 Okay, so good morning, good afternoon, good evening, whatever time you're watching this again. Welcome back to our recorded discussion. Still continuing about analytical techniques, instrumentation, and automation. Yeah, I know it's long because it's really long when it comes to your bishop. So although hopefully I was able to somehow summarize it in a way that you already know in which part to actually focus. Especially when it comes to analytical technique, instrumentation, and automation, we're actually going to cover three chapters. So we're actually halfway through the discussion, so hopefully I will also be able to help you understand the topics easier and better. Okay, so for this particular part, we're actually done with spectrophotometer, AAS, fluorometer, chemiluminescence, and electrophoresis. Now we're going to discuss your terbidimetry, your nephilometry, osmometry, electrochemistry, and then I think I would be doing a separate video for chromatography. And for the very last video, we'll be discussing about mass spectrophotometry and general principles with automation. Although no problem with automation because we've already started automation in your laboratory, so I was already able to discuss some of the key points or the driving forces that led to automation in the clinical laboratory. So with that, let's get started with our discussion so that we'll be able to finish up with these topics as soon as we can. So we're going to talk about terbidimetry and nephilometry. Please do remember, okay, so before I move forward, remember that when it comes to terbidimetry and nephilometry, I hope you still remember when we talk about your photometric spectroscopy. So there is a type of photometric spectroscopy that uses or that only measures the light, and there's also the other one, your spectrophotometry, okay? We have your photometry and we also have your spectrophotometry. So in your spectrophotometry, we dealt with light, spectrum, different wavelengths, and measuring a particular analyte. When it comes to terbidimetry and nephilometry, we're again going to measure your light, okay? We're again going to measure your light, but this time, we're measuring the light intensity or the light transmitted, but without considering your specific wavelength anymore, okay? So we're going to talk about terbidimetry and nephilometry, a very similar concept in clinical chemistry, but they are different from one another. And even though they are similar, there are key points for their differences, and I hope to highlight that in this discussion so that you'll be able to understand them immediately, okay? So let's get started. So first and foremost, always remember that both your nephilometry and terbidimetry are used to measure concentrations of larger particles. So large particles, what do we mean by that? This are actually for antigen and antibody complexes. So when we say antigen, antibody complexes, this is actually a bit more for immunology, although it's a good start because next semester we'll be having your immunology and serology around midterms for this specific topic. So hopefully you'll be able to still remember it by that time. So again, nephilometry and terbidimetry, we're measuring somehow larger particles. So we're measuring your antigen, antibody complexes, and generally your proteins, such as your pre-albumine or your transtyretine, and also several or other serum proteins. So as you can see, when we're talking about spectrophotometer, we are using that in the measurement of some of your analytes like glucose, your glucose, your enzymes, some of your lipids. And then for AES, we're measuring your elements, we're measuring a single atoms. When it comes to AES, similarly with your fluorometry and chemilaminescence, now when it comes to nephilometry, this is generally for larger particles. So I hope you get to compare the sizes of those molecules. So in here when we talk about your nephilometry and terbidimetry, again we're talking about measuring the concentration of large particles that because of their size cannot be measured through absorption spectroscopy. So meaning to say, your nephilometry and terbidimetry are for substances that cannot be measured through absorption spectroscopy. More specifically, what I want to highlight is that we cannot measure their absorbance. We cannot measure their absorbance. So that's what we mean by this statement. So because of that, if we are not measuring it through absorbance, how do we measure the concentration of these substances in the solution? So specifically for nephilometry, we measure the light, we detect or we measure the light that is scattered at far-used angles. So the scattered light yield a small signal and will be amplified in the measurement. So in the amplification part that is now your photo detector specifically, your photo multiplier too. So remember that in your nephilometry, the keywords here are the light scattered. Remember that in your nephilometry, we're not just measuring the light in a linear manner, but we're measuring it in several angles or in various angles. So remember that when your light hit a particular molecule, take for example an antigen antibody complex, instead of absorbing the light, it would scatter the light. So it would scatter the light in various angles. So in concept by principle, the more antigen antibody complexes are present in your solution, the more that the light will scatter. The more that the light will be scattered and the more light scattered, there is the more that you can measure the light and then that is nephilometry. In contrast, when we talk about turbidimetry, we're just simply measuring the reduction of light. So measures the reduction of light transmission due to particle formation, thus it detects a decrease in the large signal. So I would go again with my example last time. So if I have my flashlight, if I open my flashlight, the light being blocked there, the light being blocked or the reduction in the light is being measured in your turbidimetry. So simple. Okay, let me just show you a particular illustration so you would be able to understand it better. Here, I want you to look at, when it comes to your nephilometer, we're measuring light scattered by your particle. So we're measuring