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Pipette Tips

David Suzuki Love the wild bees

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The Definitive Source For Lab Products, News And Developments

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March/April 2016

How tips influence results

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Exploring between diabetes the linkand exercise


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CONteNts 10

ON THE MOVE

DON’t FORGet tHe WiLD Bees by daVid SuZuKi

by hErMiOnE wilSOn

Public attention has been focused on domesticated honeybees, but the bee crisis also affects hundreds of wild bee species equally vital to the ecosystem.

ulriKE gaSt, iOan gligOr,

The research team at the Physical Activity and Diabetes Laboratory (PADL) is studying the links between exercise and the treatment of diabetes.

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staNDaRDs

Do the flip! to learn about emerging diabetes treatments.

CaNaDiaN news 6 Guest editorial 9

laura KOch, and rOnJa KubaSch

The pipette tips you use in the lab can have a big impact on your results.

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March/aPril 2016

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LaB ware 18

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teCH watch 20 MOMeNts in time 21

tiP OF tHe iCeBeRG

by MuriEl art, VincEnt dufEy,

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suzuki matters www.labbusinessmag.com

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n March I had the opportunity to volunteer in my son’s grade three classroom. I was there to participate in an exciting program run every year called, Scientists in the School (SiS). If you’re unfamiliar with the program, the website says, “We’re the fieldtrip that comes to you!” and that’s pretty accurate. The idea is to make science fun and relevant, allowing students to become scientists in their own classroom by participating in investigative activities which follow the curriculum. In my case, scientist Jane Beakbane ran a workshop called, “Soil: It’s too important to be treated like dirt!”, a subject which the children had already been introduced to in their own classroom learning. A retired scientist, Beakbane became involved in the program years ago while looking for something to do in her spare time. She’s never looked back. She now crisscrosses the region running workshops on a variety of topics. It keeps her busy and she enjoys staying involved with science and working with children. On that day, Beakbane divided the classroom into four different workstations/experiments, each run by a parent volunteer. Children were able to cycle through each station and then take part in a discussion and Q&A with the teacher and herself at the end. Beakbane brought in different types of soil, as well as earthworms, and as the children participated in the experiments and discussions and touched the materials, they learned about soil composition, erosion and the role earthworms play. I found it interesting and of course, hands-on learning is much more fun and memorable than simply reading something in a handout. I found the experience very rewarding both as a parent and as editor of this magazine. It was wonderful to spend time in the classroom with my son, meeting his classmates, teacher, as well as other parents. I appreciated being part of their day and watching all those little light bulbs go on as their understanding of all the concepts came together by the end of the workshop. Whether the children went home and told their families what they learned around the dinner table or whether they use the knowledge in their gardens this spring, or simply take out a related book from the library, who knows where it will lead them next. And that’s the beauty of learning.

Theresa Rogers EXECUTIVE EDITOR

Publisher of lab buSinESS Magazine biO buSinESS Magazine Printed in canada

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Canadian NeWs IBM Partners with Hamilton Health Sciences

UBC Celebrates Women in the STEM World The University of British Columbia marked International Women’s Day this year by hosting an event that examined female leadership in science, engineering and medicine. UBC organizers invited notable women who are leaders in their field to share their experiences and the lessons they have learned. Among them was Nadine Caron, the first aboriginal woman to graduate from the UBC Faculty of Medicine and Maria Klawe, President of top U.S. engineering, science and mathematics school Harvey Mudd College.

Science Fair Launched in Montreal

Dino Trevisani, President of IBM Canada, and Rob MacIsaac, President and CEO of Hamilton Health Sciences, interact with Pepper the Robot.

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BM and Hamilton Health Sciences (HHS) recently announced they would join forces to help area hospital clinicians, researchers, academics and entrepreneurs accelerate the development and commercialization of new healthcare innovations. The two companies plan to establish a new centre in downtown Hamilton that will serve as a physical and virtual collaboration space for healthcare innovation. IBM is contributing access to an array of its Watson cognitive and analytics software, expertise in cloud computing and high-performance computing infrastructure, and a network of global collaborators. HHS, with its cadre of more than 1,500 principal investigators and research staff, provides practical industry expertise and a “realworld” test environment. The centre, which will open its doors this fall, is expected to put Hamilton on The centre, which will open its the map as a hub of healthcare innovation doors this fall, is expected to put in Canada. Hamilton on the map as a hub of “Hamilton is among the top cities in healthcare innovation in Canada. Canada for health research innovation,” says Hamilton Mayor Fred Eisenberger. “This partnership is proof of the growing platform we have here to launch new ideas and turn them into new products that improve health care and create the jobs of tomorrow.” In two of the first projects, IBM and HHS will test how IBM Watson cognitive and analytics capabilities can be applied to HHS’ existing decision-support database, to provide insight into how patients use the healthcare system; they will also explore introducing a mobile component to add functionality and scalability to HHS’s early warning system, which electronically monitors a patient’s vital signs for subtle changes indicative of a worsening condition or pending medical event.

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March/april 2016 LAB BUSINESS

The 2016 Hydro-Québec Science Fair launched in March in Montreal. Young people aged six to 20 years old participated in different levels of competition, from the regional and Quebec provincial finals, to the Canada-Wide Science Fair. Outstanding exhibitors at the regional finals will get the chance to participate in the Super Exposciences Hydro-Québec, Quebec final 2016 in April. Forty young exhibitors will also get the chance to represent the province at the Canada-Wide Science Fair in Montreal in May.

CFI Funds Research into the Alberta Oil Sands Two University of Calgary researchers have received funding from the Canada Foundation for Innovation’s (CFI) John R. Evans Leaders Fund. Geography professors Scott Jasechko and Brent Else colead a research project studying the carbon and water cycles from the oil sands of Alberta to the Arctic Ocean. CFI-funded equipment will be used to study groundwater movement in the oil sands, water vapour transport in the Arctic, carbon dioxide absorption by Arctic Seas, Arctic Ocean carbon cycling, and carbon transport via flowing groundwater.


