Mettler-Toledo International Inc. (MTD) Earnings Call Transcript & Summary

Hello, and welcome to Mettler-Toledo and Maintenance webinar series. Today, we will be discussing good pipetting practices followed by micro volume measurement tips and tricks. My name is Alexandra Hellberg, and I'm the technical applications consultant for the West Coast. I've been with Mettler-Toledo for 5 years now, and I specialize in the UV/VIS spectroscopy portfolio.

Gustavo Chavarria, I'm the product manager at Rainin and specialized in liquid handling equipment. Here's how to get the most out of today's webinar. Engage and tune in. Don't worry if you need something. The webinar is being recorded. We'll make sure you have access to all webinar materials after the event. Your opinion accounts send us your questions and comments using the chat feature. Technical issues, reload your browser first to establish a new connection and communicate with us through the chat. Today, we are focusing on laboratory solutions, but don't forget that Mettler-Toledo offers a broad range of products that apply to the entire value chain, make it easy to streamline processes and has productivity and optimize costs.

Now we will begin the webinar presentation. We'll see you after for a live Q&A.

You may wonder why GPP or good pipetting practices. Well, the answer may not be that obvious, but even the subtle changes may affect your final dispenses and ultimately, your results. Let's take a close look at pipetting. We all know using an effective pipette will affect your protocols. No questions about that. But what about other factors? Are they critical? In general, there are 4 factors that will influence the delivery volume, the instrument, [indiscernible]; environmental factors, say, humidity and room temperature; type of sample and pipetting techniques. All of them will influence your results. What I was saying here. Well, calibrating your pipette is not enough. You can't control the nature of your samples or environmental factors, but you can control your techniques. In fact, techniques may play a bigger role than you think, and this is why we're here today to learn more about how to do better pipetting. And license protocols are run in almost every laboratory from simple research to clinical diagnostics. The plot on the left shows an ideal allies standard curve, sigmoidal, no major deviation in any of the under points but as we all know, that rarely happens. Science happens, right? And at some point, somehow, we end up with a plot that looks like the one on the right side. What happened? We always blame an expired reagent, the antibody is all miscalculated dilutions. At the end, it's always the reagents or the play reader. But have you ever thought that using the right pipetting technique could actually have prevented the situation. There are studies that actually have shown that pipetting technique could be a critical factor to consider. For example, bobrick's work published and concluded that an optimal pipetting technique is recommended to ensure the detection of weekly reactive antibodies. ELISA is just one example but there's been clear evidence that pipetting will affect any given protocol. The good news is you don't have to invest money in mastering the right technique. You need to know some do's and don'ts and when to use the right tools and techniques. First, let's take a quick refresher and the basics of pipetting. Most of your applications use air displacement pipettes or simply known as pipettes. There are 2 possible pipetting techniques you could use with these instruments: forward and reverse. The difference, forward is the technique always do in the lab. It's great when dealing with aqueous solutions. You press a plunger to the first stop, while you keep the tip inside the liquid, you slowly release plunger in order to start aspirating. The cycle continues as you press the plunger down to dispense the liquid all the way to the first half. The dispense is complete once you press the plunger to the second stop. And remember, this is important. You must press plunger to the second stop in order to dispense the expected volume. For reverse pipetting, which is recommended when dealing with viscous or dense samples, this cycle is a little different. If you notice, the cycle starts by pressing the plunger to the second stop. This allows you to pick up more sample at the beginning. Viscous samples tend to stick inside of the tip, the rest of the cycle continues similar to the forward pipetting technique. Now there are some techniques you could do with pipetting that will improve your results. We're not saying you must do every single one of them. You can pick and choose the techniques that are more convenient based on your protocol, the expected outcome or simplicity. Some techniques do require more practice than others, but each one of them