Pathways of a Cell Biologist: Through Yet Another Eye

280 Pages·2016·31.8 MB·English

by  Shinya Inoué (auth.)

Springer Biographies
Pathways of
a Cell Biologist
Through Yet Another Eye
SHINYA INOUÉ
Springer Biographies
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Shinya Inoué
Pathways of a Cell Biologist
Through Yet Another Eye
Shinya Inoué
Marine Biological Laboratory
Woods Hole , MA , USA
Springer Biographies
ISBN 978-981-10-0946-4 ISBN 978-981-10-0947-1 (eBook)
DOI 10.1007/978-981-10-0947-1
Library of Congress Control Number: 2016949742
© Springer Science+Business Media Singapore 2016
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Cover caption: Shinya investigates living cells with his polarized light microscope
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Foreword
Having had the honor of receiving the 2008 Nobel Prize in Chemistry, I, Osamu
Shimomura, am now being presented the additional honor to prepare a foreword for
Prof. Shinya Inoué’s autobiography P athways of a Cell Biologist . Although having
to rely on his autobiography as far as his personal life, especially as related to his
youth, I am more or less familiar with Shinya’s contribution to science, especially
since we have been coworkers at the Marine Biological Laboratory (MBL) in Woods
Hole for some 34 years.
Born in London, England, in 1921 and raised in China (Quemoy); Portland,
Oregon; and Sydney, Australia, Shinya’s early language was primarily English
especially since his Japanese diplomatic parents had not insisted on their use of the
Japanese language at home. In fact Shinya’s English was so familiar to him that he
won fi rst prize in his English composition class after arriving in Sydney after
Portland. And after returning to Tokyo from Sydney, he had to catch up on his
Japanese language quickly enough so that he had to repeat the 5th grade in the
Magome primary school rather than enter the 6th grade in order to adequately pre-
pare for the competitive admission to high school. Fortunately he had a jovial,
understanding teacher at Magome and was admitted to the 2nd city high school in
Ueno, Tokyo.
Not only was Shinya’s early upbringing abroad in these English-speaking coun-
tries conducive to his interest in the natural sciences, but especially his distaste for
high-school-taught Japanese history must have had clear infl uences on his career
choice. One main saving aspect of his high school experience was the extracurricu-
lar shop class offered by an interested graduate. There Shinya delighted in learning
to design and make vacuum-tube radios, amplifi ers, steam engines, and other
devices extracurricularly.
Shinya’s interest in science, especially in deciphering the mechanism of cell divi-
sion in live cells, became strongly motivated in 1941, when he was introduced to
Katsuma Dan (Katy, a nickname used by himself) at Musashi Koutou Gakkou
(Japanese senior high school). Katy had acquired his Ph.D. 10 years previously at
the University of Pennsylvania under L.V. Heilbrunn, where he met Jean Clark, also
a Ph.D., who was to be his wife. Despite this international hostile background,
v
vi Foreword
Shinya and Dan became lifelong friends who collaborated on their biophysical stud-
ies of living cells over many years.
Shinya and Katy found (following W. J. Schmidt in Tubingen) the special advan-
tage that polarized light microscopy provided for studying dynamic molecular
events in living cells. Early on they (S. & K.) demonstrated how the submicroscopic
fi bers that made up the spindle fi bers (which otherwise were not visible without fi x-
ing the cells) could be visualized with the polarizing microscope. Then in 1948 Katy
and Jean managed a fellowship for Shinya to enter the graduate school at Princeton,
New Jersey. Once at Princeton, Shinya’s mentor, Kenneth Cooper, encouraged
Shinya to follow his own interest in pursuing his thesis research.
Thus Shinya fi rst studied changes in the birefringence of the mitotic spindle
associated with cell division while concurrently adding some improvements to the
polarizing microscope which led to his Ph.D. thesis. Based on those studies, he
proposed how the rearrangement of the spindle molecules generated the forces that
pulled chromosomes apart. These studies were backed by studies on the infl uence
of colchicine on the morphology and birefringence of metaphase-arrested
Chaetopterus spindles as well as the effects of temperature on the morphology and
birefringence of the C haetopterus developing meiotic spindles.
A dramatic improvement in the sensitivity and resolving power of the polarizing
microscope was introduced a few years later by Shinya and coworkers at the
American Optical Company Research Group in Southbridge, Massachusetts. What
was known up to then was that even using objective and condenser lenses com-
pletely free of strain birefringence, the numerical aperture of the polarizing micro-
scope had to be quite limited by reducing the opening of the condenser aperture in
order to maintain a reasonably high extinction factor. But closing down the con-
denser aperture not only reduced the amount of light available through the micro-
scope but limits its resolving power drastically.