Worldwide NeWs Vancouver-based Biomedical Inks Deal with China

Materials Researchers Recognized The Materials Research Society (MRS) announced its 2016 Fellows at the MRS Spring Meeting in Phoenix, Arizona. Among the 14 MRS Fellows recognized for their sustained and distinguished contributions to the advancement of materials research were researchers from Stanford University, The University of Cambridge, and the U.S. Naval Research Laboratory. MRS is an international organization that promotes the advancement of interdisciplinary materials research and technology to improve the quality of life.

Distek Inc. Turns 40 Carolyn Cross, CEO of Ondine Biomedical (centre), signs joint venture agreement with Yanan Zhao (far right), with Zhang Wanming, Director of a Free Trade Zone in Henan Province of China (left) in attendance.

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ancouver-based Ondine Biomedical has inked a deal with China to bring its photodisinfection technology to the Chinese market. Carolyn Cross, Chair of the Board and CEO of Ondine Biomedical, recently signed a joint venture agreement with Yanan Zhao, CEO of Zhengzhou Zhenghe Medical Devices, which will begin with the introduction of two non-antibiotic technologies, Periowave (for treatment of oral infections) and MRSAid (for the prevention of surgical site infections, a leading cause of hospital acquired infections). In use across Canada, Ondine’s technology provides rapid antimicrobial efficacy to prevent and eliminate infections without encouraging the formation and spread of antibiotic resistance. Ondine’s photodisinfection therapy for hospitals debuted in the Pre-Surgical Decolonization Project in Vancouver General Hospital, which received the 2013 In use across Canada, Ondine’s Innovation Award of Excellence technology provides rapid from the International Consortium antimicrobial efficacy to prevent for Prevention & Infection Control and eliminate infections without (ICPIC) in Geneva. encouraging the formation and The Pre-Surgical Decolonization spread of antibiotic resistance. Project, which reduced infection rates by 39 per cent, was a 12-month non-antibiotic pilot program involving more than 5,000 patients who were treated with MRSAid photodisinfection therapy prior to a major surgery. Since implementing MRSAid nasal decolonization to pre- surgical patients, VGH now enjoys one of the lowest surgical site infection rates in Canada. “Antibiotic resistance has been declared to be a global threat to humanity by key organizations such as the World Health Organization (WHO) and the National Health Service (NHS) in the UK,” Cross says. “We are pleased to bring our technology to China and are looking forward to working with our Chinese partners to see our life saving photodisinfection technology in Chinese healthcare facilities.”

Distek Inc. was founded by Gerald Brinker, a sales engineer in New York City with a B.S. in Chemical Engineering from McGill University in Quebec. In 1972, Brinker formed a manufacturers’ representative firm for analytical instrumentation called Brinker Instruments. As the sole proprietor, Brinker was responsible for launching new products for many laboratory instrument manufactures. In 1976, he renamed the company Distek and had it incorporated. Distek is now a leading manufacturer of pharmaceutical laboratory testing instruments.

North American Institutions Among Top-Ranked Research Innovators In March, Reuters published its Top 25 Global Innovators list, a ranking of government research institutions around the world. The ranking factors include patent filings, published academic articles, performance and more. Seven North American institutions landed on the list, including the U.S. Department of Health & Human Services (No. 4), The U.S. Department of Energy (No. 8), and the National Research Council of Canada (No. 14). Europe dominated the list, with nine out of 25 ranked institutions. Asia came in second with eight.

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suzuki MatteRs

BY DAVID SUZUKI WITH CONTRIBUTIONS FROM JODE ROBERTS

Love bees

– especially the wild ones!

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Dr. David Suzuki is a scientist, broadcaster, author, and co-founder of the David Suzuki Foundation. Jode Roberts is communications strategist and urban beekeeper. Learn more at www.davidsuzuki.org.

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any environmental campaigns over the past 50 years have aimed at getting people to care for imperiled species in wild, far-off places. The focus in Canada has often been on large, photogenic, culturally important animals, with bonus points for campaigns that include alliteration, bumper sticker-friendly slogans and plush toys. This has been a sensible and often successful strategy. Over the past few years smaller, charismatic critters closer to home have buzzed into the spotlight: bees. About a decade ago, beekeepers in Europe and North America started noticing serious declines in honeybee populations. Bees have lost much of their natural habitat to urbanization and industrial agriculture and face increased stress from climate change-related drought and severe winters. These threats, coupled with the global spread of diseases and pests and a dramatic increase in the use of agricultural pesticides like neonicotinoids, have resulted in unprecedented losses for beekeepers. (Because bees and other insects provide ecological services like pollination, it makes no sense to declare war against all just to eliminate or control the few nuisances.) The honeybee decline has been big news partly because they make delicious honey, but more importantly because they’re pollinators. About three-quarters of flowering plants and more than a third of food crops worldwide depend on pollinators – from bees and butterflies to hummingbirds and bats. As a result, governments across the globe are developing strategies to protect them, including Ontario with its recently proposed Pollinator Health Action Plan. Public attention in Canada has largely focused on domesticated European honeybees, but research indicates the honeybee crisis is part of a wider problem affecting hundreds of lesser-known but crucially important wild bee species. Of about 800 wild bee species in Canada, more than 90 per cent have a “solitary” lifestyle rather

