevitably leak when trying to contain fluid.
To address these challenges, Dolomite Microfluidics recently introduced Fluidic Factory 3D printer, capable of fabrication of fluidically sealed devices in just minutes. Furthermore, the printer uses COC – and FDA approved, transparent, robust polymer suitable for biological applications.
We spoke to Mark Gilligan, CEO of Dolomite Microfluidics to discuss the benefits of the printer.
JR: Why is the Fluidic Factory 3d printer so unique?
MG: Fluidic Factory is the only platform in the 3D printing industry able to 3D print with COC. COC is FDA approved material for implantables, and is often the material of choice of diagnostic device developers. It is also suitable for biological applications and features great transparency as well as very low water absorption.
The 3D printer has been designed specifically for fluidic sealing in one step.
The intelligent software analyses the 3D geometry of the device to be printed and identifies the internal voids and surfaces. It then creates print paths from the inside of the device outwards and the print head deposits filaments in a continuous, leak-proof manner.
The FDM (fluid deposition modelling) method melts the polymer at high temperatures and ejects it through the nozzle to the print bed, which then solidifies at temperatures below 75°C.
The printer nozzle head is extremely accurate in its movement and allows precise features to be printed (with an accuracy of less than 1 µm). The nozzle is also disposable, which ensures that printing quality is preserved.
JR: What benefits does Fluidic Factory have when compared to traditional FDM printers?
MG: Many traditional FDM printers deposit the molten polymer beads in circular cross sections, which results in leak paths and, consequently, an inadequately sealed device. Fluidic Factory is unique in dispensing a ‘squashed’ section bead which eliminates these leak paths, creating a reliably sealed device that will contain any fluids passed through it.
JR The printer has a very small footprint, but how easy it is to set up and use?
MG: Fluidic Factory is a true out of the box solution – get started in just a few minutes! As the footprint is very small, it can be used either in an office or a lab and can be easily transported.
We have designed the printer so it really is easy to use with a very simple workflow. Fluidic Factory enables the user to have ultimate design creativity by letting them design a device using virtually any CAD software on their PC. Design files are saved as .stl files, which carry information about the surfaces of the model which then will be uploaded to the Fluidic Factory Software. The Software creates a graphic illustration of the device and calculated optimum print paths to ensure fluidic sealing. Once the file is ready – it can be transferred to Fluidic Factory via a USB drive.
The design can be then selected for print using the touch screen. Once printing has started, the screen displays the time left to print, real-time status and filament use, with an auto-alert when the COC is running low. The polymer reel contains 60 m of polymer and can be changed in seconds.
When printing has finished, Fluidic Factory will indicate this, which allows the user to remove the print tray and immediately insert another one to begin printing a new device whilst waiting for the completed one to cool down off the machine.
JR: What sort of devices can I create using Fluidic Factory 3D printer?
MG: A very wide range! Unique devices such as 3 dimensional mixers, non-rectangular chips, unique channel geometries and features not possible using etching, embossing, moulding or machining can be created using Fluidic Factory. Circular, triangular or rectangular channel geometries can be printed (dependant on the mode of printing) and other cross-sections are also possible. By following some simple rules to optimize your design, the possibilities to create unique fluidically sealed devices quickly and in a cost effective way are endless! JR: Will Fluidic Factory evolve to enable future functionality?
MG: Yes! Fluidic Factory features a replaceable head and bed and upgradeable software to enable future functionality e.g. on-line chip design files shop, printing alternative polymers high definition printing, micromilling, fluid dispensing and bio-printing.
JR: What else should I know about Fluidic Factory?
MG: We believe Fluidic Factory 3D printer is the future of fast prototyping for micro-and milli fluidic industry and with additional future functionality, a cost-effective, efficient and reliable alternative to traditional microfluidic prototyping alternatives.
Examining the Relationship of Chemicals and the Circular Economy 22 Jun 2016
As knowledge of the impact of chemicals on the environment and human health increases, regulations to control hazardous chemicals increase. The Helsinki Chemicals Forum (HCF) promotes chemicals safety and management globally, and is closely aligned with the European Chemicals Agency (ECHA) REACH (Registration, Evaluation, Authorisation and Restriction of Chemicals) regulation. This year HCF 2016 provided five panels that focused on specific issues concerning REACH: The circular economy, perfluorinated chemicals, global data sharing, plant safety, and SVOC’s in the construction industry.
