00:00:00 This is a picture of the laboratory at Jones Chemistry Building where the early microchemistry

00:00:11 work was done on plutonium during the war. We worked with microgram amounts and here's

00:00:18 another instrument, a quartz fiber microbalance, just a quartz fiber here with another quartz

00:00:29 fiber coming down here to the weighing pan and you put the sample on that and the amount

00:00:35 of depression of the quartz fiber related to the weight and with this Burris Cunningham

00:00:40 who was the first man to isolate plutonium in the Metallurgical Laboratory during World

00:00:45 War II and the first person to weigh it which occurred on September 10, 1942, 2.7 micrograms

00:00:56 was performed with this balance. This is another example of the kind of crude apparatus we

00:01:05 used. In the discovery of the two elements with the atomic numbers 95 and 96, in certain

00:01:13 bombardments we produced alpha particle emitters and we had a counter up here and then put

00:01:23 the sample here and then placed mica sheets in between to measure the energy of the alpha

00:01:30 particles. The more energetic the alpha particle being, the more mica sheets you had to interpose

00:01:39 and then plotted the range of these particles and we found a long range particle like this

00:01:46 in the summer of 1944, July 15th, 16th.

00:01:51 It's probably more important than what I was saying.

00:01:59 This represents then the discovery of element 96 which we named Curium. That's just how

00:02:10 simple the apparatus was then, just measuring the range of the alpha particles with thin

00:02:16 sheets of mica and then a little later in January, and this is a reconstructed diagram

00:02:22 but it's just exactly how I presented it at a meeting at the Metallurgical Laboratory

00:02:27 on January 31st, 1945, showing the absorber thickness of mica and then how there was

00:02:35 a long range alpha at 4.0 centimeters of area equivalent to it, that corresponded to element

00:02:42 95 Americium and 4.8 centimeters area equivalent that corresponded to the element Curium.

00:02:50 This is the group that I brought with me from my class at UCLA to work with me at the Metallurgical

00:02:59 Laboratory. I had to find chemists anywhere I could find them. None of these knew anything

00:03:05 about nuclear chemistry but I convinced them to come to Chicago and we quickly taught them.

00:03:12 This is just an example of how we were required to keep our notebooks for patent purposes,

00:03:26 our disclosures and the witnesses. This is something I wrote on October 30th, 1942. This

00:03:33 is an example of Burris Cunningham's laboratory showing where, how it was necessary to keep

00:03:42 in these so-called dry boxes due to the necessity for protection against the radioactivity.

00:03:49 Now I'm quickly running through this and then I'll be finished. This is a picture of the

00:03:55 first group to attend the Gordon Research Conference on Nuclear Chemistry, which was

00:04:00 held in New Hampshire in 1952. If we had more time I could identify for you on this picture

00:04:08 all of the prominent nuclear chemists in the United States. This is another picture at

00:04:15 the Gordon Conference in 1954, 1956, 1957 and then two more pictures just to show you

00:04:27 more modern apparatus that we use in nuclear chemistry. This is from my laboratory in Berkeley.

00:04:35 This is just a picture taken from the corner of the room where we make our measurements

00:04:40 on the decay of radioactive isotopes by alpha particles and spontaneous fission. And this

00:04:46 is a picture of the corner, another part of the room where we make our measurements with

00:04:53 more modern apparatus on electrons, that is beta particles and gamma rays. Thank you very

00:05:02 much.

00:05:03 We have very little time, but it's one question or two brief ones if you have any.

00:05:23 It's a great occasion to be with you.

00:05:27 Well, all right, thank you very much. I'm going to be frightening people by talking

00:05:32 about the shortage of time. Well, thank you very much, Glenn, and we now proceed to the

00:05:39 next paper, which is by Professor Mildred Cohn. Professor Mildred Cohn is a professor

00:05:48 emeritus of the University of Pennsylvania. She was trained in Harold Urey's laboratory

00:05:57 or took her PhD there in Columbia University in, I guess, physical chemistry, but moved

00:06:04 pretty quickly into this field, which has been her field ever since. And her field has

00:06:12 really become a biological chemistry, and she's pretty much stayed with it. She's received

00:06:18 many honors and many awards. One of many of them is the United States National Medal of

00:06:24 Science. Her topic for us today is from do-it-yourself to high-tech in biochemistry. Mildred Cohn.

00:06:33 This month, actually, is 50 years since I went into the field of biochemistry. It's

00:06:47 also 50 years since I defended my thesis at Columbia University, and I never stayed with

00:06:53 physical chemistry. I went immediately into biochemistry. One of the reasons for this

00:06:58 is that the laboratory I was working in, Professor Urey was enriching isotopes, stable

00:07:04 isotopes. He, of course, had already gotten the Nobel Prize the year before I started

00:07:10 working with him for discovering deuterium, but he was in the process of enriching C-13,

00:07:17 O-18, lithium-7, and so on. And at that time, the biochemists, very few of them, I will

00:07:25 admit, were becoming aware of the fact that one could do really wonderful things with

00:07:30 isotopic tracers. And this field was being pioneered by Rittenberg and Schoenheimer.