March/april 2016 LAB BUSINESS

than living in large, social colonies. Two-thirds of these are ground-nesters, including bumblebees, mining and digger bees that make nests in soil and under leaves and rocks. The rest are cavity-nesters like mason and carpenter bees that burrow in hollow stems, twigs and logs. Although honeybees get the headlines and most of the credit for pollinating flowers and crops, studies show that wild bees can be two or three times better at pollination, and some, like mason bees, can be up to 80 times more effective. The good news is that the honeybee crisis has galvanized interest in all pollinators, inspiring thousands of groups and citizens worldwide to establish new spaces for them, from wild bee hotels and rooftop honeybee hives to pollinator gardens in parks and schoolyards. As our communities grow, pollinator habitat is fragmented into increasingly disconnected patches that disrupt natural pathways, making the potential of connected networks of habitat within cities especially fascinating. Oslo’s Bumblebee Highway, Seattle’s Pollinator Pathway and Hamilton’s Pollinator Paradise are all great local initiatives. Establishing an urban pollinator corridor is also at the heart of the David Suzuki Foundation’s Homegrown National Park Project, which since 2013 has created more than 50 pollinator-friendly patches along the path of a creek now buried beneath Toronto – from small guerrilla plantings to a network of flower-filled canoe planters in schools, cafés, churches, parks, and yards. This spring, the Foundation will launch the Great Canadian Butterflyway Project, to inspire beefriendly urban innovations and neighbourhoodscale pollinator corridors across the country. Through videos, tips and other resources, the project will profile projects nationwide that are bringing nature home, one pollinator-friendly planting at a time. You can become part of the growing movement to protect pollinators. Head to the library (or check out davidsuzuki.org/pollinators) to research the amazing diversity of wild bees and other pollinators in your community. While you’re there, learn what flowers and shrubs best support those species, and what might work in your yard or on your balcony. Then check out what local groups are up to. Want to show wild bees some love? Create a sanctuary in your yard or garden by leaving a sunny patch of bare soil for ground-nesters. Add some pithy stems, sticks and wood debris for cavitynesters. And be sure not to disturb the nests over winter. Will the buzz generated by media stories and pun-filled campaigns save the bees? Only time will tell. In the meantime, we can all help by making bees welcome in our yards and neighbourhoods. LB


Guest eDitORiaL

the big things matter BUT NEVER AS MUCH AS THE LITTLE THINGS

BY

Robert Price

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don’t entirely understand what people mean when they talk about the economy. Every election a politician will drop the magic word as if it means something. They’ll say something like, “I’m for the economy.” Applause will follow. There are other magic words. Terror. Globalization. The Environment. Innovation. Each one is an abstract noun we need to “fix.” It’s no wonder so few people vote. When politics devolves into abstraction, why bother paying attention, let alone voting? It’s easier to care when a specific issue comes to the fore. Like ending home mail delivery. Then politics is real. No doubt, the big stuff in life matters. But we can’t talk about the big stuff if we treat these concepts as if they are the issues themselves. The economy is a just an idea. It’s not a thing but the product of a million little decisions. Same with globalization. It isn’t a mystical force bending our economy toward foreign and corporate powers. It is, in fact, the outcome of entirely local decisionmaking. We chose to open ourselves to trade and capital. It wasn’t imposed on us. The same is true with Big Data. Reading about it in the news, you’d think Big Data was a force of nature, like Godzilla. If we take off our futurist glasses, we see it for what it is: the outcome of a series of decisions we’ve made personally and through public policy to acquiesce our privacy rights to governments and business. And the same problem creeps up when we speak about “Science.” We make a mistake when we treat the discipline as a monolithic actor on the public stage, at

war with Religion and fighting for Reason against the forces of the Irrational. I’m guilty of this kind of thinking; sometimes it feels as if great forces are at work. It can seem as if there is a battle for civilization happening in the clouds. Maybe there is. But if it is battle for the future, it’s a ground battle. And it’s local. Because the person who makes each little decision has power over the rest of us. We saw this fight over the last decade when Harper’s Conservatives prohibited publicly funded scientists from speaking publicly about their work. That decision wasn’t an attack on Science, nor was it an abstract contest of ideologies. It was instead a political decision – censor public scientists to protect the government’s “brand.” It was all a ground game. Scientists are right to demand policy instruments that protect the integrity of their scientific work. Recently, the Professional Institute of the Public Service of Canada and Evidence for Democracy, a lobby group, asked the new government “to clarify the rules for government scientists, protect the integrity of their research and make it harder for future governments to muzzle scientists.” The government should take the advice of these lobbyists and enact provisions that maintain the integrity of scientific findings. Government should decide policy based on sound evidence. It shouldn’t have right to tilt, erase or game evidence to suit its decisions. Enacting a few small changes to protocol – for instance, permitting a government scientist to answer a reporter’s phone call – will make a huge difference to what we call Democracy. We’ll have the chance to be more informed on the issues when scientists can speak. The economy, terror, globalization and just about every “-ism” are abstractions. And there’s really nothing we can do with them. What we can do is remain vigilant to how we let others – government and business – nip and tuck the rules. It is small decisions – whether tucked into an omnibus bill or sent as a directive through the public affairs department – that change the world. LB Robert Price is former Managing Editor of this publication. Follow him @pricerobertg.

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ON THE RESEARCHERS AT THE UNIVERSITY OF ALBERTA EXPLORE THE LINK BETWEEN DIABETES AND EXERCISE STORY BY

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fOr StEinbacK, thE linK tO diabEtES in hiS rESEarch iS hEart diSEaSE, which iS, accOrding tO bOulÉ, thE nuMbEr OnE KillEr Of PEOPlE with diabEtES.

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March/april 2016 LAB BUSINESS

Hermione Wilson

alking by the Physical Activity and Diabetes Laboratory (PADL) at the University of Alberta, you might mistake it for a fitness centre. A large part of the lab is exactly that, a private research gym where Dr. Normand Boulé and his colleagues, Drs. Margie Davenport and Craig Steinback, run their training studies. The fitness centre serves as an exercise and physiology lab, and is divided into different subsections. The main section is dedicated to the exercise and fitness testing, while the other sections are for drawing blood, observing exercise capacity and oxygen consumption, and taking other measurements of participants before, during and after physical activity. Two smaller rooms at the back of the lab are where the more experimental work takes place, where Boulé and his colleagues look at more advanced measures of a participant’s cardiovascular function and how the body responded to different tests.