it typically within 90 to 15 to 90 degrees in angle. So as you can see from the light source, I want everybody to look at the screen. So you can see on the light source, you have your lens. So typically this would act like your monochromator. Okay, that would just simply concentrate the light into your cuvette. Your cuvette containing now your samples. Okay, containing now your sample. And as you can see, unlike the traditional one, if it is a linear, that would be for your turbidimetry. But I want you guys to focus on the detectors or the photo detector on the 90-degree angle and also on the 15-degree angle. So these two detectors, okay, detector 90-degree light scattering nephilometry. And you also have the detector of forward light scatter for your nephilometry. As you can see, we have multiple detector for your nephilometer. So that's one difference between nephilometry and turbidimetry. Okay, so it uses multiple detectors. Aside from that, what we again measure here are not the transmittance nor the absorbance, but rather we are measuring the light that is scattered. So as the light meet your antigen-antibody complex or your larger molecules like prealbumin and other protein, the light will now bounce off or the light will now scatter. And the light being scattered is the one measured by nephilometry, okay, or the one measured by nephilometer. So let me repeat myself with that again. The one that is being measured by your nephilometer or your nephilometry are your light scattered, okay, your light scatter. So as the light reaches a particular molecule or a particular complex, the light will scatter. And the light scattered at various angle, usually from 15 degrees to 90-degree angle, that is what we are measuring. In contrast to your turbidimetry, in turbidimetry, what do we measure? We measure the reduction of light transmission. Gaano kayo dami or gaano kakonte yung ilaw na hinaharang or tinatakpan nung ating complex, no antigen-antibody complex natin or no giban natin mga proteins. So in that sense, okay, if you're going to look at your illustration again from the light source, okay, you have the lens, you have the qubit and then after the qubit, you have now the detector, usually for spectrophotometer parayas and also your turbidimetry, ganan na ganan din pag ating sa turbidimetry, okay. So ganan din pag ating sa turbidimetry. The orientation of the components are similar to your spectro. Linear lang talaga sila. So again, light transmitted in the forward direction is the one being detected. So we're not measuring the other light that are scattered in any angle. We're just simply measuring the one at 180 degrees, okay, yung straight lang, yung linear lang. So in here, the amount of light absorbed by the suspension of particle depends on your specimen concentration and also your particle size. So unlike in spectrophotometry, okay, the amount of the absorption, the absorbance is represented by your concentrate is directly proportional to your concentration. In here, in turbidimetry, not only is your concentration the one affecting your absorbance but also your particle size. So mas malaki yung particle, mas madami yung na absorb niya na light, okay. That's why hindi ito tulad ng spectrophotometer in natin, okay, hindi ito tulad ng spectrophotometer in natin. Because again, if you guys could remember beer slow, the molar absorptivity is constant. In turbidimetry, hindi yung constant kaya hindi beer slow, pagdating kay turbidimetry ha. So again, when we're talking about turbidimetry, if this is the light source, okay, let me just open na lang my flashlight. So if this is the light source, okay, if this is the light source and this is your qubit, okay, take for example this one's your qubit. So the light that is being blocked, okay, kung gaano ka dami, okay, kung gaano ka dami yung nag-pass-through, okay, kung gaano ka dami yung nag-pass-through, yung yung minime-measure natin sa turbidimetry. Okay, yung yung minime-measure natin sa turbidimetry. So again, makikita na tayo ha, turbidimetry, it measures the reduction of light. In your nephilometry, it measure the light scatter, okay, the light that is scattered at values ang get. So if I'm going to go back to that particular slide, I want you guys to remember this because this will already take you far when it comes to nephilometry and turbidimetry. So let's just try to differentiate them further. Again, when we're talking about nephilometry, we're measuring light scatter, okay. Again, nephilometry, light scatter. In turbidimetry, we're measuring the reduction of light, okay. Second difference between the two. In nephilometry, your photodetector, specifically your photomultiplier tube are situated in various angle within 15 degrees to 90 degrees angle. When it comes to your turbidimetry, it is in linear form. So you have your light source, your monochromator, your qubit, and you already have your photodetector also in the form of your photomultiplier. So always remember that, okay. Similarity, your nephilometry and turbidimetry are usually being used when we're talking about larger molecules because when it comes to this larger molecules, not only is the concentration affecting the light being scattered or the light being reduced but also the size of your particle, okay. We're also now taking into consideration the size of your particle. Kasi mas malaki yung particle, mas madami yung inaabsorb niya. And that would contradict now the molar absorptivity when it comes to beer flow. That's why we cannot use your spectrophotometer. Okay, we cannot use your spectrophotometer. Okay, so moving forward. So those are actually your nephilometry and your turbidimetry. So with that, okay, I hope I made myself clear when it comes to turbidimetry and nephilometry. So if you have any questions or clarification, please feel free to send me a message. So that is it for nephilometry and turbidimetry. I think this video into segments so that you'll be able to only go back to turbidimetry and nephilometry if you would want to. But I will also be discussing your osmo metri right after this. So if you have any questions, so please send me your messages or send me your message regarding your question. So that is nephilometry and turbidimetry. Thank you so much for listening.