W hat Shinya had known before his seminar presentation at Southbridge was that
the back aperture of the polarizing microscope was only uniformly and nearly com-
pletely dark between crossed polarizers when the condenser aperture was quite
closed down, say to a numerical aperture (NA) of approximately 0.3. At any larger
condenser NA, the back aperture between crossed polarizers no longer was very
dark but became a dark center surrounded by 4 quadrants of increasing brightness
away from the center. Between the quadrants were arms of a dark cross paralleling
the polarizer- and analyzer-crossed orientation.
When the polarizer and analyzer were not set at exactly 90° to each other, the
dark arms of the cross opened up into two Vs in opposite quadrants. The newly
formed dark Vs indicated regions of the back aperture which were now extinguished
by the analyzer. And it took some 8 or 9 ° of analyzer rotation for the two Vs to reach
the edge of the back aperture of a 1.25 NA condenser and objective lenses. In other
words, the spherical lens surfaces of the condenser and objective lenses had induced
that much rotation away from the 90 ° crossed angle set by the polarizer and ana-
lyzer. No wonder the extinction factor was drastically reduced as one opened the
condenser diaphragm and let higher NA beams through.
Foreword vii
As proposed by Lem Hyde of Nikon, a half-wave birefringent plate should
reverse the rotation induced by the curved surfaces of the condenser and objective
lenses. Therefore why not introduce a half-wave plate between the condenser and
objective lenses and let the two rotations cancel each other out? As noted by Shinya,
the half-wave plate between the condenser and objective would indeed change the
rotation of light at the lens surfaces according to the curvature of each point that
light entered or exited the glass surface; hence rotation by the lens surfaces in the
condenser and objective would complicate but not cancel each other away.
Shinya’s solution was to introduce the half-wave plate at the aperture plane
(where the rays were now nearly parallel) outside of the condenser or objective.
This gave rise to the polarization rectifi er which in fact raised the extinction factor
of a pair of 1.25 NA lenses by 20-fold, incredibly several times better than the fi ve-
fold improvement required by Dr. Brian O’Brein of the University of Rochester
Optics Institute in order for American Optical to support Shinya’s research to
improve the extinction factor of the polarizing microscope!
W ith the extinction factor thus raised, and especially effectively so at high NAs,
the imaging and birefringence-detecting sensitivity of the polarizing microscope
were dramatically improved. Not only were the weakly birefringent microtubules in
the cytoplasm clearly imaged, but the detailed patterns of the weakly birefringent
fi bers contouring the surface of epithelial cells now stood out for the fi rst time. At
the same time, the detailed arrangement of DNA molecules in maturing spermato-
cytes became analyzable for the fi rst time.
T hus Shinya was able to acquire a series of birefringent images of maturing
spermatocytes of the crane fl y and from their compensated images to map out the
detailed arrangement of the intracellular DNA. This led to the demonstration of the
double helical structure of the maturing chromosomes, which themselves were
clearly delineated along the length of the maturing spermatocytes. The intricate
three-dimensional arrangement of the chromosomes thus revealed with the rectifi ed
polarizing microscope required their high-resolution birefringence analytical capa-
bility, but at such dimensions that the same chromosomes could not be visualized by
electron microscopy.
In addition to the barrier-breaking fi ndings based on the use of the rectifi ers in
the polarizing microscope, additional discoveries were made by Shinya and col-
leagues by further improvement of the birefringent-detecting sensitivity of the
polarizing microscope by combining it with TV imaging and electronic image pro-
cessing. For these developments, the physiology course at the Marine Biological
Laboratory in Woods Hole played important roles. In 1963, Shinya and Bob Allen
gave their fi rst physiology course presentations which, together with the faculty,
they had assembled for those occasions. During the course demonstrations, both
Bob and Shinya and their faculty found the surprising improvement that image con-
trast could be vastly improved by passing the image output of the microscope
through a TV system.
I n more recent advances, we must pay special tribute to the contribution of many
specialists from optical, electronic, and computer companies and to the new physi-
cists specializing in electronics. Their contributions made by many specialists with
viii Foreword
a variety of physical backgrounds have not only opened the eyes of other scientists
but especially have contributed to those primarily interested in biology.
