Home Cytometry History Individual Histories Al Brunsting
Al Brunsting
- Please tell us about a significant event or moment in Cytometry that you experienced, in a lab, at a conference, or at an information gathering.
In 1970 I was very fortunate to be accepted into the Flow Cytometry Group of the Los Alamos Scientific Labs. At that time I was a graduate student of the University of New Mexico Physics Department in Albuquerque, NM. The Cytometry Group was headed by Dr. Marvin Van Dilla. My site advisor was Dr. Paul Mullaney.
This was a high-energy group of about 10 staff members, including P. F. Mullaney, M. J. Fulwyler, and J. A. Steinkamp. When I was there (June 1970 – May 1972) the field of flow cytometry was being pioneered in our group as well as at Stanford University, the IBM Research Center, and several other places. Specifically, the developing technologies for the flow cytometric measurements on single cells were Coulter particle sizing, whole cell fluorescence, and light scattering.
Each week included several presentations by a staff member or by one of the many visitors, such as M. L. Mendelsohn, L. L. Wheeless, and R. G. Sweet. The associated questions and answers where simulating to young graduate student like me.
Al Brunsting atop Sandia Crest near Albuquerque, NM, about the time he joined the Flow Cytometry Group at Los Alamos Scientific Labs, Los Alamos, NM, 1969.
Dr. Mullaney encouraged me to work with light scattering from single cells. While my apparatus was being built in our model shop I developed a light scattering simulation which included some unique aspects. Applied Optics accepted my first manuscript on that simulation as did the Optical Society of America for presentation at their 1971 annual meeting.
Dr. Mullaney and I traveled to Washington, DC, where OSA met and I was scheduled to deliver my first paper on my work while my site advisor looked on. The audience also included many other notables in the field of light scattering such as M. Kerker and J. P. Kratohvil from Clarkson College of Technology.
I was more than a little nervous so I wrote out all possible questions I could think of that might be asked of me. After my first presentation to a technical society, based on my work, the floor was opened for questions. Four questions were asked. I had anticipated three of them and gave learned sounding responses, as if I knew what I was talking about. My response on the fourth question was “That question really involves Dr. Mullaney’s contribution to this work and I’m sure he would like to respond.” Sure enough, he stood and gave an excellent response. I was very relieved after that presentation.
- What do you feel was your major contribution in the field of Cytometry?
My major contributions included these two:
1. Light scattering simulations that could be applied to measurements. My work was based on that of A. L. Aden and M. Kerker who generalized the work of G. Mie. Starting from Maxwell’s equations Mie correctly predicted the light scattering fluxes from arbitrarily sized spheres with arbitrary indices. Aden and Kerker added a spherical coating onto Mie’s sphere.
Dr. Mullaney recognized that this enhancement could be used to model single biological cells where the inner sphere represented the nucleus and the coating represented the cytoplasm. Of course single cells have morphologies that are not at all like coated spheres. The anticipation was that for certain cell types and osmotic conditions the coated sphere might be good enough to help us understand the observed light scattering measurements.
The model of Aden and Kerker was coded in FORTRAN 4 and run on some of the world’s fastest computers (at that time) at Los Alamos by me. During this process certain insights were recognized about how the radiant energy is distributed within the cell and how that relates to the various elements in the summation series of the theory. My simulation was used many times for a variety of cell types and conditions at Los Alamos and at other labs at that time.
2. Apparatus to measure light scattering. Based on suggestions made to me at one of our early informal group meetings, I designed a space to capture scattered light. It consisted of a 55-gallon drum of which there were many at Los Alamos. A track was made on the inside where photographic film was located.
The film was calibrated, as was the whole densitometer using mono-disperse micro-beads, sized similar to the cells that were used. That apparatus yielded results that were similar to what the simulations predicted.
- What drove you to this achievement?
Whatever I achieved in this area was due in large part to the encouragement, knowledge, and patience of Dr. M. A. Van Dilla and especially Dr. P. F. Mullaney. There were many other people at Los Alamos and in the Physics Department of the University of News Mexico, especially Dr. Howard Bryant.
My dear wife, Joyce, taught school at this time and supported us. I was fortunate she was there. Our first child, David, was born in Los Alamos (1972).
- Why was it that your team was able to do it? Was it a special skill? Sudden insight?
- Los Alamos had enormous experience in the area of particle counting (including the associated electronics) and optics (including photo-multiplier tubes). Starting with the Manhattan Project Los Alamos had a “can do” and very inventive spirit when it came to technical challenges such a flow cytometry. When Wright Langham (M. A. Van Dilla reported to him) suggested a radically new direction (flow cytometry), M. J. Fulwyler built and successfully demonstrated a high speed cell sorter base on the Coulter principle. This was uncharted territory. But the people were talented and willing to persist until a series of challenging goals were met. Another important aspect was the group effort and sharing information with each other. It is hard to see how I could have landed in a better situation.
- How do you think your work impacted the field of Cytometry?
- When I arrived on the scene under the wing of Dr. P. F. Mullaney, there were but two types of measurements in Flow Cytometry: Single cell fluorescence and Coulter sizing of small particles. My work showed that certain aspects of cellular morphology could be determined from the light scattering patterns of single cells. This work also showed that this would be very challenging since the scattering formation was entangled in the scattered light. My work showed how some of this disentanglement might be carried out.
- Al Brunsting, Ph.D
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