can have a positive impact in your protocols. Let's are reviewing them with these examples. Perhaps this technique is the simplest to execute, yet not many people are aware. When setting up the desired volume with the micrometer always turns a micrometer 1/3 revolution above the desired volume. For example, if using a P10 to set at 2-microliter volume, then move the micrometer all the way to 2.5 approximately, then dial down to the desired volume, in this case, 2 microliters. This technique improves our accuracy by 0.5%, which in 2 microliters can be quite significant. We always recommend to dial passenger desire volume, 1/3 and then turn down to your final volume setting. Why is that? Well, the micrometer and the microballs have gears and those gears aligned inside of the pipette. When you dial up the gear can nest in front behind or top of each other. If you always dial down, they will always nest in the same configuration. This slide doesn't necessarily show you a technique per se. Rather, it illustrates the importance of choosing the right pipette for your application. Say you want a pipette 2-microliters into a UV spectrometer. In theory, you could use a P2, a P10 or P20 to do this task. The first page shows raining published technical specifications, systematic and random errors. When looking at the specs, look deeper both systematic and random errors are usually difficult to interpret in terms of our regular pipette user performance. At raining, we use accuracy or uncertainty a concept that allows you to evaluate your pipetting technique or the pipette performance by using 1 value instead of 2. A lower value means the dispense volume will be closer to the desired volumes. A large value means the variation will be larger. Table 2 shows theoretical uncertainty of these 3 pipetting using 4 repeated measurements. You can see the best pipette to dispense to micropipette is obviously P2. And you can see how using a P10 increases the uncertainty 4x. Why is that? Because for P2, you are using the pipette at the 100% nominal volume with 2 microliters is only 10% of the nominal volume for the P20. In general, the expected working nominal range for almost any pipette is 10 to 100%. Remember, never use the pipette for any volume that is out of range. The closer you are to the 10% higher chances to see larger uncertainties in the delivery volumes. The next technique is the angle when pipette and tip are immersed. We all have been there. We were told we need to hold the pipette particular to the surface to achieve the best accuracy. Is this true? Absolutely, but is it practical? Do we execute this every single time we pay pipette. The good news there is a practical working range to dispense liquids with better accuracy. As a recommendation, a [indiscernible] pipette between 0 degrees, which is the vertical and 20 degrees. The next slide deals with a technique that is a little more challenging than holding your pipette at the right angle. Where to place a tip relative to the surface of the liquid is not an easy task, especially when dealing with a small center rich tubes or opaque containers. Fit immersion depth can greatly influence the results. If the tip is not immersed enough in the liquid, there is a risk of sucking up air and not aspirating the right amount of volume that needs to be dispensed. If the tip is in merged too deep, liquid will rise into the tip because of the effect of air pressure. In this case, you could aspirate more sample volume than you desire. This slide shows recommended depth based on the pipette you are working with. For micro volumes, 1 or 2 millimeters should be good enough to get the best possible accuracy. This technique is easy to remember for micro volumes, 1 for 1 or 2 for 2, 2-millimeter or 2-microliters. As a general guideline, the pipette tip should just break the surface of the liquid sample. This technique is more of a recommendation rather than a technique, the aspiration rate. We have been there in the lab trying to rush or finish a plate, and you're trying to do it a quick by pipetting, you've also noticed that if you buy back too fast, you end up aspirating a volume that was not expected? Or you also noticed you sucked up air and end up with air pockets in your sample. Then you have to purchase the tip and repeat the process again. Not only you're wasting time, you're also compromising the integrity of the pipette. If you are not using filter tips, the liquid can splash into the shaft and damage the piston and the seals. You're also compromising your sample by introducing cross-contamination. So take a deep breath relax and aspire and dispense at a controlled steady pace. Avoid faster rates, is not going to finish your protocol faster. On the contrary, you're wasting time. Also, for those who are environmentally conscious, that tech is unlikely to be reused and you have