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Henry Bourne
Forum
TRENDS in Cell Biology Vol.12 No.4 April 2002
201
Profile
Fig. 1. Henry Bourne.
differ very much from one another. That difference actually makes science work. If scientists were all alike, science would grind to a halt. Gordon Tomkins was enormously imaginative and always excited about any new question. Unlike most of us, when Gordon had an idea that turned out to be wrong, he was delighted, because he had plenty of other ideas and didn’t have to worry about that one any more. He felt that new ideas should be tested, but, if they are wrong, then you go on to do something else. His approach really changed my view of the way science should be done.
Why did you stay in research rather than go back to a being a physician?
Which of your accomplishments are you particularly proud of?
It was the idea that science always gives you something new. With a clinical career, it is exciting at the beginning because everything is new, but, once you learn your profession, the problems keep reappearing. In science, the questions just keep on coming. Something is different each year, each week. It took me years to realize this, but fortunately I also had my stethoscope stolen at just the right time to push me over the edge.
This is a hard question. It’s not an accomplishment, and it’s a funny thing to be proud of. But it is a fact that science has turned out to be so much fun. When it comes down to it, we all care about accomplishing things, but many people feel that they are working very hard, and worry whether or not that they are making a huge contribution. I’ve not worried a lot about that, but instead just been grateful to enjoy science.
Henry Bourne A physician by training, Henry Bourne (Fig. 1) first joined the US National Institutes of Health in an attempt to evade the Vietnam war. At the NIH, he became hooked by science and went on to a fellowship at the University of California, San Francisco (UCSF). Bourne became a researcher and instructor at UCSF in 1971 and is now professor of pharmacology there. His lab was one of the first to investigate signaling by trimeric G proteins and, in recent years, has focused on the cellular mechanisms in chemotaxis. What event led you into research?
I was a doctor and about to be drafted to the war in Vietnam. One way out of this was to work at the National Institutes of Health [NIH]. I had always had the inclination to do some science, and I managed to wangle my way into the NIH. There, I discovered how exciting science could be. After a couple of years at the NIH, I did a fellowship at the University of California, San Francisco, and then became a faculty member. I didn’t do a PhD; in essence, my PhD was the two years at the NIH and my years as an assistant professor. In those days, fortunately, you didn’t have to hit the ground running. Instead, I crawled and stumbled, and the system gave me enough slack to learn how to become a scientist. How did you become interested in your area of research?
I sort of slid into it by virtue of the fact that I was working on cyclic AMP, which back in those days was a ‘sexy’ thing to work on. I began to focus on G protein research when I met Gordon Tomkins. Tomkins, who was my real scientific hero, showed me it was possible to do somatic genetics in mammalian cells. We isolated a mutant cell line that Elliott Ross and Al Gilman used as a tool to discover the first G protein, Gs. I isolated the mutant in 1975, and their discovery was made soon afterwards. They beat me to the finish line by many miles, but their unraveling of the Gs story turned out to be one of the best things that ever happened to me. This was the beginning of the G protein world. I was hooked! http://tcb.trends.com
What would you like to see your lab accomplish in the next 5–10 years?
We are trying to shed some light on how a cell knows where to go when it is crawling towards an attractant chemical — that is, in chemotaxis. We are trying to figure out how a cell interprets the external gradient of attractant. Many cells can interpret differences in concentrations that are as small as a couple of percent from one end of the cell to the other. The general principles are poorly understood. We have a rudimentary parts list, but we don’t know how they fit together. My hope is that the next decade will give us real insight into how this extraordinary process works. What do you think distinguishes an excellent from a good scientist and why?
There’s an infinite number of ways to be excellent. Most scientists are very smart and capable, so a lot depends on luck, and of course on imagination and ambition. There is not one single way to be an excellent scientist. If you consider Nobel Prize winners and other scientists you admire, their attitudes and styles often
…everyone in my lab who has gone on to a ‘… successful career in science genuinely loved doing experiments.’
Do you have any advice for graduate students or postdocs who are just starting out?
They should enjoy it. And if they don’t, they should be doing something else. That doesn’t mean they should enjoy every minute. There’s a lot of hard slogging and disappointment. But, if it doesn’t fundamentally give you a thrill to be thinking about science and doing experiments, then you ought to get out quickly. I hear a lot of postdocs and graduate students who are worried about the state of the economy and future support for science. But the fact is that science has never been more exciting than it is now. An industrialized country such as ours will support science for a long time. It’s not a way to get enormously rich, but it is a way to guarantee that what you are doing five years from now will be different from what you are doing now.