00:07:38 Rittenberg was a former student of Urey's who preceded me by about two years. I still

00:07:45 remember the example that Schoenheimer used to give in his lectures back in 1938 or so

00:07:53 on what made isotopes so wonderful as tracers for metabolic processes in animals. He said

00:08:05 that before one had isotopic tracers, one would feed a compound to a rat and then look

00:08:13 at the urine and decide what was in it. And he said that was the same as putting a nickel

00:08:18 into a vending machine and getting a piece of chocolate. But, of course, with the tracers,

00:08:23 you didn't make such errors as to say that the nickel had been converted into chocolate.

00:08:28 But he, Vincent de Vigneault was very interested, he was an outstanding biochemist who was

00:08:39 interested in amino acid metabolism, and he decided this was the time for him to go into

00:08:44 the field of isotopic tracers. And since there were very few people who were trained

00:08:51 at that time who knew anything about how to measure stable isotopes or how to handle them,

00:08:57 I was fortunate to get the position. But I do want you to get a notion of what it was

00:09:03 like to be a graduate student in physical chemistry in the middle 30s. It was a strictly

00:09:10 do-it-yourself kind of thing. And we just assumed that we did anything that was required

00:09:16 of us. For example, I had to set up a method of measuring D2O and H2O18 by their density.

00:09:28 This required building a constant temperature bath that was good to a thousandth of a degree.

00:09:32 Well, I just went to the literature and found out how to do it and did it. I think the extreme

00:09:37 is a fellow graduate student of mine, T. Ivan Taylor. His thesis problem was to separate

00:09:42 the isotopes of lithium-6 and lithium-7. He did this by building a 35-foot column, a steel

00:09:50 column, which he filled with zeolite and took advantage of the fact that there was the equilibrium

00:09:59 between lithium chloride, actually in the end he used lithium bromide, that was even

00:10:04 better, the partitioning between the solution and the solid was favorable for separation.

00:10:12 And then at first he had his samples measured by a mass spectrometer that they had down

00:10:20 at the Nitrogen Fixation Laboratory, Brewer's Laboratory in Washington. But after a while

00:10:27 that got sort of burdensome, so he went down there and spent a week and he came back and

00:10:32 he built a mass spectrometer. And he was just a graduate student and nobody thought

00:10:36 that was at all unusual. Well, then when I went into biochemistry, the biochemists had

00:10:44 the notion that if you were a physical chemist you could do anything. For example, they asked

00:10:49 me to modify the telephone circuit that they had. And then they asked me, oh they had a

00:11:00 broken leads and northern suspension galvanometer, they wanted me to fix that too. Well, in any

00:11:05 case, I did set up a method for measuring D2O, the same one I had used before, the falling

00:11:12 drop method. And a few years later it was decided that what we really needed was a mass

00:11:19 spectrometer. By this time, World War II was on, and this was about 1942 or so, and the

00:11:26 problem came up, there were no instruments available commercially, I should add. So I

00:11:33 had to build one, and I did, with the help of Harry Thode in Canada, who had designed

00:11:38 a new mass spectrometer and was kind enough to make the inside works for me, and I just

00:11:45 put it all together. Now, I do remember the kind of problems you had. For example, I had

00:11:53 to spot weld platinum wire to the nichrome plates which were in the ionizing chamber

00:11:58 of the mass spectrometer, and I found out that to buy a timer, which I could use with

00:12:04 this tiny spot welder which I had made from a test tube holder, would cost $1,200. I only

00:12:13 had four spot welds to make, and of course it was out of question spending $1,200, that

00:12:17 was a fortune in those days. So I went to a friend of mine who was an electrical engineer,

00:12:22 and he said, are you willing to spend a nickel on each spot weld? And I said, sure. He said,

00:12:26 well, all you have to do is buy a fuse, and they're wonderful, he said, they are absolutely

00:12:30 reproducible, they will blow depending on whether it's 5 amps or 10 amps in a given

00:12:37 time. I did have a transformer that gave me enough current, you see. So that was the kind

00:12:41 of sort of do-it-yourself thing we had to do in those days. It was World War II that

00:12:47 made all the difference in the availability of instrumentation. That's when various circuits

00:12:57 for measuring radioactive isotopes, I should say, in the years during World War II, we

00:13:03 had to build our own Geiger counters if we wanted to use any radioactivity. It was only

00:13:09 after the war that instrumentation that had been developed during the war became available,

00:13:15 and that's when work with radioactive traces took off. But there was still no mass spectrometer

00:13:22 on the market that could be used for measuring isotope ratios. Al Nier had developed some

00:13:29 very beautiful ones during the war, and in 1946 I moved to Washington University and

00:13:34 set one up there with his help, again, for the essential ionizing chamber and the whole

00:13:40 tube, and that fortunately was the last mass spectrometer I had to build. The next one

00:13:46 I got, I bought. But that one worked for, I used it for 14 years, the 14 years I was

00:13:53 at Washington University. Now, I really didn't want to talk that much about isotopes, except

00:14:00 that my early experience in do-it-yourself was with isotopes mainly, but I really want

00:14:07 to talk about the field that I've been working in in the last 15, 20 years, and that is

00:14:14 nuclear magnetic resonance. That has a very interesting history because of its use in

00:14:21 biochemistry and also in medicine, and I would like to talk a little bit about that.