to get a new fresh step to redo that pipetting spec. When dispensing liquids, you always have to do the touch off. That is touching the tip along the side of the vessel to complete the pipetting cycle. This can be done in 3 different ways: one, by touching the sidewalk or by placing a tip above the liquid surface or three, by dispensing the liquids into receiving liquid. Now this one is one of the techniques that confuse people the most, which technique is the correct one. The ideal is a touch off. During blowout slide along the sidewall of the tube while pulling up to ensure that all liquids have been spelled the touch of. But in general, there's no round technique. So ultimately, the answer is it depends. There's no wrong technique, just factors to consider that made dispensing the most appropriate. For example, a strong contamination of factor to consider can now use the tips, et cetera. However, there are things that you could do to improve your accuracy. If dispensing liquid keep the planter pressed at the second stop on the lid tip is completely removed from the sample, and this will be preventing an intentional aspiration. Progressing. These technique works, but very few people get to do this one. Why? Mostly because it's perceived as time consuming. Now keep in mind, it's never a bad idea to Pre-rinse your tips before dispensing. Why? Data shows that Pre-Rinsing tips twice or even once can greatly improve your accuracy up to 2% in the case of manual pipettes and 4% for electronic pipettes. So remember, for better accuracy aspirate the same liquid sample that you will be dispensing into the tip and dispense back into the reservoir or waste container. Repeat this ones or twice. Pre-Rinsing provides a uniform contact surface for all aliquots. In short, Pre-Rinsing does work. Hand warming can affect the results. During regular use, heat is transferred from your hand to the body of the pipette, prolonged handling of the pipette such as during micro lay work can result in a noticeable deviation impacted performance. prolonged handling of a pipette can cause a change in the delivery volume over time of as much as at 0.5%. This is primarily due to the expansion of the air column inside of the tip from hand warming. So the recommendation is that after 10 minutes of use, you should hang up your pipette on the stand to allow the heat to dissipate. This is also a good economic practice to allow your hands to rest and stretch. Keep in mind, the rest period for economic reasons need only to be between 2 and 3 minutes. We want to spend a couple of minutes on this slide. So far, we've been talking about techniques that can help your accuracy with aqueous Solutions and have limit of implications for other liquids. But as we know, we work with numerous samples that are more complex or challenging that pipetting water. From these examples like proteins to volatile solvents such as ethanol or acrylonitrile, there are specific techniques that can greatly improve your accuracy when working with challenging liquids. For this purpose of the webinar, we can classify the liquids into subgroups based on some physical characteristics. Most of the time, you have to work with samples that tend to create foam, better volatile, that are viscous or have low surface tensions. For all of these samples, you can do 2 things: use diversified technique or use low retention tips. I'll get back to this point in a second. Pre-Rinsing is highly recommended for all liquids, but viscous and using white [indiscernible] will help to dispense all liquids except volatile. As a reminder, forward technique is highly accurate when dealing with aqueous solutions only. Now low retention tips are not a technique person but there are a great alternative to improve your accuracy when you are doing the forward technique. The table shows that percent rotation of the liquid is far less when using a regular standard tip. The figure shows a graphical representation of what I'm describing. For example, pipetting 50% of isopropanol improves 11x just by using a low retention tip when compared with standard tips. And remember, deep selection matters. Always use the tip recommended by the pipette manufacturer and check the tip ceiling before pipetting. In summary, all techniques are valuable by selection and execution of these recommended tools depends on the ultimate goal of your protocol. Some techniques are more difficult than others or consuming and you don't have to execute them all every single time. My suggestion is start with the easy ones, then micrometer, immersion angle and aspiration rate. And make them a habit for daily pipetting and use the more difficult ones when protocol requires special attention and excellent accuracy. And now Alexandra would like to talk to you about some exciting instrumentation that can help you with your research for protocols.