The overall theme throughout HCF 2016 was on economic development and sustainability with a particular emphasis on the opportunities and challenges the circular economy poses for chemicals regulation. A circular economy is an industrial economy that is producing no waste or pollution. It is a move away from the “take, make, dispose” manufacturing model, and replaces cradle-to-grave design with cradle-to-cradle, which is the driving philosophy behind the field of sustainability.
Thus while ECHA strives to regulate hazardous substances, it is also promoting the use of alternative non-hazardous options that eliminate the need to register at all, resulting in reduced hazardous chemical use, reduced waste and reduced risk: a win-win situation for everyone. 11th Stakeholder’s Day Preceded HCF Conference
Preceding the conference was the 11th Stakeholder’s Day, which attracted 300 attendees from more than 30 different countries, including those in the European Union as well as India, Russia, Japan, Turkey and the USA. The day was devoted to discussing how to achieve a successful REACH registration, how to ensure high dossier quality, and how registrations are used.
Geert Dancet, ECHA’s Executive Director, opened the venue, and announced ECHA’s recently published 5-year report that examines how chemical use has changed as a result of REACH. According to the report, 168 substances of very high concern (SVOC) have been identified, 31 of which require review and 20 of which are restricted. The report concludes that as a result of REACH there are more SVOC controls and that chemical use now delivers safer conditions for manufacturers and downstream users.
The next REACH deadline, May 31, 2018, by which all chemicals must be registered, was emphasized. This particular deadline is expected to swamp ECHA with up to 60,000 dossiers, about five times more than they received in 2013. Up to 25,000 substances are expected to be registered as compared to just 3,000 in 2013. This increase in volume is anticipated to come from small to medium sized entities (SMEs), including more registrants without co-registrants. Guidelines were published at the end of May 2016 to provide additional details about data sharing and low-risk, low-tonnage substances.
ECHA also introduced a new generation of IT tools for IUCLID, Chesar, and REACH IT designed to reduce the number of user manuals and drive ease of use, with a focus on streamlining the registration process. All three tools are better integrated and less confusing to navigate that previous iterations.
As REACH registration matures, ECHA is paying attention not just to substance identification but also use descriptions and hazard assessment conclusions.
All the presentations focused on enabling organizations to better address the REACH regulation. One presentation in particular stood out. Janet Greenwood, secretary for the UK’s Chemical Regulations Self Help Group, provided a case study concerning the greatest registration difficulties, including finding out who the lead registrant is for an existing registration and what to do if no lead registrant exists. She also examined the issue of registration costs. For instance, an SME with a turnover of £3.6M, net profit of £110K and 21 staff with 8 substances to register as the lead and only registrant will incur estimated costs of £60K per substance for a total of £400K. If the SME is a member registrant, the estimated costs drop to £33K per substance for a total of £264K. This is obviously an unsustainable number either way. The options are to register only what the SME can afford, but lowering the Letter of Access costs would reduce the impact of REACH significantly. Greenwood said “the costs of REACH have to be weighed against access to the market and thus doing business in the EU. To offset these costs there will need to be an increase in revenue in the EU under REACH.” Another option would be to increase the cost of the product, but this may not always be feasible. Greenwood’s presentation is available along with all other Stakeholder Day presentations at http://echa.europa.eu/news-and-events/events/event-details/-/journal_content/56_INSTANCE_DR2i/title/11th-stakeholders-day.
Triskelion’s Hans Marquet pointed out that knowing your substance involves more than knowing the melting point, viscosity, relative dew point, etc. In fact, while many substances perform well at their specified task, but there is often little data about hazard attributes. This lack of data is one of the greatest impediments to registration, and not just the responsibility of the manufacturer, but also required from the organization that uses the substance in their own products. Much of this data should be on the Safety Data Sheet (SDS) but is often not, resulting in the greatest source of REACH non-compliances.
These challenges contribute to the difficulties faced by ECHA in understanding hazardous chemical use in the EU, by substance manufacturers in understanding the substance, and by research organizations that use the substances in their development processes in understanding the impact of the substance on their chemicals-based end-products.