00:14:28 A Beckman DU spectrometer was treated as something very special. When I got to Washington

00:14:34 University in 1946, there was one in the whole department, and you had to sign up to use

00:14:40 it. By 1953, there was one in every laboratory in the department, so things moved pretty

00:14:46 fast as far as instrumentation, but at that time, this was indeed the most useful region

00:14:53 of the electromagnetic spectrum for studying biochemical compounds and so on. Now, in

00:15:00 1946, it was a better year, too, because that was the year that Lockett Stanford and Purcell

00:15:06 at Harvard discovered nuclear magnetic resonance. Actually, it had been observed first by Ravi

00:15:13 in 1938, but that was in a molecular beam. In bulk matter, it was first discovered in

00:15:21 1946. Now, as you can see from this, actually, this is quite out of date because of the frequency

00:15:30 that one can get to now is about an order of magnitude higher than this, up to 10 to

00:15:36 the 8th, where the commercial instruments are now at 600 megahertz, so this was made

00:15:42 in the early days when one had 30 and 40 megahertz instruments. You see where it is

00:15:51 on the energy scale. The transitions between two levels in the nuclear magnetic resonance

00:15:59 has very, very small energy difference. This is both good and bad. It's very sensitive

00:16:06 in the sense that very small changes in the environment of the nucleus are observable.

00:16:14 On the other hand, you need an awful lot of nuclei to see a transition because the energy

00:16:22 is so small and there's such a small difference in the population of the two energy states,

00:16:27 so that we have this marvelous region of the electromagnetic spectrum, which is very sensitive

00:16:35 to the chemical environment of the nucleus, but from the biochemical point of view, one

00:16:39 of its great disadvantages is that you do need a lot of material to do it. Now, particularly

00:16:45 in biochemical compounds, particularly macromolecules, there is very little energy difference between,

00:16:53 let us say, an enzyme that is active and an enzyme that is inactive, as far as the whole

00:16:59 molecule is concerned. It's the fact that it takes so little energy to go from one state

00:17:05 which is active to another which is inactive. Those are the kinds of things that can be

00:17:10 observed in nuclear magnetic resonance.

00:17:13 Now, in the next slide, I have the development of instrumentation and what it all meant.

00:17:28 Now, in nuclear magnetic resonance, of course, the frequency, the difference between the

00:17:34 two lattices is, of course, proportional to the magnetic field. The sensitivity, the intensity,

00:17:42 is proportional to the square of the magnetic field in which you place the material, so

00:17:49 that the higher the magnetic field, the more sensitive the measurement and, of course,

00:17:58 the resolution increases because the chemical shift, which is due to the difference in electronic

00:18:05 environment of the nucleus, is proportional to the field, so that we get both increased

00:18:12 resolution and increased sensitivity as you go up in frequency. That means going up in

00:18:18 magnetic field.

00:18:19 Now, the first experiments that were done by the physicists Bloch and Purcell were both

00:18:25 done with permanent magnets. Purcell had a field of about 7 kilogals and Bloch had a

00:18:35 field of about 1.8 kilogals. But, of course, they weren't interested in resolution. In

00:18:41 fact, in 1951, Arnold and his co-workers in Bloch's laboratory were the first to show

00:18:52 that you could resolve three peaks from ethyl alcohol, the CH3 group, the CH2 and the OH,

00:18:58 and that was the beginning of interest in chemists in nuclear magnetic resonance.

00:19:05 I should also add that it was the end of Bloch's interest. He said it was no longer physics,

00:19:09 it was now chemistry.

00:19:10 And, of course, because of the need for more resolution and not so much sensitivity, chemists

00:19:24 fortunately usually have plenty of material, but biochemists don't. Biochemists are the

00:19:28 ones who are always fighting sensitivity problems. But both are interested in increased resolution.

00:19:35 And as the years went on, the first commercial instrument became available from Varian in

00:19:41 about 1955, and it was rather low. It was down around 40 megahertz. If one looks at

00:19:51 the first book that was written on nuclear magnetic resonance for chemists by Bernstein,

00:19:59 Schneider and Popel, all the spectra that they show were taken either at 30 megahertz

00:20:04 or 40 megahertz, because that's all that was available at the time.

00:20:08 Varian then put out, in about 1957, they put out an instrument that was 60 megahertz, and

00:20:15 that was a workhorse for many, many years until it was increased to 100 megahertz in

00:20:25 about 1961 or so. And that is the limit to which you can make a magnet. By that time,

00:20:34 you see, you're at about 23 kilogals, and that's the limit of an iron core magnet.