0962-8924/02/$ – see front matter © 2002 Elsevier Science Ltd. All rights reserved. PII: S0962-8924(02)02264-X
202
Forum
TRENDS in Cell Biology Vol.12 No.4 April 2002
Advice is cheap, of course, and maybe I should point out one of my character defects, which I hope students won’t suffer from: I have always been more interested in the ideas and the results of science than in doing the actual experiments. You can still get a lot out of science by caring about other people’s experiments, of course. I
like to think about things, talk to people, and get the right experiment done one way or another. That’s not the best way, I know. In fact, everyone in my lab who has gone on to a successful career in science genuinely loved doing experiments. I envy that enormously, in the same way that I envy people who sing or play an
Henry Bourne was interviewed for BioMedNet by Emma Hitt. Adapted from a ‘Conference Reporter’ feature published online on BioMedNet (http://news.bmn.com/conferences/).
Function is a new addition to this array, although it has a slightly different scope, trying to cover both fundamental principles and techniques, as well as clinical applications and diagnostic approaches. For this purpose, selected experts in the fields contribute various chapters, adding to those produced by the editors. The book is clearly aimed at the novice in flow cytometry and starts off with six chapters discussing general principles, hardware, sample preparation, fluorescent labels, data analysis and quality control. Although the chapters are structured in a similar manner, aiming to introduce a certain homogeneity, the coverage of the different topics is rather variable. The two chapters on sample preparation and fluorochromes are very detailed and highly informative, applicable also for the non-clinical scientist. The first chapter provides useful practical protocols on human leukocyte isolation and fixation, as well as the necessary hints and technical tricks to avoid common pitfalls. The second gives a detailed theoretical background on the physics of fluorescence, as well as an exhaustive list of different fluorescent labels and their application in different staining techniques. By contrast, the introductory chapter on general principles of flow cytometry is not presented at the same level, perhaps because the author (M.G. Macey) wanted to keep it as ‘simple’ as possible. Besides containing some out-dated information (especially concerning the hardware of different manufacturers), some of the explanations are not sufficiently detailed, and especially the issue of fluorescent spectral compensation should have been more clearly explained. Spectral compensation certainly causes a lot of confusion for newcomers, but
unfortunately the figures demonstrating the effects of electronic compensation are actually examples on how ‘not’ to compensate. More appropriate are the chapters on quality control and data analysis; both are concise, providing the necessary information without going into too much detail, and with an extensive list of references. A special chapter has been reserved for clinical use of laser-scanning cytometry, a technology that is a ‘hybrid’ between cytometry and confocal microscopy, which allows correlation of immunophenotypes, conventional histological staining and in situ hybridization data for individual cells. Richly illustrated with color figures, the reader is guided through a practical example using cell relocalization for sequential light microscopy, immunophenotyping and in situ hybridization. The second part of the book introduces several techniques used in clinical research, starting with a very didactic chapter on ‘Leukocyte immunobiology’, followed by ‘Immunophenotyping in disease’. Both provide a good introduction into clinical diagnostics and are backed up by appropriate references for those requiring further details. ‘Analysis and isolation of minor cell populations’ covers a more specialist area, discussing enumeration of reticulocytes, as well as quantification of fetal–maternal hemorrhage. While the first part of this chapter is very informative, the section on cell-sorting strategies and the respective ‘statistics’ is not really relevant for a beginner, given that high-speed cell sorters still require specially trained personnel. Cell viability, necrosis and apoptosis, as well as cell-cycle and DNA analysis are all quickly overviewed, with short protocols for the most frequently used techniques. However, a more extensive list of
instrument. But you have to play the cards you’re dealt… and hope for the best.
Book Review
Going with the flow – a beginners guide to an exciting technology Cytometric Analysis of Cell Phenotype and Function Edited by Desmond A. McCarthy and Marion G. Macey. Cambridge University Press, 2001. £95.00 hbk (413 pages) ISBN 0 521 66029 7
‘Lamina flow’ – although a little ‘typo’ on the second page of Cytometric Analysis of Cell Phenotype and Function – describes well the early days of flow cytometry, when scientists used special slides to let cells to flow one by one through a microscope. Make them flow in a ‘laminar’ stream, allowing for precise excitation by a laser, and you have the prototype of a flow cytometer. Cells can be analyzed (and on some machines sorted) for their light scattering and up to 11 fluorescent labels can be used. Moreover, recent advances in high-speed computing, monoclonal antibody technology and more colorful fluorescent ‘XFP’ variants have made flow-cytometric analysis and cell sorting an ‘exciting’ technology, frequently used in modern immunology, cellular and molecular biology. Over the years, it has been also applied in clinical research and diagnostics, such that bench-top flow cytometers are already standard equipment in modern clinical laboratories. Several comprehensive books have been published about flow cytometry in general, although most of them are directed towards basic research. Cytometric Analysis of Cell Phenotype and http://tcb.trends.com
0962-8924/02/$ – see front matter © 2002 Elsevier Science Ltd. All rights reserved. PII: S0962-8924(01)02248-6
TRENDS in Cell Biology Vol.12 No.4 April 2002
201
Profile
Fig. 1. Henry Bourne.
differ very much from one another. That difference actually makes science work. If scientists were all alike, science would grind to a halt. Gordon Tomkins was enormously imaginative and always excited about any new question. Unlike most of us, when Gordon had an idea that turned out to be wrong, he was delighted, because he had plenty of other ideas and didn’t have to worry about that one any more. He felt that new ideas should be tested, but, if they are wrong, then you go on to do something else. His approach really changed my view of the way science should be done.