HCF 2016 Examined Chemicals Impact
During the two-day conference that followed Stakeholder’s Day, keynote speaker Achim Halpaap, Head of UNEP’s Chemicals and Waste branch, provided some telling statistics about chemicals management and the importance of the move toward a circular economy. He stated that the world economy and global market is expected to quadruple between 2010 and 2050. Halpaap emphasized that there are fundamental structural changes occurring in the chemical industry what with China becoming a dominant player and large multinationals becoming larger. He also pointed out that there is a double standard: chemicals that are classified as unsafe in one part of the world should not be “safe” in another, citing lead in paint as just one example. UNEP’s booklet “Global Chemicals Outlook II 2018” examines this subject in detail.
The European Commission’s Bjorn Hansen led the panel on the Circular Economy, drawing attention to chemicals substitution, reduction, non-toxic material cycles, risk management, recycling and cross-border circulation of raw materials. All of the panelists emphasized the challenges of changes to business models as a result of increased regulatory oversight. One panelist, Michael Warhurst, Executive Director of CHEM Trust, pointed out the issue of Bisphenol A (BPA) in thermal paper from cash register till receipts being recycled into pizza boxes and thus creating a inadvertent downstream issue of food contamination by an endocrine disrupting chemical. Hansen stated that “as the world increases from 1B to 2B consumers, there will be an increase in use of consumer goods. As knowledge develops, we find more and more hazards in those goods. Thus the transition that needs to happen needs to stimulate innovation while eliminating chemicals coming in to the waste stream.” There are opportunities for research organizations within these parameters, specifically the reduction in hazardous material waste can lead to significant cost savings for the organization. Hansen added that “REACH is the start of this process and has sensitized industry to think about safety and driven awareness by a broader community that everything is chemicals-based.”
Each of the subsequent panels addressed specific concerns associated with these challenges. From the fact that less than two percent of perfluorinated chemicals are registered under REACH to more efficient sharing of regulatory data globally to case studies on how to improve plant safety during research and production processes, HCF 2016 examined challenges and solutions to chemicals use and management from a number of perspectives.
Next year HCF returns to Helsinki’s Messukeskus Convention Centre from 8-9 June 2017. For details, visit http://www.helsinkicf.eu or http://echa.europa.eu/regulations/reach.
Following the discovery of nanoscale submarine, scientists have made further development on it to make it better fit the research condition in labs. Being florescent traceable is one of the prominent advances in the new round of optimization.
The nano vehicle was developed by chemist from Rice University in 2015. Each nanosubmarine is a single-atom molecule, powered by a motor that makes use of the ultraviolet light as energy source, and the motor is equipped with a propeller, functioning together, the Nanosubmarine can move at a relatively fast speed in solutions. As the motor can spin at a pace of one million per second, Nanosubmarine can move at about 18 nanometers per second.
By taking advantages of ultraviolet light, the nano molecule won’t produce any toxic chemicals as previously developed nano machines did, which widens its usage in both chemistry and biology researches, even in practical applications.
With one limitation that the activities of nano-vehicle couldn’t be observed well, the new tech was not employed so commonly in labs. The new development was aimed to address the issue, and provide image information for its function route in solutions. Then Fluoresce was taken into consideration as it’s widely taken advantage of in protein studies in the form of fluorescent labeling of proteins
Labeled with fluoresce, the nanosubmarine now can be watched under one-molecule microscopes for one and a half seconds, which can be extended into a 30 frames length video, enough for a thoroughly observation.
The most potential use for the nano scale vehicle should be drug deliver and the research team will go on with their studies to probe about its ability in communicating with cells. However, the Nanosubmarine still can’t be steered at present, but with the enhanced diffusion, chemists are able to move them out in the solutions consist of similar molecules in size.
With fluorescent labeling, the nanotech can be directly tracked in the form of images, which is just one case of the application of the labeling tech. For professionals in the field, they won’t be unfamiliar with the method as they are frequently applied in protein tracking. The tech will be more mature with the related tech is still getting optimized.
Author Bio BOC Sciences is a professional chemistry services provider. Fluorescent labeling of proteins is newly launched at the company with its increasing needs by researches. In total, five types of fluorescent labeling are available, including Enzymatic labeling, Chemical labeling, Genetic labeling, Protein labeling, Quantum dot labeling.
Phasefocus have recently launched Livecyte™ - a unique system for live cell analysis – or kinetic cytometry - which will enable a deeper understanding of the biology in many application areas including immunological, neurobiological, cancer and basic cell biology research.