00:20:41 And so there it would stay, unless one could increase the magnetic field in some other

00:20:48 way. And that's when superconducting magnets, oh, I think it was 1967 that Varian introduced

00:20:59 a superconducting magnet. The proton-free magnetic resonance frequency for that is 220

00:21:06 megahertz. That was the first one that was made. I should mention there was another company

00:21:12 besides Varian that made instruments back here in about the late 50s, and that was Truver

00:21:21 and Taub in Switzerland. But that company never seemed to catch on. Their magnet was

00:21:28 much smaller. You could only do 30 megahertz protons on it. And it eventually was bought

00:21:37 by Bruka, which, as you know, is now one of the big companies that makes commercial

00:21:42 instruments. When you got to 220 megahertz, then it was possible to get sufficient resolution

00:21:51 and sufficient sensitivity to do macromolecules such as ribonucleus. I'll show you some spectra

00:21:57 in a few minutes. And I should tell you that since that time, it's been a steadily increasing

00:22:05 rate of growth. You will notice that this scale is logarithmic. And now the commercial

00:22:11 instruments go up to 600 megahertz, which is about 142 kilogauss. I should point out

00:22:18 that for a long time, the physicists had superconducting magnets. But here the requirement for homogeneity

00:22:24 of field is what's so important. You need at least one part in 10 to the eighth and

00:22:30 preferably one part in 10 to the ninth homogeneity. And that is not easy to come by. And that

00:22:36 is one of the things that was necessary to work on the design of magnets that could do

00:22:44 that. And when you get up to 600 megahertz, you really have to worry about the kind of

00:22:50 materials that you use in winding such a magnet. In this range, electromagnets are used. And

00:22:58 permanent magnets really cut off much lower than even the electromagnets. But nowadays,

00:23:04 practically no one uses anything but the superconducting magnets. And I'll show you the difference

00:23:11 in what you can see. The first protein that was ever looked at is ribonucleus. Ribonucleus

00:23:17 has 124 amino acids. It's got 684 hydrogens that are carbon bound. And all they saw, this

00:23:31 was the first one, was done by Saunders, Wisnia, and Kirkwood with a 40 megahertz instrument

00:23:39 and all they got were these four groups of peaks. And it was shown by Dardesky who did the amino acids

00:23:47 that this consists of, that this could be explained more or less by the addition of the spectra

00:23:53 of the constituent amino acids. Now by 1980, two developments had taken place. The field

00:24:02 was up so that the proton frequency was now 360, it was a superconducting magnet. And

00:24:08 another very important thing, Fourier transform spectroscopy had been introduced. In the old

00:24:16 instruments, one had to go sweep through the magnetic field, one kept the frequency constant

00:24:23 and swept through the magnetic field in order to get each different proton in resonance.

00:24:29 And that meant that you looked at one peak at a time. Now with Fourier transform, the

00:24:37 great thing about it is that in one pulse, you excite the whole spectrum so that you

00:24:43 spend no more time looking at all the protons than you did originally at one proton. Now

00:24:52 if your compound has only one proton, there's no great advantage to Fourier transform. But

00:24:57 if it has 684 protons, it certainly is an advantage. Another development that had taken

00:25:03 place that was very important in making high resolution NMR spectroscopy useful to biochemists

00:25:11 was the fact that computers and computer programs had been developed. The 360 megahertz

00:25:19 instrument comes with a dedicated computer as do all the others that are higher or lower

00:25:24 than that that are superconducting magnets today. And by manipulating the data, you can

00:25:30 get resolution enhancement and this spectrum goes over to that one. And this is only, as

00:25:36 I say, I happen to have one at 360 megahertz, the highest that is readily available today.

00:25:43 Many laboratories have 500 megahertz and you would get even better resolution than you

00:25:48 do. Another thing that's been introduced, and this again is a matter of developing pulse

00:25:53 frequencies and having excellent computer programs and a big computer to do all the

00:26:00 calculations for you, is 2D NMR where it's done in two dimensions. But I haven't got

00:26:05 time to go into that. From that one gets actual distance if you use it in connection with

00:26:11 various and sundry things like nuclear overhauser effects and study the coupling constants and

00:26:16 so on. One can get distances, let's say, from the amide proton on a particular amino

00:26:24 acid to the alpha, the hydrogen on the alpha carbon. And from that one learns something

00:26:31 about the structure of the protein. So far it's been limited to molecules of molecular

00:26:41 magnitude under 10,000, but people are now working up toward 15,000 and getting a complete

00:26:47 story on that. Now, another element that is very useful to do NMR on is phosphorus. If

00:26:56 one is interested in studying metabolism, phosphorus-31 is a good, it's not of course

00:27:02 as sensitive as protons. It has only 6.6% the sensitivity of protons at the same magnetic

00:27:09 field. But on the other hand, it's 100% of one isotope phosphorus-39. In continuous

00:27:18 waves NMR in 1959, I had to use 500 millimolar ATP in order to get a high resolution spectrum.

00:27:27 And I should tell you this, using Fourier transform in 1976 with three times the field,

00:27:34 this was a 180 megahertz instrument, this was a 60, this is what I got with one millimolar.

00:27:41 Now, the reasons for the difference of a factor of 500 between these two is, first of all,

00:27:48 this was done only in 5 millimeter tubes. In order to get high resolution, one spins

00:27:53 the sample, and Varian had not developed a spinner for anything bigger than a 5 millimeter

00:27:59 tube. At this point, we were using 10 millimeter tubes, so of course we had much more volume,

00:28:04 many more nuclei in the sample, and that helped. In addition, we were using Fourier transform,

00:28:10 and at this time, in 1959, you could only take one scan. By 1963, they did introduce

00:28:18 a computer of average transients. By this time, in 1976, of course, there was no problem

00:28:24 with optical scans, and so one could easily do 1 millimolar ATP, as easily as one could

00:28:31 do 500 millimolar in 1959. Now, most of these instruments have been,

00:28:43 most of the work has been done with commercial instruments. However, the developments often

00:28:48 take place either in academic laboratories or in other places that are not instrument

00:28:55 makers, but some of them, of course, do take place in the companies that are instrument

00:29:00 makers. But there is a set of mind, or was, of scientists in 1959 that is really interesting.