Since 2010, the PADL has been a part of the University of Alberta’s Alberta Diabetes Institute (ADI) and has made its home on the first floor of the Li Ka Shing Centre for Health Research Innovation. The institute is made up of researchers from different faculties at the university, such as Nutrition (one floor above the PADL), Pharmacology, Public Health, Immunology, and the faculty from which Boulé, Steinback and Davenport hail, Physical Activity and Recreation. “Our faculty has been part of the [Alberta Diabetes] Institute for a long, long time,” Boulé says. “It’s just more recently that this building became available and we worked hard with the faculty and the Alberta Diabetes Institute, as well as the Alberta Diabetes Foundation, to put this lab together.” Although Boulé, Davenport and Steinback have physical activity as a common thread in their research work, each of them specializes in three very diverse areas. Boulé looks at how exercise, in conjunction with medication and diet, affects glucose control in people with diabetes. Davenport studies the impact of exercise and sedentary behaviour on maternal and fetal health, while Steinback is mostly concerned with how the brain controls blood pressure and communicates with blood vessels. The three researchers have their own unique research focuses, but they do find many overlaps in their work where diabetes is concerned. For example, Davenport has in previous studies looked at blood sugar control in women who had gestational diabetes and how exercise could be used as an adjunct therapy in standard prenatal care, along with diet and insulin. For Steinback, the link to diabetes in his research is heart disease, which is, according to Boulé, the number one killer of people with diabetes. “Glucose and insulin are not regulated as well [in people with diabetes] and can have a confounding effect on the cardiovascular and autonomic systems,” Boulé says. Steinback employs a technique called microneurography to study how the body communicates with blood vessels and


Lab PROFiLe

regulates blood pressure. Using a device called a Nerve Traffic Analyzer, Steinback can record tiny bioelectrical signals that correspond with bursts of activity and elevated levels of stress, and amplifies them. “We take basically [something] like acupuncture needles and we can place those to record the activity of nerves in the body,” he says. “We can record how the body responds to stress, or your fight or flight response, and that response directly speaks to blood vessels, and that in turn is what controls your blood pressure.” Using data acquisition software, Steinback and his team can merge these

signals with other measurements of blood pressure and blood flow, recorded using ultrasound technology, in order to get a more complete picture of what is going on. Steinback is also interested in how individuals respond to activities, environments or health conditions that limit their oxygen intake. In the lab he uses gas mixtures to change the composition of the air a participant is breathing through a mouthpiece or face mask in order to simulate breathing at high altitudes. Then he observes how the body responds to the stress. Steinback is planning a research expedition to the Mount Everest region this fall to study the effects of the low oxygen environment of the mountains on the human body. He plans to study the local population of sherpas and porters and how their bodies have adapted to their unique environment, as well as doing tests on himself and the group of 30 researchers he plans to bring with him. “We’re going to be our own subjects,” Steinback says. Both Boulé and Davenport have

Top: A fitness centre serves as an exercise and physiology lab at the Physical Activity and Diabetes Laboratory. Photo Credit: Craig Steinback Above: Normand Boulé analyzes a study participant’s oxygen consumption and carbon dioxide production during an exercise test. Photo Credit: Alberta Diabetes Institute

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Above: Three researchers share lab space and often collaborate on studies such as how blood pressure is regulated during pregnancy. Photo Credit: Zoltan Kenwell Right: A post-doctoral student monitors a participant’s blood pressure while she uses a recumbent cycle ergometer. Photo Credit: Alberta Diabetes Institute collaborated with Steinback on a number of projects. Davenport uses the same ultrasound and microneurography equipment when she and Steinback are doing work together in the area of cardiovascular regulation. “We’re looking at how blood pressure is regulated during pregnancy, as well as during complicated pregnancies, so women who have high blood pressure or have diabetes during pregnancy, or are at risk for developing high blood pressure,” Davenport says. “We’re looking at some of the underlying mechanisms that might cause this to happen during pregnancy.” Steinback and Davenport plan to collaborate in the near future on the impact of sedentary behaviour on blood glucose control and overall health. “When they both arrived, I could see links more directly with [Davenport] and her gestational diabetes,” Boulé says. “The word diabetes is something I understand a little bit more than some of the stuff [Steinback] does, but I think from a mechanistic perspective, some of the collaborations between [Steinback] and I weren’t expected but have developed as well.” In the course of their research and the studies that they have conducted both together and separately, Boulé, Davenport and Steinback have uncovered some interesting things about the effects of exercise on human physiology. They have learned, after several studies looking into health outcomes for combining the drug Metformin and exercise, that the two don’t necessarily augment the effect of

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daVEnPOrt iS currEntly wOrKing On a SEt Of PrEgnancy and EXErciSE guidElinES, and bOulÉ waS rEcEntly brOught On tO wOrK On thE PhySical actiVity SEctiOn Of a nEw EditiOn Of thE canadian diabEtES aSSOciatiOn guidElinES. the other. Metformin is one of the most commonly prescribed medications for diabetes. “The combination of the two was no better than exercise alone or Metformin alone,” Boulé says. “We don’t have a good happy ending story yet where we can say, ‘This is how you should best try to combine them to have better outcomes,’ but we’re working on that as well right now, how to better combine exercise with these other mainstream interventions.” They are also learning interesting things about resistance training and whether higher intensity exercises are beneficial not just for athletes and young healthy people, but people with diabetes as well. On the opposite side of the spectrum, Davenport and Steinback have discovered, during the course of their research into sedentary behaviour, that low energy activities that involve sitting or reclining are independent predictors of long-term cardiovascular or metabolic diseases. “The idea of not exercising enough or exercising too little is completely separate from sitting too much. They’re very distinct ideas, so we’re looking at both of them,” Davenport says. She is

particularly interested in the effects of sitting too much on cardiovascular and metabolic function during pregnancy, about which little is known. The three researchers of PADL have helped shape the changing protocols for diabetes and pregnancy exercise guidelines. Davenport is currently working on a set of pregnancy and exercise guidelines, and Boulé was recently brought on to work on the physical activity section of a new edition of the Canadian Diabetes Association guidelines. “Some of our initial successes had a lot to do with really improving the standards of the evidence behind exercise in Type 2 diabetes,” Boulé says. “At the time, a lot of the initial studies were kind of small, underpowered, and a bit varying results, so we did a lot to synthesize the evidence and come out with some really good quality evidence behind the role of exercise.” Ultimately the work Boulé and his colleagues are doing at PADL will have a lasting impact not only on those struggling with diabetes, but also for anyone – young, old, relatively healthy or living with illness – who wants to live a better, healthier life. LB