Thank you, Gustavo. As I mentioned in the beginning, my name is Alexandra, and I will be taking you through some slides regarding successful micro volume measurements. First, we'll start with a micro volume spectroscopy overview where we'll cover some basics about micro volume measurements, the most common applications and then review the Mettler-Toledo Vivid portfolio. As most life science laboratories have an arsenal pipettes, they usually have some type of spectrophotometer as well. There are a few reasons this type of instrument is so crucial to life science research. The first being that it's able to measure concentrated biological samples in an undiluted manner. So only a few microliters of sample are needed to cover a wide range of concentrations for nucleic acids and proteins. The fact that an additional dilution step is not required for these types of measurements, really helps us to eliminate those potential points of error. We're instead able to pipette our sample directly on to the platform and the instrument will automatically select the appropriate path length and calculate the results that we ask for. A few of the most common applications performed at a micro volume spectrophotometer include nucleic acid bacteria and protein analysis. Both the purity and the concentration of nucleic acids play an important role in life science research because these values help us to identify and therefore, prevent failures in downstream experiments such as PCR. In order to determine nucleic acid concentration, the absorbents at 260 nanometers is used. Further calculations can then be added to specified double-stranded or single-stranded DNA as well as RNA. Not only is the concentration of these nucleic acids as important value to note, but the purity of these samples is as well. A purity check will help us to gauge potential contamination within the sample as well as the success of the extraction process. The A260/A280 ratio gives us a level of protein contamination within the sample while the ratio A260/A230 directs us towards other contaminations such as EDTA, urea, phenol, et cetera. Another common application where micro volume spectrophotometers can be beneficial is with cell density, which we refer to as OD600. By looking at the absorbent levels at 600 nanometers, we're able to monitor the growth of bacteria cultures. And this is most accurately done by creating a standard curve and then measuring your sample against it to identify the unknown cell density. Now moving on to proteins. A micro volume UTVs measurement is used to perform a quick check to identify the protein concentration of a specific sample. And this can be done by simply measuring the absorbents at 280 nanometers. A simple measurement at 280 nanometers, however, is pretty vague and because it measures mainly triptofan and phenylalanine it might in accurately estimate the concentration value when we're measuring complex protein mixtures or pure proteins. So in order to avoid this an indirect measurement using a quantification curve will help us to provide more accurate results. Widely expected procedure for indirect protein quantification involve the use of BSA as a standard for the Bradford assay. So taking a little deeper into the Bradford assay, as Gustavo mentioned earlier, I have outlined here the process of how protein concentration is determined with this assay. Firstly, proteins form a complex with Coomassie brilliant blue dye. The die will actually change color from a reddish-brown to a bright blue based on acidity. Bradford assay contains this die in a esthetic solution, which means it starts off as that reddish brown color. The protein binding will lead to a change in color from the reg brown to a bright blue, which can then be measured at 595 nanometers, which is where blue falls in the visible spectrum. So essentially, we create a series of standards from Redish Brown and color all the way up to bright blue. And then the instrument is able to use this series of values to identify the protein concentration of our unknown sample. And as a side note, our best practice recommendation is to create this calibration curve with the same proteins that are contained in your sample. Now that we've covered the basics of micro volume measurements and some of the key applications that this type of instrument is used for, I wanted to highlight the difference between pipetting a few microliters of sample onto a platform and using a few milliliters of sample inside a cuvette for these type of measurements. This is referred to as macro volume spectrophotometry. The inner workings of the instrument are nearly identical with the main difference being that with the micro volume model, the light passes up through the sample droplet reflects off of a mirror, passes back through the sample and then continues through the fiber optics towards the reflection grading and then is ultimately detected by the CCD sensor. With the macro volume, however, the light simply is collimated through a lens and pass through the cuvette containing the sample and then follows the same path to be read by the CCD sensor. I thought this was interesting to point out because the older model macro volume instruments still out there today are enormous, and they take up almost an entire bench whereas the micro volume instruments have always been more compact. With this new technology, however, both instruments are finally the same size with Mettler-Toledo. This is a quick overview of our UV/VIS portfolio and all of the models actually sit on a standard 8x11 piece of paper. So you can see our micro volume option in the front there, which we call the UV5Nano and it has both micro volume and cuvette or macro volume capability. The other models the UV5Bio, the UV5 and the UV7 are each suited for different types of applications and