00:29:11 I remember I sent a paper into JACS. It was a communication, because I got all excited

00:29:17 when I got this and found the effect of both diamagnetic and paramagnetic ions on this

00:29:23 spectrum. That's actually what I was interested in. I was interested in the effect of metal

00:29:27 ions, because all the reactions that ATP, which is adenosine triphosphate, is the central

00:29:32 compound of bioenergetics in the cell, and all the reactions of which it's a substrate

00:29:41 requires a divalent metal ion. And being a physical chemist at heart, I chose the simplest

00:29:46 thing. What does the divalent metal ion do? In any case, I sent in a communication to

00:29:51 the Journal of the American Chemical Society, and it was rejected. And one referee said

00:29:56 that the chemists wouldn't be interested in this, because I had used a commercial instrument.

00:30:03 The second referee said that no biochemist would understand it and that I had better

00:30:06 expand it into a full paper, which I did. But in any case, that was a little disturbing

00:30:14 that they thought that if you did it on a commercial instrument, the chemists weren't

00:30:19 interested. But of course, this was early days when most of the chemists who were working

00:30:23 in nuclear magnetic resonance were indeed using their own homemade things.

00:30:28 Now, the highest frequency that I have ever done phosphorus at is in the 600 megahertz

00:30:38 instrument that was developed at Carnegie Mellon by Botherby and Dada. And they asked

00:30:44 me, was there anything I could give them that would show off their instrument? And so I

00:30:49 gave them some O-18 ATP labeled with O-18. And the chemical shift due to the isotope

00:30:56 effect of O-18 on the phosphorus is of the order of 0.02 parts per million. And as you

00:31:03 can see, this is the triplet. And it's 63% O-18, so one has many species. One has phosphates

00:31:11 with one oxygen, two, three, four, and zero oxygen, 18. And this is a spectrum of that.

00:31:20 And every one of them is resolved. One even resolves the difference between a bridge oxygen

00:31:30 and a non-bridge oxygen. But that is done on that instrument. And their instrument was

00:31:38 not really terribly useful in a way because it doesn't have a persistent field. The field

00:31:45 is only temporary. Now they have developed 600 megahertz instruments commercially. Several

00:31:52 companies have them that are persistent. Now, this comes from a paper by Pines in the

00:32:02 Council of Chemical Research showing the year and the cost of these various instruments.

00:32:10 And you'll notice it's an exponential rise. And by this time, 500 is in many, many laboratories.

00:32:21 And you see, it costs about $600,000, depends on how many accessories you want. The 600

00:32:28 is now available, and I don't know just what the price is, but it's probably somewhere

00:32:33 around $900,000. Now, this is the kind of thing that I think one should give some thought

00:32:40 to about how it was when you did it yourself and now when you have to use high technology.

00:32:46 There are certainly advantages to the high technology. You acquire data in a week that

00:32:51 you would think would take you six months. But on the other hand, you do have to be financed.

00:32:58 I mean, one can't buy this kind of instrument in an ordinary laboratory. And of course,

00:33:04 back when I started in 1937 in biochemistry, there was no NIH, there was no NSF. The government

00:33:13 was not supporting research. And people started in a laboratory with maybe a few thousand

00:33:20 dollars. That's out of question today if you want to do this kind of work. And so where

00:33:27 it's all going to end, I don't know. At the moment, there's a 900 megahertz magnet being

00:33:34 designed by Dadock at Carnegie Mellon and another one on the West Coast. They're not

00:33:41 yet operational, but I'm sure they will be, and they will probably be well over a million

00:33:48 dollars. And then, of course, one has to have the liquid nitrogen and the liquid helium

00:33:52 with these being superconducting magnets. And so far, I've talked only about high resolution

00:33:57 work in biochemistry, which is very good for macromolecules and also for small molecules

00:34:06 that are of biological interest. But there have been developments in the last seven years

00:34:11 or so where it's actually being used in intact cells, tissues, and intact animals. There

00:34:20 are two kinds of things that are being done. One is spectroscopy, the kind of spectroscopy

00:34:25 I've been talking about. Phosphorus-31 is very useful because there are many phosphorus

00:34:31 compounds that tell you about the bioenergetics and the metabolic state of the object you're

00:34:39 looking at, if it's an organ or a cell. And also, there's no interference from lots of

00:34:47 other things. Protons are pretty hard to do because there's too much information. I mean,

00:34:51 it's just overwhelming the number of lines you get. And then, of course, there's the

00:34:55 water peak, which is so tremendous in any tissue. Of course, there are ways of getting

00:35:01 around that, and they are being worked on. But so far, the most popular one has been

00:35:07 phosphorus to look at in an intact cell or tissue or animal. And I just want to show