Why did you stay in research rather than go back to a being a physician?
Which of your accomplishments are you particularly proud of?
It was the idea that science always gives you something new. With a clinical career, it is exciting at the beginning because everything is new, but, once you learn your profession, the problems keep reappearing. In science, the questions just keep on coming. Something is different each year, each week. It took me years to realize this, but fortunately I also had my stethoscope stolen at just the right time to push me over the edge.
This is a hard question. It’s not an accomplishment, and it’s a funny thing to be proud of. But it is a fact that science has turned out to be so much fun. When it comes down to it, we all care about accomplishing things, but many people feel that they are working very hard, and worry whether or not that they are making a huge contribution. I’ve not worried a lot about that, but instead just been grateful to enjoy science.
Henry Bourne A physician by training, Henry Bourne (Fig. 1) first joined the US National Institutes of Health in an attempt to evade the Vietnam war. At the NIH, he became hooked by science and went on to a fellowship at the University of California, San Francisco (UCSF). Bourne became a researcher and instructor at UCSF in 1971 and is now professor of pharmacology there. His lab was one of the first to investigate signaling by trimeric G proteins and, in recent years, has focused on the cellular mechanisms in chemotaxis. What event led you into research?
I was a doctor and about to be drafted to the war in Vietnam. One way out of this was to work at the National Institutes of Health [NIH]. I had always had the inclination to do some science, and I managed to wangle my way into the NIH. There, I discovered how exciting science could be. After a couple of years at the NIH, I did a fellowship at the University of California, San Francisco, and then became a faculty member. I didn’t do a PhD; in essence, my PhD was the two years at the NIH and my years as an assistant professor. In those days, fortunately, you didn’t have to hit the ground running. Instead, I crawled and stumbled, and the system gave me enough slack to learn how to become a scientist. How did you become interested in your area of research?
I sort of slid into it by virtue of the fact that I was working on cyclic AMP, which back in those days was a ‘sexy’ thing to work on. I began to focus on G protein research when I met Gordon Tomkins. Tomkins, who was my real scientific hero, showed me it was possible to do somatic genetics in mammalian cells. We isolated a mutant cell line that Elliott Ross and Al Gilman used as a tool to discover the first G protein, Gs. I isolated the mutant in 1975, and their discovery was made soon afterwards. They beat me to the finish line by many miles, but their unraveling of the Gs story turned out to be one of the best things that ever happened to me. This was the beginning of the G protein world. I was hooked! http://tcb.trends.com
What would you like to see your lab accomplish in the next 5–10 years?
We are trying to shed some light on how a cell knows where to go when it is crawling towards an attractant chemical — that is, in chemotaxis. We are trying to figure out how a cell interprets the external gradient of attractant. Many cells can interpret differences in concentrations that are as small as a couple of percent from one end of the cell to the other. The general principles are poorly understood. We have a rudimentary parts list, but we don’t know how they fit together. My hope is that the next decade will give us real insight into how this extraordinary process works. What do you think distinguishes an excellent from a good scientist and why?
There’s an infinite number of ways to be excellent. Most scientists are very smart and capable, so a lot depends on luck, and of course on imagination and ambition. There is not one single way to be an excellent scientist. If you consider Nobel Prize winners and other scientists you admire, their attitudes and styles often
…everyone in my lab who has gone on to a ‘… successful career in science genuinely loved doing experiments.’
Do you have any advice for graduate students or postdocs who are just starting out?
They should enjoy it. And if they don’t, they should be doing something else. That doesn’t mean they should enjoy every minute. There’s a lot of hard slogging and disappointment. But, if it doesn’t fundamentally give you a thrill to be thinking about science and doing experiments, then you ought to get out quickly. I hear a lot of postdocs and graduate students who are worried about the state of the economy and future support for science. But the fact is that science has never been more exciting than it is now. An industrialized country such as ours will support science for a long time. It’s not a way to get enormously rich, but it is a way to guarantee that what you are doing five years from now will be different from what you are doing now.