industry-specific methods come preloaded onto each of those instruments. Each model in our portfolio is able to perform a scan of the entire spectrum in 2 seconds and also has the ability to easily export data to a USB drive. The UV5Nano specifically comes preprogrammed with life science method shortcuts and also a free rein in pipette. Here's a nice visual of the life science bundle, which includes the UV5Nano that can accommodate both micro volume and cuvette based measurements, the 10-microliter LTS, RAININ pipette as well as [indiscernible] RTL 10 tips. So now that we've covered the basics of micro volume measurements as well as the Mettler-Toledo UV/VIS portfolio, I'll now go into some tips and tricks for you to improve your current micro volume analysis. First, we will do a quick refresher of proper liquid handling, which Gustavo has gone into more detail for us. But I will touch on pipetting technique, dispensing technique and sample application. So as Gustavo already mentioned, we want to be aware of immersion depth when transferring a sample from a small vessel onto the micro volume platform of the UV instrument. This is something we want to be aware of because if the tip is immersed too far, air can become compressed in the tip and too much liquid can be aspirated. And if it's not immersed far enough, air bubbles can be drawn in, resulting in inaccurate volumes being dispensed. I wanted to touch on this point again because correct tip emersion depth can actually improve measurement accuracy by up to 5%. Another refresher from Gustavo's presentation is maintaining a vertical angle during aspiration. Keeping the angle as close to vertical as possible can improve measurement accuracy by up to 2.5%. Now moving on to some dispensing techniques specific to micro volume measurements. And following these steps will help to yield perfectly formed droplets on the platform and therefore, repeatable and accurate results. So first, you want to pipette 3 microliters of sample into the tip, place your hand on top of the instrument to study yourself and rest the tip on the center of the platform. Then carefully dispense the sample and immediately take note of the droplet formed on the platform. If the droplet does not bead and form a nice semi-circle or if you can see air bubbles trapped within it, simply bought that sample with the Kimwipes and repeat the previous steps because these will not yield successful results. Once a proper droplet has been pipetted onto the platform, simply closed the micro-volume arm choose the appropriate shortcut on the terminal and start the measurement. 2 path length are available on the UV5Nano, both 1 millimeter and 0.1 millimeter. The longer path length, which is 1 millimeter, can accommodate 3 to 5 microliters of sample while the short path length, which is 0.1 millimeter, can handle 1 to 3 microliters. Here, we have a visual representation of the importance of using the pipetting technique, which leads to a nice round bubble of sample on the measuring platform. Despite the fact that we are talking about a few microliters of sample, it's pretty apparent after you've pipette it on to the platform, whether or not a bubble has formed or has fallen flat. The reason this bubble is so crucial to the accuracy and repeatability of micro volume measurements is because as we saw in the earlier schematic drawing, the light has to pass through the sample from the bottom, reflect off of the mirror and then pass back through the sample in order to be analyzed. After the sample is pipetted and the arm is closed on top of the sample, the instrument will lock into place at the desired path length, which is what we are looking at with these bottom 2 drives. So on the right, we can see that after the arm is closed since the bubble didn't form properly, there's a portion of air between the sample and that top mirror. This means that when the light is passing up through the sample, it's also passing through air before it's reflected off of the mirror. And this unfortunately leads to incorrect measurement values because the instrument is taking into account the transmission of sample as well as that air rather than just measuring the sample itself. Now that we've covered some good pipetting practices as it relates to micro volume measurements, we will talk a little bit about cleaning. So which parts of the spectrophotometer need to be cleaned, some do's and don'ts and also how to perform a cleanliness check. We also have a nice QR code at the bottom right-hand of the screen, what you can scan with your phones and will be led to a cleaning routine video that can be very helpful. So beginning with which parts of the spectrophotometer need to be cleaned. The micro volume platform, which is circled at the bottom there, as well as the upper mirror component at the top are both crucial parts of the optical measuring system. If you're more familiar with cuvette-based spectroscopy, a good way to think about these 2 components on the screen here is considering them to be the front and back walls of the cuvette. So this means that both the micro volume platform and that upper mirror need to be spotless in order to achieve accurate and repeatable results. Now starting on the right hand of the screen with materials and cleaning agents that we recommend. Kimwipes or other brands of lint free optical tissues are a great tool to clean the platform in between blagging and simpling and also in between sample measurements. And air blower can then be used to remove any remaining just lint or even liquid from the platform in the mirror. Acceptable cleaning agents include DI water, spectroscopy grade isopropanol or ethanol, disconnect solution or diluted HCL or pepsin when working with protein specifically. As for do's and don'ts when it comes to cleaning, starting with