Subtitles section Play video Print subtitles >> Sticks. All right, but we're going to talk about spins today. Not about sticks. So I want to continue our discussion of the concepts and theory behind NMR spectroscopy and again, this is not going to be about math or anything to that extent. But we're going to be thinking very, very qualitatively. All right, so when we last left things we had said that there are two spin states for a dipolar nucleus like a proton or C13. There's spin up and spin down, and if you have an applied magnetic field there's a small energy difference between the spin up state, what we're calling the Alpha state and the spin down state, what we're calling the Beta state. And because that energy difference is so small unlike IR spectroscopy or electronic spectroscopy, UV Vis spectroscopy where the energy differences are very big and all of your molecules are in the ground state, here there's only a miniscule number of nuclei in the lower state more than the number in the upper state. We said if there are -- if we take 2 million protons, out of those 2 million protons depending on the applied magnetic field, it will be 50 or 80 or thereabouts difference in population. In turns out that difference in population is going to be extremely important because it is only that differential population that's going to be able to get us a signal. All right, so if we think about things in an XYZ coordinate frame, and I'll talk more about NMR spectrometer in a second and how it works. But imagine for a moment we have some coordinates, so the X coordinate is coming out of the plane, the Y coordinate is in the plane and the Z coordinate is pointing up and we're going to have our applied magnetic field BNOT pointing upwards. That kind of makes sense. These super conducting magnets are always vertical because you've got this big pot of liquid helium surrounded by a vacuum vessel, surrounded by liquid nitrogen, surrounding by a vacuum vessel. And, those small amounts of population, that small differential of population with spin up is going to give rise to a net magnetization. Now, in other words a way to think about this is for most of our cases we're going to have one nucleus pointing up, one nucleus pointing down in their spin and there's no net vector here. Those vectors cancel each other out. But for that small differential of access vectors you're going to have some net magnetization along the Z-axis. Now, the ay it works, when you apply a magnetic field is you actually have those vectors that are processing around. So in other words they are processing at that resonant frequency, at the Lamar Frequency at 500 megahertz for 117,500 Gauss magnet. So we actually can represent this by saying okay, we've got spin sort of pointing in every which way, and I'll just draw two directions. They're all processing around. So remember, remember this is only the differential population that we're worried about because already for every one where you have one up and you have an opposing one spin down those vectors are going to cancel each other out. Now the other thing is they're not quite on axis. In other words they're not like this. It's like a gyroscope if you've ever hung it from a string, the gyroscope doesn't -- who's hung a gyroscope from a string as a kid? The gyro in physics lab or something. The gyroscope doesn't hang vertically, it kind of hangs off axis and goes around like this. But if you think about it, since those spins are not bunched up for every one that's processing like this there's another one that's opposite it. So in other words if were just like this you'd say, oh there's a net magnetization along the Z axis but also a net magnetization along the Y axis. But there are other spins that are like this and they're all going around. So everything is canceling out except the net magnetization along the Z axis. All right, what I want you to imagine right now is that we're going to place a coil along the X axis and we're going to put energy into that coil. We're going to apply a magnetic force. And I want you to think classically because the quantum mechanical thing is going to be we'll flip the spins. I'll show you that in a second. But you have your net magnetization along the Z axis, and think back to classical physics. If I apply a force along the X axis, right hand rule and all of that good stuff, we rotate our vector downward. So after we apply a pulse, I'll just say a pulse. If here's out net magnetization, when we apply an RF pulse our net magnetization moves along the Y axis, and so I guess if I want to actually represent it I'll just say XYZ, and I'll say here's our net magnetization. And as you'll see in a moment we're going to have continued procession. And again, if you're worried about the fact that all of our vectors are not lining up, that they're all processing like this, just think as I apply a pulse and drive my magnetization from the Z axis onto the Y axis, the vector sum is right along the Y axis even though there are some that are like this, I drive it down. They're countering each other. There are some like this, I drive it down. They're countering each other. And so our net magnetization ends up a long the Y axis. Does that make sense? All right, lets come back to our spins to see what this means. So, the way I was trying last time to represent this very small difference between the Alpha state and the Beta state was to show some vectors, some spins pointing up in the Alpha state. And some spins pointing down in the Beta state, and to try to represent this miniscule difference in population what I did for the purpose of my drawing was I drew 6 with spin up in the alpha state and 4 with spin down in the Beta state. >> [Inaudible] nuclei right? >> Those are representing exactly the spins of individual nuclei. So in other words, if we had a mole of -- or more realistically if we had a millimole of CHCL3, proiochloroform [assumed spelling] in our NMR tube, what this would represent would be the different, the nuclei of the hydrogen there and we would have out of that millimole of nuclei we would have a small access in the Alpha state and they would all be processing. All right, so if we apply an RF pulse, and now I'm going to be a little bit specific, if we apply a pulse long enough that it's what's called a 90 degree RF pulse or a pi over 2. That's just radians and degrees, your choice and they get used interchangeably. What that does is it equalizes the population of Alpha and Beta state, so I'll represent that by 5 spin up and 5 spin down. And this situation is exactly the situation that we have at the end of my little drawing over on the left-hand blackboard. In other words here's our net magnetization. And so the key is now we have no net magnetization spin up, no net magnetization spin down, but the very important point is we have the net magnetization focused along the Y axis. It is not diffuse, it is not pointing in all directions. We actually have net magnetization in the XY plane. And if we apply a longer pulse, a more powerful RF pulse, so again I will represent our 6 little arrows and 4 little arrows representing our differential populations of the Beta state. If we apply a more powerful RF pulse, what we call a pi RF pulse or a 180 degree RF pulse I can invert the population. In other words I will represent that by 4 arrows pointing up and 6 arrows pointing down. And if I want to draw that on my diagram, can anyone tell me what I do with my net magnetization on my little XYZ diagram at this point? >> Down. >> It's going to point down, exactly. All right, and this is the damming thing. Well, one of the many damming things about NMR spectroscopy is no matter what you do with your pulses you are limited to the difference in population that occurs between the Alpha and Beta state, and later when we start to talk