Let’s get started then. Thanks for your patience. I’m really happy that you could join us today for the Optura webinar. This webinar will be recorded and it will be made available following the live events today. So in this webinar, we will go through the theory behind how the palm takes a measurement, some product specific details. We will then watch a demonstration of the palm where we will show what’s in the box and how to get started with the device and how it generates a biomass calibration. And then we will take a look at applications. Finally, at the end of the webinar, we will have a Q and A session. So please ask questions in the box on your screen. Firstly, let’s introduce the Optura Palm. This is a handheld portable device which is designed to measure total biomass in shake flasks and other vessels. The device gives it on the spot reading by pressing a vessel up to the measurement face on the palm. Not only does this device free up time by providing a quick insight into your biomass by giving a reading in five to seven seconds, it also minimizes the risk of contamination due to its contactless nature where the flask does not even need to be opened to take a measurement. The palm works with a variety of vessel types including glass and plastic shake flasks, falcon tubes, duram bottles, and any other labware which is generally optically clear. The device also works with different cell types. There are some generic biomass calibrations which are preloaded to the device, but you can generate your own calibrations to suit your needs, which we will show you later. So the Optura palm measures bio reflectance, or in other terms, backscatter. The principle is the same as the Optura SPY for real time monitoring, but in a portable form. In simple terms, a single wavelength is shone directly out of the sensor into the media. This is then reflected off the cells present in the media. That signal is then received back into the sensor via the source detectors. As the cell concentration increases, the amount of reflectance also increases as there are more cells to reflect off, and therefore, the detector will detect a higher amount of reflectance, which correlates with increase in biomass. The raw reflectance measurement given out by this measure instrument is nanowatts. This is the absolute unit of a measurement that the instrument is calibrated to. And although nanowatt might not be familiar to the user, this region can be easily converted to more familiar units like grams per liter, offline o d six hundred, things like that. So our bio reflectance sensors use an optimized wavelength, which is in the near infrared range. And specific to the palm, this is thirteen ten nanometers. This path length has been chosen for multiple reasons. The first reason is that when using the light source wavelength close to near infrared, absorbance in water is strong. Now water is always present in fermentations and cell culture in the form of liquid media, which means that the laser always has good penetration. At this wavelength, the colored media constituents also have minimal impact on the reflectance measured. By using thirteen ten nanometers, the path length required to take a measurement is also reduced to a distance which is only two and a half centimeters of unimpeded space in front of the measurement face. That equates to roughly two hundred microliters of a sample in area, which is about the width of a fifty mil falcon tube. So this means that we can essentially measure in quite small spaces. It is important to understand the differences between transmit and space measurements and reflectance measurements, especially with the palm, as often people will be looking to transition away from a benchtop spectrophotometer, which is transmittance based. So a transmitting based measurement will pass light through a sample and measure how much light transmits through the sample, which is then received on the other side. The higher the cell density, the less light that is passed through. This method is prone to saturation of the sample, which either requires a different wavelength to be used or for an offline measurement, They ABER responsive to biomass across a whole measurement range. In this section, we will have a little look at a pre-recorded demonstration of the palm so that you can see it being used. This is a demonstration of the Optura palm, a handheld contactless device for monitoring total biomass in a range of vessels including shake flasks. Firstly, here is how the palm will arrive, in a protective hand carry case where the sleeve is removable. To open the case, you need to open the two latches. This is the quick start guide. This simple guide tells you how to get started with the device. We recommend that you read this before using the palm. Next is the Optura palm device itself. This is the verification block to intermittently verify that the device is working as expected. And this is the USB cable for both data transfer and charging the device. The Optura palm works by emitting a near infrared laser from the measurement face, which is located at the end of the device. When this laser interacts with biomass within a flask, the light is reflected back into the device. The higher the biomass, the higher the return of light. This reading is by default measured in nanowatts as a calibrated unit. However, this can be converted to a more familiar biomass measurement type, such as cells per mil, grams per liter, or optical density. To take a reading with the device, power it up using the power button on the screen. Pressing the cal