00:35:14 you, this was done with a single barnacle muscle cell. Now, I will admit that this is

00:35:21 a very unusual cell. A barnacle muscle cell is one millimeter in diameter, and it's about

00:35:27 oh, an inch and a quarter long. Now, that's the most unusual cell. But with one single

00:35:33 cell, you can look at the phosphorus NMR. This is the ATP again, the beta of the ATP,

00:35:42 the alpha and the gamma. This is phospho-arginine, and then there's a little bit of inorganic

00:35:49 phosphate here. I should point out that the first work of this kind that was done was

00:35:53 done by Moon and Richards, again from Caltech, who looked at red blood cells, and they pointed

00:36:01 out that the inorganic phosphate, which they saw in that, was a good measure of pH. So

00:36:06 one can measure the pH of an intact cell by the, since the chemical shift of inorganic

00:36:15 phosphate is a function of pH, one can tell from the chemical shift what the P in muscle

00:36:23 and in other tissues as well. In invertebrates, such as barnacles, it's phospho-arginine instead

00:36:29 of phospho-creatine. And this was done, actually, on the ordinary high-resolution instrument

00:36:36 by putting the cell inside a little tube and winding the coil, the receiving coil, around

00:36:43 it, and it was superfused during the experiment, and that's how the experiment was done in

00:36:50 a, actually in a 360 megahertz instrument. However, if you want to look at a human being,

00:36:57 they are now whole body magnets, large enough for, okay, large enough for to put in a whole

00:37:05 human being, and you can look at the arm of an individual or his brain, and as you can

00:37:13 see, you get characteristic patterns. And here you even see the triplet, the beta triplet

00:37:19 resolved almost, and this is the gamma phosphate, which is doublet, which is resolved. And lastly,

00:37:25 I'd like to mention imaging, which is, of course, now called MRI. The physicians changed

00:37:33 it from NMR to MR because it seems the public didn't like the word nuclear. So, that all

00:37:40 started, there was a man by the name of Gavillard who pointed out that the information you got

00:37:44 could encode space as well as frequency, and the first one, this is Lauterburg, however,

00:37:53 in 1970, that was in 1952. In 1973, Lauterburg did an experiment with two tubes, one millimeter

00:38:00 in diameter each, and showed that he had a field gradient. Obviously, the one over here

00:38:06 would resonate at a different frequency than this because the fields are different, and

00:38:11 he used those to show that it would work, and indeed it did, and here is an example

00:38:16 of a human brain image that was done here at the hospital in the radiology department

00:38:25 at the University of Pennsylvania Hospital. And this is a normal brain, and this is a

00:38:31 brain with a tumor in it, which you can see right here. And I will stop here and tell

00:38:37 you that now almost every hospital, any large hospital in the country, certainly has at

00:38:43 least one such instrument, and it is improving all the time, and in the future what they

00:38:49 will have is one instrument that can do both spectroscopy and imaging, and that is already

00:38:55 being done and will, I'm sure, continue to be perfected. The amount of technological

00:39:01 improvement in these images has been fantastic in the last five years, and that's due to

00:39:06 the engineering efforts of the various companies that put out these instruments. Thank you.

00:39:11 Well, thank you very much indeed. It's really important to see what has happened to biology

00:39:29 from this instrumentation, and that's a particularly lucid and interesting discussion. I'm told

00:39:38 that another part of video discussion with Arnold Beckman is available and will be shown

00:39:48 as a break before the break, so that if the TV people would show this brief thing and

00:39:56 maybe ask Arnold Beckman for a comment or two.

00:40:00 Well, Jack, I think it's only fair to warn you right from the start here now that you're

00:40:03 talking to a fellow who certainly admires but just doesn't begin to comprehend the complexities

00:40:07 of this great field with which you people here at Beckman are so familiar.

00:40:11 Well, Ken, I don't think it's really so complicated. I think we can give you a pretty clear picture

00:40:15 here.

00:40:16 You really do?

00:40:17 Sure, I think so.

00:40:18 Well, all right, supposing you try to convey to me just what this flame photometer does.

00:40:22 Well, the flame photometer is used by doctors primarily. It's used in the medical field,

00:40:27 and one example is taking blood samples down here where the blood is sucked up into the

00:40:32 flame and burned. This instrument measures the amount of sodium and potassium in the

00:40:37 blood. Each element gives off a different color, and then the amount is recorded on

00:40:43 this meter here, which tells the doctor exactly how much sodium and potassium in your blood,

00:40:47 which is very helpful for him in diagnosing your problems.

00:40:50 Well, I can see how this instrument then would be extremely helpful to the medical profession.

00:40:54 Jack, you know one thing. It seems to me the names you get for these devices are enough

00:40:58 to scare some people off. For instance, you refer to this as a spectrophotometer. Isn't

00:41:02 there a simpler name for that?

00:41:04 Well, Ken, I think you might call it a fingerprint machine, if you like.

00:41:08 Well, that sounds a lot better. Why would you call it that, Jack?

00:41:10 Well, you know how the FBI takes your fingerprint to tell exactly who you are?

00:41:14 Mm-hmm.