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The Tip of the Iceberg: How Pipette tips influence Results STORY BY

Muriel Art, Vincent Dufey, Ulrike Gast, Ioan Gligor, Laura Koch, and Ronja Kubasch

ABSTRACT Pipettes are tools widely used in the lab and usually purchased with care. However, as pipette tips are only consumables, they are not usually selected with quality in mind. The standard ISO 8655:2002 recommends using pipette and tips from the same supplier. This study, including tips from 15 different manufacturers, proves that a pipetting system working perfectly with a certain tip exceeds the permissible error tolerances when a tip from a different manufacturer is used. Furthermore, we found that the calibration method influences the performance of the pipetting system: It is significant whether the calibration is done with or without tip change for each measured volume. Autoclaving impacts

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March/april 2016 LAB BUSINESS

the tip dimension as well as the calibration result especially with small volumes. INTRODUCTION Within the scientific community, a rising number of experiments published cannot be reproduced by other groups. Incorrect pipette handling (e.g. holding the pipette at an angle during liquid aspiration) may be one reason for this. A second source of error often not taken seriously is plastics. Consumables may lead to problems with analysis results, e.g. due to leachables, as well as incorrect pipetting volumes. This may result in non-reproducible data if experiments are repeated by other groups using other consumables. Some problems with pipette tips are obvious like:

> Tips have to be pushed powerfully onto the pipette cone in order to achieve efficient tip fit. > Banana-shaped tips make it difficult to fill a plate with multichannel pipettes. > Pipetting of volumes below 1 ΟL on a solid surface is impossible because the liquid drop sticks to the outside of the tip. In the same way that only the tip of an iceberg can be seen above water level, a number of other problems with pipette tips rather stay unknown. One example is decreased pipette accuracy when using tips not recommended by the pipette supplier. The latter often stays unnoticed since problems with analysis results are usually linked to reagents, method and pipette but not to the pipette’s consumable.


application NOte Moreover, calibrations are understood as “checking the pipette” instead of “checking the system”. Thus, instead of checking the tips utilized in the lab only the tips recommended by the manufacturer are used for calibration. The ISO 8655-21 defines the pipette and tip to be a system. It stipulates extra calibration for the use of other manufacturer’s tips. But why does this standard put so much focus on a product that is to be discarded after use? This paper shows the influence of tips on the pipetting result. It explains the origin of such influences and what to look for when purchasing tips not recommended by the pipette manufacturer. Basically, the main influencing factors are design/shape, production quality and material. These APPLICATION NOTE I No. 354 I Page 4 factors do not only influence the single pipetting result, they also have methodical consequences like varying Results and Discussion results when calibrating with one/several tips or changing by of autoclaving. Influence of tips on theresults performance the pipetting system The aim of this paper is to picture the The >pipette and tip< system was perfectly within the error iceberg underneath the level:theto limits when Eppendorf tips were used.water It was outside specifications when used with tips from other manufacturers. generate understanding why the pipette As shown in fig. 1, the systematic error was exceeded in form a µL system withC, both 4and cases its with atip volume of 1,000 (manufacturers E, K, N)

components having equal influence on pipetting results. RESULTS AND DISCUSSION Influence of tips on the performance of the pipetting system The pipette and tip system was perfectly within the error limits when proprietary tips were used. It was outside the specifications when used with tips from other manufacturers. As shown in Fig. 1, the systematic error was exceeded in 4 cases with a volume of 1,000 μL (manufacturers C, E, K, N) and in 5 cases with a test volume of 1 μL (manufacturers A, E, F, H, M). With three of those 1,000 μL tips, the test volume not only exceeded the manufacturer specifications but also the wider maximum permissible systematic error as stated by the ISO 8655:2002 standard1. In contrast, the random error was increased noticeably but stayed within permissible tolerances. If all calibration results are combined, a total number of eight from 15 manufacturers exceeded the not only exceeded the manufacturer specifications but also specifications. However, it error cannot the wider maximum permissible systematic as statedbe byassumed the ISO 8655:2002 standard [1]. In contrast, the random that all tips from a manufacturer error was increased noticeably but stayed within permissible are affected if with one tip the calibration tolerances.

and in 5 cases with a test volume of 1 µL (manufacturers A, E, F, H, M). With three of those 1,000 µL tips, the test volume 10 µL

1 µL

0.5

2

0.4

Random error in %

Random error in %

0.45 1.5

1

0.5

0.35 0.3 0.25 0.2 0.15 0.1 0.05

0

-4

-2

0

2

4

6

8

0 -1.5

10

-1

-0.5

0

1

1.5

1000 µL 3.5

3

3

2.5

2.5

Random error in %

Random error in %

100 µL 3.5

2 1.5 1

2 1.5 1 0.5

0.5 0

0.5

Systematic error in %

Systematic error in %

0

-5

-4

-3

result exceeded the permissible error limits. For example, the 10 μL tip from manufacturer K performs within error tolerances whilst the 1,000 μL tip from manufacturer K exceeds the pipette manufacturer’s and ISO 8655 specifications. Calibration results were found to be independent of the pipette manufacturer and were reconfirmed by calibrating with a pipette from another manufacturer (data not shown). This corresponds to the requirements of standard ISO 86551. All pipette users should be aware of the fact that when using tips not delivered by the pipette supplier, the manufacturer’s declaration or certificate of conformity does not apply. The ISO 8655-21 clearly states the pipette and tip to be a system and recommends using those tips supplied by the pipette manufacturer. In case this is not possible, the ISO 8655 part 21 describes that the user has to perform a conformity test first with tips from the pipette supplier. If the pipette passed the test, a second calibration with tips not supplied by the pipette manufacturer has to be performed. Coming back to the calibration results and taking a closer look at the volumes being most impaired: We found a clear difference between 10 μL and 1,000 μL results. With 1,000 μL tips, the nominal volume (1,000 μL) was found to be most affected whereas with 10 μL tips, the 10 % of nominal volume (1 μL) was found to exceed the technicalspecifications. From this finding, it can be deduced that the violation of systematic error limits has different reasons with 10 μL and 1,000 μL tips. Those main influencing factors are described in the following.