A mong the several honors and awards received by Shinya for his professional
contributions [Ju-shou Oyobi Eiyo in Appendix 4 of Through Yet Another Eye ],
Japanese readers may be especially interested in the International Prize for Biology
that was awarded to Shinya in 2003 by the Japan Society for the Promotion of
Science. For this occasion, Shinya and Sylvia were able to bring most of the Inoué
family from the United States to Japan and have a reunion with their Japanese rela-
tives. Together they also attended the award ceremony at which the Emperor and
Empress of Japan offi ciated.
I n his speech given at the ceremony, the Showa Emperor fi rst congratulated
Shinya for his achievements and contribution to science. But as recorded in
Appendix 2 of T hrough Yet Another Eye, the Emperor’s recollection included how
Shinya started his research overcoming diffi cult wartime problems. That recollec-
tion was especially touching for Shinya, who had in fact gone through those trying
days. Both the Showa Emperor’s speech and Shinya’s response at this ceremony (in
Appendix 3) can be found under “International Prize for Biology” in Chap. 5 .
The balance of his autobiography describes the excitement that Shinya, his stu-
dents, and colleagues experienced as they further explored the behavior of mole-
cules and their assemblies, which underlie the behavior of living tissues and
organisms. Fortunately they continued to unravel hidden secrets of living cells and
organisms while exposing the excitement associated with discovery by new bio-
physical means. Hopefully, their research will resonate among readers and encour-
age them to explore similar paths of scientifi c exploration.
As to the interaction of my family with Shinya’s, they tended to be private. When
in the fall of 1981, we decided to move from Princeton to the Marine Biological
Laboratory in Woods Hole, my family remembers how Dr. and Mrs. Woody Hastings
invited the Inoués to join the dinner party at their home. This was our fi rst interac-
tion with the Inoués. The Inoués’ youngest son, Ted, was about the same age as our
own youngest son, Tsutomu, and both being interested in computers were friendly
with each other until they entered the university. Our daughter Sachi had a summer
job in Shinya’s laboratory, and we recall how she told us about learning about feed-
ing the sea urchins and how to carefully prepare the glassware in which to grow the
urchin larvae.
In 2002, I retired from the Marine Biological Laboratory in Woods Hole myself.
On that occasion, Shinya arranged a memorial symposium in my honor. With little
knowledge on my part, Shinya had assembled those scientists interested in aequorin
and green fl uorescent proteins (GFPs). Those scientists included Roger T’sien from
California, with whom I shortly shared the Nobel Prize in Chemistry. The audience
fi lled the available seats and surprised me with the breadth of Shinya’s scholastic
knowledge and acquaintances. At this meeting, Prof. Ford from Rutgers University,
who a couple of years later in 2004 co-organized the Friday Harbor Lab 100th
Anniversary Symposium, also joined us. The topic of the symposium was “calcium-
dependent bioluminescent protein and green fl uorescent protein.” Shinya was pres-
ent at this symposium and reported up-to-date information on the fl uorescent
Foreword ix
polarization of GFP protein crystals, which I had purifi ed for his polarized light
microscopy. For this meeting at the Friday Harbor Labs, Shinya’s family and we
occupied adjoining cottages. And I recall how Mrs. Inoué, who had been raised in
the state of Washington, had thoroughly enjoyed the island by driving her car to
various points of interest there. That same summer of 2004, a memorial dinner was
held at the Marine Biological Laboratory in Woods Hole to commemorate Shinya’s
contributions to science, and we too were invited for the occasion. Shinya’s friends
from long distances joined the occasion, which was most appropriate considering
his having received the International Prize for Biology the previous year. At this
meeting, I remember the many representatives of the world’s top microscope com-
panies who recalled the severe demands from Shinya, who challenged them with
improvements of the microscopes which they were barely able to satisfy by exercis-
ing their utmost efforts.
Since those days, we met on occasions when Shinya was the recipient of various
prizes and on the occasion of me receiving my Nobel Prize. With each of these occa-
sions, I have been impressed by how many individuals around the world loved
Shinya as a humanistic researcher. As described in his autobiography, not only those
in Japan and the United States but research collaborators appear from around the
globe. To me, this is an indication not only of Shinya’s clear observing power and
sharp intellect but that of his human attractiveness. I am confi dent that the strength
of character of Shinya refl ected in this autobiography will further benefi t the char-
acter of international students who digest this essay.
Woods Hole, MA, USA Osamu Shimomura
August 2015