00:41:15 The chemist uses this machine to tell exactly what chemicals are present in a mixture and

00:41:20 exactly how much of them. The instrument draws a fingerprint, if you like. You can see it

00:41:26 taking place here, and the chemist calls this a spectrum. And by looking at it, he can tell

00:41:30 exactly what chemicals are present and how much.

00:41:33 Well, what different uses would this have?

00:41:35 Oh, for example, a chemist working for a cigarette company can tell exactly how much

00:41:39 nicotine there is in tobacco. Or in the steel industry, how much chromium in steel. Or,

00:41:47 for example, in the pharmaceutical industry, they tell how much vitamin A is present in

00:41:51 pills. In fact, vitamin A was synthesized on this machine, Ken.

00:41:55 Now, this machine on the end here is one that certainly intrigues me, Jack. What is this

00:41:59 called?

00:42:00 This is a 200-channel automatic recorder, Ken. It's one of our industrial products. And

00:42:05 this instrument has been built for the Douglas Aircraft Company. It will automatically record

00:42:12 200 strain gauges on a wing to measure the stresses in the wing. It does it real rapidly

00:42:17 and automatically. Would you like to see how it works?

00:42:19 We certainly would, Jack.

00:42:20 Here, I'll start it up. And as you can see, it's recording automatically each of the

00:42:25 different instruments on a separate card. This used to take many, many man-hours to

00:42:29 do as accurately as this machine does automatically.

00:42:34 Well, Jack, I certainly wouldn't say that we completely understand these feats of

00:42:39 electronic wizardry, but I think we've all got now a very healthy respect for the

00:42:44 important job that they do.

00:42:47 The thermocouple is really a special kind of thermometer used to measure very small

00:42:51 amounts of heat in a Beckman spectrophotometer. Some of the parts which make up the

00:42:55 thermocouple are so small that a microscope must be used to assemble them. Here are the

00:43:00 parts that must be assembled inside the thermocouple tube.

00:43:04 We are now looking at Mrs. Orenson, a highly skilled Beckman employee whose intricate job

00:43:09 is to assemble and solder the component parts of the thermocouple. Let's join Ken Peters

00:43:14 as he finds out more about this stage of assembly.

00:43:19 Mrs. Orenson, do you enjoy working with these tiny parts?

00:43:22 Oh, yes, very much. In fact, I find it a real challenge.

00:43:25 Well, now I understand this is receiver assembly that you're going to make here.

00:43:29 Could you show us the steps involved in making it?

00:43:31 Yes, we'll take the small particle of gold and place it on the plate.

00:43:35 You mean those tiny little specks we saw on that glass are the actual receivers?

00:43:39 Yes.

00:43:40 And you're going to weld wires onto that?

00:43:42 A small wire will be welded to the gold.

00:43:44 The wires are in there? We'll have to take your word for it because they're so tiny that

00:43:48 we can't see them.

00:43:50 I'll place it on the plate.

00:43:52 Then how do you manipulate them around on the plate into the right position?

00:43:55 With the one-haired brush.

00:43:57 A one-haired...

00:43:58 Yes.

00:44:00 Yes, there is one hair there.

00:44:02 Is that any special kind of hair?

00:44:04 Oh, yes, it's quite special hair.

00:44:05 In fact, we get it from the heads of our distinguished guests and visitors,

00:44:08 and we'd like to have one of yours, Mr. Peters.

00:44:10 I guess I could spare one for science.

00:44:13 And now this is a little spot welding unit in which you will actually spot weld.

00:44:17 I imagine if you sneezed here, you'd be out of business for quite a little while,

00:44:20 wouldn't you?

00:44:21 Oh, yes.

00:44:22 In fact, we did have a sign in here that we shouldn't breathe in this room.

00:44:26 No breathing allowed.

00:44:27 Now I have the wire in place and the electrode on it,

00:44:30 and all I have to do to weld it is to press this little button and pray.

00:44:34 Mm-hmm.

00:44:37 There it is.

00:44:38 Now look.

00:44:39 You might see it.

00:44:40 Beautiful job, I'm sure.

00:44:42 How long have you been doing this type of work?

00:44:44 Well, I've been with Beckman about 12 years,

00:44:46 but I've been doing this work about five years.

00:44:49 Doesn't it make you nervous?

00:44:50 Oh, not at all.

00:44:51 In fact, the successful results are a great feeling of achievement.

00:44:56 You never feel like going home and beating your husband, then?

00:44:58 Oh, not for this.

00:45:00 Well, thank you very much for letting us interrupt you, Mr. Doretson.

00:45:03 You're welcome.

00:45:04 Thank you.

00:45:11 Well, aren't you Beckman?

00:45:20 Well, you've often asked yourself, why should we have a history of chemistry?

00:45:26 Many reasons for it, of course.

00:45:28 We do put things in perspective and it does enable us to come back

00:45:32 and remember things in the past.

00:45:35 I've seen this film here, Jack Bishop.

00:45:37 You know where the film went?

00:45:39 Well, you may wonder how we get our employees for Beckman Institute.

00:45:43 We got Jack Bishop.

00:45:45 He walked in my office one day.

00:45:47 We were in South Tuscaloosa.

00:45:48 He was a graduate.