the do's, be sure to wear gloves to avoid contaminating the platform or the mirrors with fingerprints. Also be sure to clean and dry all components of the measuring system in between each run and let DI water or a mild cleaning solutions sit on the platform for a few minutes on a regular basis as a deep cleaning step. And moving on to the don'ts. Be sure to avoid cleaning agents other than what we have listed here as it could damage the measuring system. Be careful not to push and spin the mirror on cleaning and instead dab firmly, but still gently. And lastly, don't allow any type of solution, especially samples or stronger cleaning agents to sit on the platform for extended periods of time because these can degrade components and ultimately affect the accuracy of sample readings. The last thing you want to avoid when it comes to micro volume UV/VIS measurements is overfilling. Be careful to never pipette at more than 10 microliters of sample or blank onto the platform as this can damage the instrument over time. Excess liquid can lead to corrosion of the arm and actually reduce the accuracy of the system itself. Now that we know the proper way to clean and use our micro volume UV/VIS instruments, how do we know that we actually cleaned it well enough. So after you've cleaned the platform with DI water and wiped clean twice, you will blank as you usually would. Then instead of adding a sample, you will actually add your blank again and run it as a sample. The absorbent should end up near 0.01A, and if it's close to this value, you've successfully cleaned the surfaces. If the value is high, though, the cleaning process was not sufficient and needs to be repeated before running any samples. So to this point, we know what a micro volume UVs instrument is, what it's used for, how to properly pipette that samples onto the platform, how to clean the platform. And then lastly, we will cover how to ensure repeatable results. So here's a zoomed-in diagram of the sample being compressed by the arm and locked into the desired path length. It's important to note that each sample is pipetted onto the platform should only be analyzed by the instrument once. That is because when the arm is lifted and lowered repeatedly, a layer of air can get trapped between the sample and that upper mirror. If air bubbles are being introduced each time the arm is lifted and then lowered the intent of running a sample in duplicate or triplicate will be negative because those bubbles will actually be changing their results after each analysis. So to avoid this, be sure to use a new sample for every repeated measurement that you'd like to perform. To conclude my segment, I've summed up the content we discussed today into 4 main points: First, is that precision and accuracy are essential for nucleic acid and protein concentration, a sample loss can be critical. These types of samples require an instrument that can handle limited volumes available as is without needing to be diluted, which is where our micro volume spectrophotometer comes into play. Secondly, good pipetting practices and a cleaning routine need to be followed in order to avoid cross-contamination and skewed results. Thirdly, sample droplet and sample size need to be closely monitored during the pipetting process in order to get the most accurate and repeatable results. And lastly, Mettler-Toledo and Rainin are here to support your life science needs with a complete solution when it comes to small volume sample analysis. And now I will hand things back over to Gustavo to tie everything together with something called Smart Check. So we talked about the importance of sample size when it comes to using a micro volume spectrophotometer, so he will discuss a simple way to validate your pipettes prior to use.

Earlier to this webinar, we suggested that P2 is the best instrument to deliver samples up to 2 microliters, but what if you don't have it, what if -- and this is very likely that you only have a P10 or P20. How can you know your pipette or your technique will deliver the best 2 microliters? Remember, pipetting should be considered as assistant not as individual components. As a reminder, pipetting has 3 components: instruments, tip and techniques. It's not enough to calibrate or service pipettes. It is also important to test your techniques as well. Now a quick check of your pipettes is traditionally done with an analytical balance. What's the problem and consuming. You have to 0 the balance, you have to pipette into a vessel, wait until the reading stable and collect the data. Then you have to use software to analyze the data and calculate the mean and standard deviation, which are the systematic and random errors. Imagine doing this every single time to every pipette, it will never end. What will you say if I told you there's a more efficient, faster alternative that could tell if your pipette is working properly, or if your technique is appropriate without using a balance or running the calculation. Mettler-Toledo and Rainin would like to introduce 2 Smart Check. The first pipette verification system in the market that can check both pipettes or your pipetting techniques. Smart Check only needs 4 dispensers. Performs all necessary calculations. Give you a definitely past failed response, so no need of extra calculations, and it works with any pipetted brand. Now we would like to play a short video to show you how Smart Check works. [Presentation]

Thank you so much for joining us today. We hope you enjoyed and learn some new techniques to implement into your workflow. So hello there. Are there any questions?

If there's no questions, we can go ahead and conclude the webinar. Definitely feel free to check out our survey link, and we also have the presenters contact info if you think of anything after the webinar. Thanks again.