-2

-1

0

Systematic error in %

-5

-4

-3

-2

-1

0

Systematic error in %

Figure 1: Calibration results using 10 µL tips and 1,000 µL tips from different manufacturers. The colored area shows the span of the maximum permissible errors

stated for the Eppendorf Xplorer pipettes. All data points within the colored area are within the specifications. Figure 1: Calibration results using 10 μL tips and 1,000 μL tips from different manufacturers. The coloured area shows the span of the maximum permissible errors stated for the Eppendorf Xplorer pipettes. All data points within the If all calibration combined, a total number of exceeded the permissible error limits. For example, the 10 µL coloured area areresults withinarethe specifications.

8 from 15 manufacturers exceeded the specifications. However, it cannot be assumed that all tips from a manufacturer are affected if with one tip the calibration result

tip from manufacturer K performs within error tolerances whilst the 1,000 µL tip from manufacturer K exceeds the pipette manufacturer’s and ISO 8655 specifications.

Tip design and its influence on pipetting results The dimensional measurements of length and inner diameter showed that some 1,000 μL competitor tips were longer than proprietary tips with the same inner diameter. These longer tips failed the calibration. In order to explain these findings, it has to be taken into account that pipettes in general are adjusted to a certain air cushion size and filling height of liquid within the tip.

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15


Tab. 2: Results for MEA test of Eppendorf 50-1,000 µL Biopur and 2-200 µL Biopur tips. The test is passed if the test item has no effect on growth and development to at least 80 %.

application NOte Longer, bigger or slimmer tips lead to an increased total size of air cushion and a different filling level5. If the dead air volume increases, the pipetting volume decreases. Additionally, an increased filling height (e.g. by slim and long geometry of tip) results in an increased hydrostatic pressure which has to be compensated and also leads to a decreased volume and higher systematic error6. Our data show that with 1,000 μL tips the shape-related influencing factors play an immense role. This effect is especially distinct at nominal volume because with 1,000 μL the biggest possible “weight” has to be moved by the air cushion. We see that the tips dimensions do not solely explain the calibration results. Other factors like wetting and quality-related issues, for example, perfection of tip orifice, come into play. Quality of tip orifice The zone where the liquid leaves the tip during dispensing is very important for the accuracy of results. At this part of the tip, the drop cut-off occurs. Any imperfection of geometry or shape, e.g. by production failures, leads to water retention. This especially plays a role with small volumes. A poor drop cut-off may not only impair the pipetting result but can also make it impossible to dispense small volumes: sometimes a drop leaves the tip, sometimes it doesn’t. A good tip has a front phase with a defined wall thickness and surface structure in order to ensure a good drop cut-off. Production tolerances need to be very tight. A poor orifice is not perfectly round or has walls of differing thickness. Liquid drops become deflected to the outside of the thinner wall. Furthermore, it shows “molding flashes” or thin “flashes” where liquid may be retained. In addition, “lying flashes” influence the diameter of the orifice and again lead to liquid retention. From the production view, these flaws occur mainly if poor tools are used especially in combination with a non-optimized injection molding process. Generally, it is recommended to use tips which are made of a non-wetting plastic material with a flawless smooth orifice5.

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March/april 2016 LAB BUSINESS

Conclusion Within the scientific community, there are a rising number of studies which cannot be reproduced by other groups. One possible reason may be that the influencing factors of pipette tips are not taken into account – just like recognizing

only the tip of an iceberg. We showed within this study that tips from different suppliers can alter the pipetting result and its reproducibility. Thereby, different influencing factors become effective:

Air cushion size Tip shape

Filling level of liquid Tip fit on cone

Factors decide on:

Tip orifice

> Accuracy of system > Reproducibility of results

Water retention Tip material

> Methodical influences on results

Autoclaving behavior

a) tip change/no tip change within

Leachables

analysis b) autoclaving

Perfection of orifice Production quality

Perfection of sealing rim Tip-to-tip quality

Some non-system providers offer tables showing on which pipettes certain tips fit. Our results show that a tip fitting onto a pipette cone does not say anything about the pipetWater retention onour inner surface ting result. Furthermore, results underline that it does not make much sense to use a “universal”“wetting” tip if the pipette Water retention, so-called is does not become calibrated (and, if needed, adjusted). Tips not only influenced by the tip’s orifice but are an important component of the system and they are optimized pipette they are produced for. Accordingly, also byfor itsthe material and inner surface. the ISO 8655 [1] regards the pipette and tip to be a system If the inner surface is uneven or the tip and recommends using tips and pipette from the same

is made of an unfavourable material, liquid will be retained on the surface inside the tip. However, a completely smooth inner surface is not the only solution for minimizing water retaining effects. The combination of material composition and surface structure decides on the water retention. Tips are generally made from a plastic called polypropylene (PP). However, PP is not PP. Each tip manufacturer has its own secret formula for the PP it uses to produce pipette tips. From cooking we know that the secret behind a good cake is good ingredients. This also applies to the PP for tips. The mixture of ingredients determines the water repellent characteristics. PP in general is hydrophobic. This can be observed by pipetting drops of water onto different surfaces (e.g. glass and plastic). The rounder the drop, the higher the surface energy and the less the wetting effect. Within this study, we found differences in volume of retained liquid between different tip manufacturers. Since pipetting errors sum up, like holding angle of pipette during uptake or different temperatures of pipette and liquid, high water retention can cause a pipetting system to be out of specifications. Tip fit – Influence of sealing rim on creation of an air-tight system It is a basic requirement of a tip to fit

manufacturer. Our results are evidence that this requirement is meaningful and we strongly recommend naming the tip used within publications and to calibrate (if needed: adjust) the pipettes if other manufacturer tips have to be used.