00:45:49 He got an MBA degree from Harvard Business School.

00:45:52 It turned out that Beckman Institute was one of the companies that was used there as a case study.

00:45:58 He said, he'd become interested in what we made.

00:46:00 He'd like to go to work for us.

00:46:02 No more to do that.

00:46:03 He was hired in a spot.

00:46:04 He became one of our very, very best managers over the years.

00:46:09 Althea Orenson.

00:46:11 She had to leave and move to Hawaii when her husband went out there.

00:46:18 So she came in with tears in her eyes.

00:46:20 For 24 years, she had been doing nothing but painting with black,

00:46:26 with a one-haired brush in her hand.

00:46:29 I said, look, Beckman.

00:46:32 There are many rewards for being in the chemical industry.

00:46:38 Before that, let me talk again about perspective.

00:46:42 Back in the early 1930s, the 20s and 30s,

00:46:46 the Cal Tech couldn't be in the chemistry department.

00:46:50 Arthur Amos Noy was the head of the chemistry department then.

00:46:54 He would have nothing to do with commercialization.

00:47:01 When I started making the pH meter at 35,

00:47:05 I would make them just a few at a time over weekends.

00:47:09 Finally, by 39, the business had grown to the size that somebody had to run it full time.

00:47:15 I had to make a very difficult decision.

00:47:18 Should I leave the position there,

00:47:20 which I loved, of being an assistant professor at Cal Tech,

00:47:24 doing research and enjoying the teaching,

00:47:27 and engage in this crass commercialism of making a business out of this thing,

00:47:33 or not?

00:47:34 Well, by that time, I'd been enjoying some of the business aspects

00:47:39 as well as the scientific aspects,

00:47:41 so I decided to make the change.

00:47:44 I resigned from Cal Tech at that time.

00:47:47 I've been able to maintain my connections with it, of course, over the years,

00:47:51 and I'm the board of directors,

00:47:53 and chairman of the American Board of Trustees now,

00:47:55 so I still maintain a very close relationship.

00:48:02 A mention was made about the high cost of instrumentation,

00:48:05 so I looked back, and let me go.

00:48:08 May I give them some advice?

00:48:12 You who are students on here,

00:48:14 be prepared for almost anything that you go ahead in life,

00:48:18 because you never know what's going to come up.

00:48:20 I started out to be a chemical engineer.

00:48:23 By the time I got around to my doctorate degree,

00:48:26 I was a physical chemist,

00:48:27 and I studied the quantum yields and the photochemical reactions.

00:48:31 By the time I stepped from chemical engineering,

00:48:34 I ended up making instruments.

00:48:37 Well, on my research,

00:48:40 I worked, among other things, on the photochemical decomposition of hydrazine,

00:48:44 N2H4.

00:48:45 Believe it or not, that was not commercially available,

00:48:48 and I had spent weeks making a few milliliters of hydrazine.

00:48:53 If someone had said then,

00:48:55 in time coming, you can buy this stuff with a tank carload,

00:48:58 it's going to be used for hoisting spacecraft into space,

00:49:01 I thought it was out of his mind,

00:49:02 yet that's what was the reality of it.

00:49:05 So, as you look ahead, be prepared for anything,

00:49:09 whatever happens will exceed your wildest imaginations, I think, right now.

00:49:15 As I look back, there are many rewards, I'd say, of being in the business.

00:49:20 One is that it is gratifying to see people like you

00:49:25 making use of instruments to advance mankind,

00:49:28 and I'm happy to have a little part in making the tools that are useful to you.

00:49:34 It also is very comforting to have employees,

00:49:37 such as Jack Bishop, Alva Orson.

00:49:39 We pride ourselves.

00:49:41 We have the longest service in more than any company I've ever known.

00:49:49 It's very gratifying when I have employees come up and say,

00:49:51 well, thanks for providing me a job,

00:49:54 and maybe we can put my kids through school.

00:49:56 We have second and third generation employees coming through.

00:49:59 That's a very, very satisfying factor, indeed.

00:50:05 Finalizing ahead, we're making many of the newer instruments now.

00:50:11 Automatic instrumentation is becoming more and more important.

00:50:14 We make automatic protein and peptide sequences, as you know,

00:50:20 and synthesize it, DNA, synthesize it.

00:50:30 The human genome is going to be decoded one of these days,

00:50:33 and that's going to call for automatic equipment,

00:50:36 because we just don't have the manpower or the money to do that.

00:50:39 So one of our hottest items now is a robot.

00:50:42 It carries on all the operations you normally do.

00:50:45 It'll carry on filtration, separation, all these things entirely automatically.

00:50:50 And Dr. Lee Wood, Dr. Phil Tick, and others are very excited about this

00:50:55 and being one way to go to make the structure of the human genome become a reality.

00:51:03 We can carry it out, but we couldn't automate the process enough,

00:51:06 so we don't need this intense human factor component.

00:51:14 Well, aren't I supposed to say any more now, huh?

00:51:17 Thank you.

00:51:36 Well, we are just a touch behind schedule.

00:51:42 And we do have a break, and we think we should keep it,

00:51:45 but if you could hold it for ten minutes and come back, that would be very, very nice.

00:51:49 And thank you very much.