button allows you to scroll through any biomass calibrations on the device. This includes the factory default nanowatts and some preloaded biomass library readings. Select the biomass calibration that you would like to use. The palm measurement is initiated when the two pins present on the measurement face are both engaged against a rigid vessel wall. Ensure that the flask is clean and the cells are in suspension. You may need to tip the vessel to ensure that there is enough liquid in front of the sensor. This should be around two and a half centimeters. Simply press the palm against the vessel, hold the palm and flask still for five to seven seconds, and your readings will be complete. Your measurement will appear at the bottom of the screen next to the measurement number. One thousand readings can be stored at any time. Now that you’ve seen how the palm takes a reading, let’s walk through the verification procedure. To use this block, remove the protective lid to expose the surface of the block. This should be inspected before use for any fingerprints, dust, or debris. This can be wiped with a lint free lens wipe. If you need to use IPA, spray that on the wipe first rather than spray in the block. The same advice should be taken to clean the measurement face off the palm. To take a verification reading, cycle through the cal button to find verification nanowatt. Hold the block against the measurement face, engage the pins by pressing firmly, and you will hear the tones along with the screen prompt, which indicates the measurement is in progress. Your measurement should lie between the min and max tolerances printed here. When taking a reading with the palm, if the verification block or flask is unstable or the pins disengage prematurely during the measurement, a negative tone will sound, along with a red light and a warning message. If this happens, just check your alignment and try again. If you would like to generate your own biomass calibration, you can do this by either using the palm to monitor your flasks that you already have going, or you can do this by taking readings in a dilution series. Here, I have multiple flasks containing different concentrations of yeast. I already know the offline values and have those written down. A biomass calibration should be made for a particular cell type, flask, and media type. If you change up one of these parameters, it is advised that a new biomass calibration is made. Firstly, we must show the device cell free media in a flask. This is to introduce the zero value to the calibration so that subsequent regions are biomass only. You should take this reading in the factory default nanowatt calibration. It is recommended that regions are taken in triplicate when generating a biomass cal. I will start with taking three regions in cell free media. Then I will move across the dilution series, noting down which reading number corresponds to each flask. We would recommend using at least three different cell concentrations spanning across the range of biomass that you use for a better biomass calibration. This software allows the user to configure their device, download, save, and delete measurements, and also to generate a biomass calibration. When your Palm device is connected to your computer using the USB cable, open up palm connect. Click the search for devices button and find the palm that you would like to connect to. Now the palm is connected, you will see all of the previous measurements taken using the device. Here, you can save all of your measurements on the device, delete them, or generate a biomass calibration. To generate a biomass calibration, select the measurements that you wish to use and select new calibration. The selected measurements will show in this table. Simply check your offline values, find the right reading, and type your offline values into the reference column. Here are my offline readings for cell free media. So I will type zero in the reference column. If you aren’t happy with one of the points, just delete the reference value and it will be left out of the calibration. When you are happy with the fit, you can now save your calibration. Give it a name within sixteen characters, and that will show up on the palm device itself. Click save to the device. Now, your brand new biomass calibration is on your device and ready to use. I hope that you enjoyed that video. So now that you have seen how the device works, we will take a look at some of the data with the device using various cell types and flasks. Now let’s have a little look at what the data looks like across a dilution series, starting with microbial cells. Here is a plot which is based on a E. Coli culture. So this plot reflects a dilution experiment where E. Coli cells were grown in LB media in clean glass shake flasks. The cells were spun down to achieve a very high cell concentration and then diluted with spent media to test the concentration range. So as you can see, a very good correlation of zero point nine nine was observed between the palm nanowatt signal and offline OD measured at OD six hundred on a benchtop spectrophotometer. So the concentration range was roughly zero point five OD up to seventeen OD before the device saturated. So when the device saturates, the device can no longer take a stable reading, and you will receive consistent unstable reading values. Another example of an E. Coli dilution series can be found here, but for the Thompson UltraYield flasks. These flasks aren’t completely transparent like some other plastic flasks. However, the near infrared laser can still