0962-8924/02/$ – see front matter © 2002 Elsevier Science Ltd. All rights reserved. PII: S0962-8924(02)02264-X
202
Forum
TRENDS in Cell Biology Vol.12 No.4 April 2002
Advice is cheap, of course, and maybe I should point out one of my character defects, which I hope students won’t suffer from: I have always been more interested in the ideas and the results of science than in doing the actual experiments. You can still get a lot out of science by caring about other people’s experiments, of course. I
like to think about things, talk to people, and get the right experiment done one way or another. That’s not the best way, I know. In fact, everyone in my lab who has gone on to a successful career in science genuinely loved doing experiments. I envy that enormously, in the same way that I envy people who sing or play an
Henry Bourne was interviewed for BioMedNet by Emma Hitt. Adapted from a ‘Conference Reporter’ feature published online on BioMedNet (http://news.bmn.com/conferences/).
Function is a new addition to this array, although it has a slightly different scope, trying to cover both fundamental principles and techniques, as well as clinical applications and diagnostic approaches. For this purpose, selected experts in the fields contribute various chapters, adding to those produced by the editors. The book is clearly aimed at the novice in flow cytometry and starts off with six chapters discussing general principles, hardware, sample preparation, fluorescent labels, data analysis and quality control. Although the chapters are structured in a similar manner, aiming to introduce a certain homogeneity, the coverage of the different topics is rather variable. The two chapters on sample preparation and fluorochromes are very detailed and highly informative, applicable also for the non-clinical scientist. The first chapter provides useful practical protocols on human leukocyte isolation and fixation, as well as the necessary hints and technical tricks to avoid common pitfalls. The second gives a detailed theoretical background on the physics of fluorescence, as well as an exhaustive list of different fluorescent labels and their application in different staining techniques. By contrast, the introductory chapter on general principles of flow cytometry is not presented at the same level, perhaps because the author (M.G. Macey) wanted to keep it as ‘simple’ as possible. Besides containing some out-dated information (especially concerning the hardware of different manufacturers), some of the explanations are not sufficiently detailed, and especially the issue of fluorescent spectral compensation should have been more clearly explained. Spectral compensation certainly causes a lot of confusion for newcomers, but
unfortunately the figures demonstrating the effects of electronic compensation are actually examples on how ‘not’ to compensate. More appropriate are the chapters on quality control and data analysis; both are concise, providing the necessary information without going into too much detail, and with an extensive list of references. A special chapter has been reserved for clinical use of laser-scanning cytometry, a technology that is a ‘hybrid’ between cytometry and confocal microscopy, which allows correlation of immunophenotypes, conventional histological staining and in situ hybridization data for individual cells. Richly illustrated with color figures, the reader is guided through a practical example using cell relocalization for sequential light microscopy, immunophenotyping and in situ hybridization. The second part of the book introduces several techniques used in clinical research, starting with a very didactic chapter on ‘Leukocyte immunobiology’, followed by ‘Immunophenotyping in disease’. Both provide a good introduction into clinical diagnostics and are backed up by appropriate references for those requiring further details. ‘Analysis and isolation of minor cell populations’ covers a more specialist area, discussing enumeration of reticulocytes, as well as quantification of fetal–maternal hemorrhage. While the first part of this chapter is very informative, the section on cell-sorting strategies and the respective ‘statistics’ is not really relevant for a beginner, given that high-speed cell sorters still require specially trained personnel. Cell viability, necrosis and apoptosis, as well as cell-cycle and DNA analysis are all quickly overviewed, with short protocols for the most frequently used techniques. However, a more extensive list of
instrument. But you have to play the cards you’re dealt… and hope for the best.
Book Review
Going with the flow – a beginners guide to an exciting technology Cytometric Analysis of Cell Phenotype and Function Edited by Desmond A. McCarthy and Marion G. Macey. Cambridge University Press, 2001. £95.00 hbk (413 pages) ISBN 0 521 66029 7
‘Lamina flow’ – although a little ‘typo’ on the second page of Cytometric Analysis of Cell Phenotype and Function – describes well the early days of flow cytometry, when scientists used special slides to let cells to flow one by one through a microscope. Make them flow in a ‘laminar’ stream, allowing for precise excitation by a laser, and you have the prototype of a flow cytometer. Cells can be analyzed (and on some machines sorted) for their light scattering and up to 11 fluorescent labels can be used. Moreover, recent advances in high-speed computing, monoclonal antibody technology and more colorful fluorescent ‘XFP’ variants have made flow-cytometric analysis and cell sorting an ‘exciting’ technology, frequently used in modern immunology, cellular and molecular biology. Over the years, it has been also applied in clinical research and diagnostics, such that bench-top flow cytometers are already standard equipment in modern clinical laboratories. Several comprehensive books have been published about flow cytometry in general, although most of them are directed towards basic research. Cytometric Analysis of Cell Phenotype and http://tcb.trends.com
0962-8924/02/$ – see front matter © 2002 Elsevier Science Ltd. All rights reserved. PII: S0962-8924(01)02248-6