TIP

As a final overview, tab. 3 displays the results of all experiments done within this study. It shows that Eppendorf tips Sterilization processes performed by keep the promise of highest quality performing best overall.

users are usually not monitored. Thus the sterility is questionable. The benefit of purchasing sterile products from a manufacturer is an assured sterility.

tightly onto a pipette cone in order to generate an air-tight system. The term “tip fit” applies in two different contexts: firstly, does the tip physically fit onto the pipette cone? Secondly, does this tip fit provide a tight sealing? The physical fit of a tip onto a cone is mainly influenced by the design and shape of its contact zone with the pipette cone. The tip diameter is not the only factor. The connection between tip and pipette cone needs to be tight enough to prevent air passing through. If the tip fit is not tight, the system does not aspirate enough liquid and may leak. In the worst case, the pipette is obviously dripping but incomplete tightness is often not recognized during daily lab work. It was shown that the pipetting error increases by up to -0.6 % systematic error and 0.8 % random error with the generic tips used7. In order to ensure a tight fit on the pipette cone, most tips are equipped with a sealing rim. The position as well as the quality of this sealing rim is important for the tip fit. If the sealing rim is positioned too low, the cone may not reach it. If the sealing rim is too thick, it takes


application NOte TIP Additives easing production process (plasticizers, biocides, slip agents) are known to disturb biological assays. Look for manufacturers that avoid their use. Additives needed for product characteristics (e.g. not becoming brittle) should be at a minimum.

high forces to fit the tip onto the pipette cone. This has a negative effect on ergonomics and pipetting comfort. The consistent quality of the sealing rim itself is important for the tightness of the system. Its thickness must not vary significantly from tip to tip. Variations in the quality of the sealing rim mainly originate from three different production failures: 1) the tools used to produce a tip are not equally shaped or are maintained with too infrequently, 2) the injection molding process is not

balanced enough to ensure that the sealing rim is perfectly shaped (e.g. not enough plastics injected), 3) application of excessive production tolerances in quality control in order to save money in production. CONCLUSION Within the scientific community, there are a rising number of studies which cannot be reproduced by other groups. One possible reason may be that the influencing factors of pipette tips are not taken into account – just like recognizing only the tip of an iceberg. We showed within this study that tips from different suppliers can alter the pipetting results and their reproducibility. Thereby, different influencing factors become effective. Some non-system providers offer tables showing which pipettes certain tips fit. Our results show that a tip fitting onto a pipette cone does not say anything about the pipetting result. Furthermore,

References 1 ISO 8655:2002 parts 1 – 6: Piston-operated volumetric apparatus. www.iso.org 2 Eppendorf SOP: Standard operating procedure for manual dispensing tools. www.eppendorf.com 3 Eppendorf Xplorer® Operating Manual. www.eppendorf.com 4 ISO 10993-12:2012: Biological evaluation of medical devices. Part 12: Sample preparation and reference materials. www.iso.org 5 Lochner KH, Ballweg T, Fahrenkrog HH. Untersuchungen zur Meßungenauigkeit von Kolbenhubpipetten mit Luftpolster. J Lab Med 1996; 20 (7/8): 430-440 6 Salje G. Internal communication 7 Carle AB, Rodrigues G, Rumery D. Best Practices for the Use of Micropipettes. Poster. www.artel-usa.com 8 Wenk E, Lustgarten JA. Technology of Manually Operated Sampler Pipettes. Clin Chem. 1974, 20(3): 320-323 9 Knaide T, Albert KJ. Quantifying the Impact of Pipette Tip Type Using Dual Dye Ratiometric Technology. ARTEL Application Note 2011; www.artel-usa.com 10 Grzeskowiak R, Gerke N. Leachables: Minimizing the influence of plastic consumables on the laboratory workflows. Eppendorf White Paper No. 26. www.eppendorf.com 11 Olivieri A, Degenhardt OS, McDonald GR, Narang D, Paulsen IM, Kozuska JL, Holt A. On the disruption of biochemical and biological assays by chemicals leaching from disposable laboratory plasticware. Can J Physiol Pharmacol 2012; 90(6):697-703 12 Watson J, Greenough EB, Leet JE, Ford MJ, Drexler DM, Becastro JV, Herbst JJ, Chatterjee M, Banks M. Extraction, identification, and functional characterization of a bioactive substance from automated compoundhandling plastic tips. J. Biomol Screen 2009; 14(5): 566-72

our results underline that it does not make much sense to use a “universal” tip if the pipette does not become calibrated (and, if needed, adjusted). Tips are an important component of the system and they are optimized for the pipette they are produced for. Accordingly, the ISO 86551 regards the pipette and tip to be a system and recommends using tips and pipette from the same manufacturer. Our results are evidence that this requirement is meaningful and we strongly recommend naming the tip used within publications and to calibrate (if needed: adjust) the pipettes if other manufacturer tips have to be used. LB Excerpt reprinted with permission. For more information, visit www.eppendorf.com/eptips.

List OF aDveRtiseRs & WeBsites Mandel page 2 ................................................. www.mandel.ca caledon laboratory chemicals page 4 .....................................www.caledonlabs.com look good feel better page 13 ................................... lookgoodfeelbetter.ca Mottlab inc page 19 .......................................... www.mottlab.com Vwr page 22 .............................................................. vwr.com

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17


Lab WaRe SPEctrOScOPy SOftwarE haS POwErful nEtwOrKing cOMPatibility

Metrohm USA’s new Vision Air spectroscopy software for Vis-NIR instruments has an intuitive user interface for fast measurement and straightforward data interpretation. Its operation focuses on sample analysis parameters and trends, not spectra, increasing the information density from NIR measurements. Using an integrated networking system, the Vision Air software guarantees reliable measurements across multiple facilities and multiple operators by ensuring that configurations and methods are updated simultaneously. www.metrohm.com

lab cOat OffErS flaME and chEMical PrOtEctiOn

SuPErcritical EXtractOr uSES cO2 inStEad Of liQuid SOlVEnt

The new pilot scale supercritical extractor from Supercritical Fluid Technologies is designed to simplify extraction of botanicals, herbs, spices and other natural products. Instead of using traditional liquid solvent, the extractor uses a safe, efficient supercritical carbon dioxide fluid that has possesses many of the same characteristics with the added benefit of high diffusivity and pressure tunable solvency power. Additionally, no residual solvents are left behind in the extract or remaining biomass. There is no need to perform a distillation step since, upon dropping the pressure, the CO2 becomes a gas, leaving the extracted material in its pure, natural state. www.supercriticalfluids.com