We spoke to Tracey Zimmermann, VP of Global Sales at Phasefocus to learn more about the application areas for Livecyte™ and what unique new insights the technology will enable researchers and drug developers to reveal about the behaviour of cells’
JR: Could you tell us about the key features of the Livecyte system?
TZ: Livecyte is a unique imaging system for live cell analysis which uses an optimized version of Quantitative Phase Imaging called Ptychography (tie-cog-rafee), to enable Kinetic Cytometry: the automatic tracking and analysis of phenotypic and kinetic behaviour of individual cells and cell populations over hours or days. The system produces high contrast, high fidelity images which are artefact free, quantitative and can be tracked with ease across the whole time-course, without the need for cell labelling or high intensity light imaging. Since the power of the illumination laser is thousands of times less than that used for traditional fluorescence light microscopy, cells suffer far less phototoxic shock-a particular advantage for fragile cells such as stem cells or primary cells.
JR: What challenges will this new system help researchers to overcome?
TZ: Time-lapse imaging is a fundamental tool for studying cellular behaviours. Live cell studies often involve fluorescent cell labelling techniques which produce high contrast images, but are compromised since fluorescent dyes require high intensity illumination to visualise their location and this can disturb normal cell functions, particularly over extended time-courses. The label free approach which Livecyte affords, allows longer observation of “happy” cells which have been minimally perturbed.
Another challenge facing time-course assays can be focal drift which can affect image quality, which in turn makes it very difficult to track cells. Livecyte’s ptychographic imaging technique ensures that cells are always in focus and since the system delivers a continuous field-of-view which is arbitrarily large (and can be up to a few millimetres in a single image), the need to stitch images is removed. Additionally, imaging continuously over multiple regions (at different magnifications if desired), within a single well of a multi-well plate or multiple regions within each well of a multi-well plate, ensures maximum productivity from each experiment.
Cells in culture and particularly in co-culture, are heterogeneous in nature and averaging these populations in assays may mask interesting characteristics of those cells which do not represent the majority, but may be significant in determining the ultimate behaviour of a population. Since Livecyte allows single cell analysis within cell populations, data on the typical, the median and the average cell can be obtained, whilst also allowing identification of outliers within one time-course experiment. This is exciting since in drug discovery, the outliers-clinically and drug resistance wise-could be the most interesting.
JR: Phasefocus recently had a paper published in Nature Scientific Reports validating this new technology. Could you tell us about the significance of the results the paper described?
TZ: The paper was published in conjunction with researchers from University of York and highlights the power of ptychography for long-term imaging of complex primary neuronal cultures in conditions which are effectively “closer to real life”: http://www.nature.com/articles/srep22032. The paper reveals the key advantages to a researcher that a ptychographic imaging approach can bring. The non-invasive properties of the technique were demonstrated by observing the maturation of primary hippocampal cultures into neurons with extensive neurite connectivity, together with the proliferation of microglia and astrocytes. Quantitative phase time-lapse movies of cells within the three populations were generated, which can be viewed alongside the publication. Crucially, since quantitative data was available on every single cell, motility metrics such as speed, meandering index and Euclidean distance could be calculated and these could be linked with changes in morphology, thus deepening understanding of the behaviour of individual cells over time. In this paper, (which used VL21, the Livecyte predecessor), cell tracking was carried out manually using Image J, but using Livecyte, powerful tracking software (patent pending) allows tracking of cells automatically rather than the manual approach employed here.
JR: Livecyte allows researchers to interleave fluorescence measurements during different drug treatments or across cell types within a single experiment. Why is this an important step forward?
TZ: Although there are clear advantages to a totally label free approach, the Livecyte system does have the ability to combine label-free and fluorescence protocols automatically and seamlessly, offering new experimental approaches. Protocols can be used which are predominantly label-free in their sampling frequency, but allow periodic tracking of fluorescently labelled components. This still reduces phototoxic effects, but also enables additional verification, if it is required. Label-free and labelled results can then be accurately correlated, avoiding inconsistencies that may be experienced if using different instrumentation for each. An example of where this could be useful is in reporter gene assays, or where functional identification of intracellular features is required. The flexibility afforded by this approach could be particularly useful to extend the time-courses of more fragile cells, while still minimizing their perturbation.
Tracy was speaking to Jack Rudd, Editor for Technology Networks.