measure through these types of vessels. The only thing that you will lose is some of the resolution of the device. Here you’ll see a really good correlation between the readings taken using the palm on this type of flask and an offline OD at OD six hundred. Continuing with microbial cells, here are some plots of yeast, both Saccharomyces cerevisiae and Pichaea pastoris. Again, these regions were taken in glass shake flasks in LB media and another dilution series was performed. You will see good correlation across the board with different yeast strains and different offline methods including offline OD and total cell density, which was measured in cells per mil using a hemecytometer. Although these measurements were taken using the palm with a glass shake flask, plastic shake flasks also perform roughly the same. Now to mammalian cells. So in this case, CHO s freestyle cells were monitored in CHO s freestyle media in one hundred and twenty five mil plastic shake flasks. A good correlation was observed, and the use of the palm meant that the shake flasks didn’t need to be opened to get a reading, which maintains sterility. The minimum concentration detection limit across the devices is three million cells per mil. Mammalian cell cultures give off a lower reflectance return than microbial cells. This is apparent in the biomass calibration slopes. It’s believed that this is due to the cell wall composition as microbial cells have cell walls, whilst mammalian cells lack a rigid cell wall. You will see this clearly when looking at slope values of the preset biomass calibrations available on the device. You should always expect the slope value from a correlation with mammalian cells to be higher. In this section, we will go through some of the more technical details on the palm, and these will be handy for you to know and understand. So on the palm device itself, you will have a biomass calibration library. So these are generic calibrations aimed to show you what could be achieved with the device. So these should not be taken as absolute as they were generated in house at ABBA, And these calibrations are based on our cells that we use. So the calibrations that are preloaded consist of nanowatts, so that’s the raw measurement, verification nanowatts, so that’s for the verification mode. These two cannot be deleted. We also have E. Coli at o d six hundred, Saccharomyces cerevisiae at grams per liter, and CHO in cells per mil. There is space for sixteen calibrations in total in the device, and the first two, as I mentioned, can’t be deleted. The others can be deleted in the software to make room for your own calibrations. So on this slide, we can see some of the concentration range limits. These do differ between different cell types as the cells look very different to the device and they reflect the light back into the device quite differently. So for E. Coli, we can go down to zero point five to fifteen point five o d six hundred, give or take. Really depends on your media you’re using, your flask you’re using, lots of different little factors there. We also have Saccharomyces cerevisiae, which is around zero point three to six grams per liter or zero point eight to thirty five OD and OD six hundred. Pachaea is around one to thirty five OD six hundred. And we also have CHO cells, which is three million cells up to one point nine times ten to the power of eight. So since the palm device is used with labware and the vessel needs to be tipped to get the right penetration depth as you’ve seen in the video, this table describes the minimum fill volumes that are required to take a measurement for your most standard lab items, including your shake flasks, Duran bottles, tea flasks, and fifty mil falcon tubes. Any vessel types that have a lot of striations in the from the plastic, which is from the plastic molding procedure, should be avoided, and any really scratched glassware should also be avoided as these can impact the reflectance. If you have any questions about vessel types that you can’t see in this table, you can get into contact with us, and we’ll give you some advice. So when using the palm, it is also important to understand the effects of wall thickness on the measurement as this can impact the detection limit of the device. The way this limits the device is that the zero value will be increased. For instance, if I was taking a reading in YPD media only in different wall thicknesses, so for us in this instance, polycarbonate was used, the reflectance return in the media will increase with an increased wall thickness. Beyond four millimeters, this starts to lower the upper detection limits of the device. So as you can see here, at two millimeter wall thickness, you can achieve a maximum cell concentration of up to thirty five OD before the device starts to saturate. At a five millimeter wall thickness, the upper limit is reduced to around twenty one o d. And at a ten millimeter wall thickness, this reduces to thirteen point five o d. Whilst it’s really hard to predict the effects with different vessel types and materials and cell type combinations, this is good to keep in mind, especially if you’re looking to measure through thicker vessel wall materials. So thank you very much for attending the webinar, and I hope you found it very informative and useful. We’ll be sending a follow-up email with the webinar recording, and we will go into the q and a session now. I will just be reviewing your questions, but please continue to submit them. And if we don’t get round to answering them, we’ll make sure to send you an email after.