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March/april 2016 LAB BUSINESS

cEntrifugE can handlE largE VOluMES

The Thermo Scientific Sorvall BIOS 16 centrifuge has an increased capacity of 16 l of cell culture product per run. Higher capacity rotors – four swinging bucket rotors ranging from 6 x 1000 ml to 8 x 2000 ml - makes working with large volumes easier. With its auto-door and auto-lid functions, and centritouch interface, the bioprocessing centrifuge is user-friendly and efficient. In addition, it has an accumulated centrifugal effect function that automatically adjusts run time to account for and eliminate acceleration and deceleration variations that could otherwise lead to incomplete cell pelleting. www.thermofisher.com

Workrite Uniform Company’s new lab coat is the first of its kind to include both flame-resistant properties and chemical –splash protection. The fabric of the Workrite lab coat provides protection against short-duration thermal incidents and emergency exposure to flame while resisting the penetration and wicking of inadvertent liquid chemical splashes. While providing this protection, the lab coat is also designed for comfort and usability, with a concealed snap front closure, black lay-down collar to indicate chemical-splash protection and three pockets, the lower two of which have snap front closures. www.workrite.com


Lab WARE Pulsed Flame Photometric Detector can Detect over 26 Analytes

Xylem’s new OI Analytical 5383 Pulsed Flame Photometric Detector (PFPD) is ideal for petrochemical, environmental lab, and food and beverage applications. The highly selective detector features improvements to electronics and signal processing, and easy-to-use software. The PFPD gives chemists the ability to analyze low levels of sulfur, phosphorus, and 26 other analytes of interest. The detector is also ideal for detecting organophosphate pesticides. www.xylemanalytics.com

Low Volume Samples Go a Long Way with Small Scale

PAC’s Herzog OptiFlash Small Scale uses very low volume samples of only 2 or 4 ml to accurately detect flashpoints for petroleum products, biodiesels, solvents, chemicals, paints and varnishes, food and beverages while complying to leading global standards such as ASTM D3828, ISO 3679 and ASTMD7236 (Ramp Method). The OptiFlash has an intuitive user interface and built-in automation, and can detect flashpoints from -30 C to 300 C. www.paclp.com

OEM Pump an Option for Surgical Ablation

The RXMD DriveSure panel-mount OEM pump from Watson-Marlow Fluid Technology Group was designed for use by medical device specialists serving the surgical ablation market. The OEM pump features DriveSure brushless DC gear motors with fully integrated speed controls, a tube holder to ensure that the tube locates in the right position every time the safety lid is closed, and a mechanism that improves flow accuracy. The pumps’ DriveSure gear motors are highly adaptable and directly accept market-standard analogue control signals. www.watson-marlow.com


tech WatCH

Pipettes r-VErtEX PiPEttE iS EaSy On thE handS

RV Instruments’ R-Vertex series of pipettes are designed for ease of use. They have a soft finger grip of thermoplastic, a large four-digit display panel, and are colour-coded for easy identification. A built-in, streamlined tip ejector facilitates easy tip ejection and access to narrow necked bottles and tubes. The shape of the pipette’s high-quality plastic body is designed to lessen fatigue during repeated use and its stainless steel piston ensures the longevity of the device. www.rvinstruments.com

tranSfErPEttE PiPEttE EaSy tO taKE aPart

BrandTech Scientific’s Transferpette S pipette has thumb-tip volume selection and a short range of motion that reduces the risk of repetitive strain injury. Both the single and multichannel pipette models are autoclavable at 121 C without disassembly for convenience and protection from contamination. Transferpette eight- and 12-channel pipettes feature individually removable tip cones for easy cleaning and seal replacement, which comes in handy if faults are discovered during calibration, as only the individual channel would need service. www.brandtech.com

nOthing lEft tO chancE with nEw SartOriuS PiPEttE

The Tacta mechanical pipette features new Sartorius Optiject levered tip ejection technology which enables controlled and smooth tip ejection with minimum force. The Optiload feature involves spring-loaded tip cones in both single and multi-channel models, which ensures tip loading with perfect sealing. The Tacta also includes the new Sartorius Optilock system which provides flexible volume adjustment and locking functionalities and prevents accidental volume changes during pipetting. Tacta pipettes can be autoclaved without disassembly and are highly resistant to exposure to UV and chemicals. www.sartorius.com

ElEctrOnic MultichannEl PiPEttE rEducES Strain

Eppendorf’s new Multipette was developed in line with its Physio Care Concept. In order to reduce the risk of repetitive strain injuries, the multichannel pipette has an electric motor-driven dispensing function and buttons positioned to accommodate natural hand movements. To reduce eye strain, the Multipette has a large easy-to-read colour display with clear arrangement of all adjustable parameters. The pipette includes the option to save often used settings as favourites, as well as a new multiple aspiration function and a display of unknown volumes with immediate dispensing in desired partial volumes. www.eppendorf.com

20

March/april 2016 LAB BUSINESS


Moments in tiMe

tHe

Miracle OF INSULIN D

iabetes was basically a death sentence before two Canadian researchers discovered insulin in 1921. Dr. Frederick Banting and his research assistant Charles Best were testing the theory that the pancreatic juices in diabetes patient were harmful to the secretion of the pancreatic islets. In their lab at the University of Toronto, Banting and Best surgically restricting the flow of digestive juices from the pancreas to the intestines of a healthy dog, then removed the organ, ground it up and filtered out a substance they named “isletin”. They injected the isletin into a diabetic dog and observed that its blood glucose level dropped. They were able to keep the dog healthy and symptom-free with a few injections a day. Eventually Banting and Best moved on to bigger animals like cows, renamed their extract “insulin” and began human testing. The two scientists were awarded the Nobel Prize in Physiology or Medicine in 1923 for their discovery, the same year medical firm Eli Lilly began producing enough insulin to supply the entire North American continent. LB

www.nobelprize.org/educational/medicine/insulin/discovery-insulin.html

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21


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LAB Business March April 2016  
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