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Biomechanical Energy Harvesting: Generating Electricity During

Walking with Minimal User Effort


J. M. Donelan et al.
Science 319, 807 (2008);
DOI: 10.1126/science.1149860

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REPORTS
study indicates 404, 0. F. J. F.
180 Van Veen, R.
that changes in (2000 J. Morris, H.
resource traits ). C. J. Godfray,
influence food Annu. Rev.
web diversity Entomol. 51,
and complexity 187 (2006).
by interacting 1. R. J.
Morris, O. T.
with foraging Lewis, H. C.
biology J. Godfray,
(indicated by Nature 428,
size-dependent 310 (2004).
2. A. P.
parasitism and Beckerman,
sex allocation) O. L. Petchey,
of consumers P. H. Warren,
across several Proc. Natl.
Acad. Sci.
trophic levels U.S.A. 103,
through a 13745 (2006).
cascade of 3. M.
density- and Omacini, E.
J. Chaneton,
trait-mediated C. M.
effects (Fig. 2), Ghersa, C.
with B. Muller,
implications for Nature 409,
78 (2001).
food web
4. N.
stability and Underwood,
ecosystem M. D.
functioning. Rausher,
Ecology 81,
References 1565 (2000).
and Notes 5. P. J.
1. M. D. Ode, Annu.
Hunter, P. W. Rev.
Price, Ecology Entomol. 51,
73, 724 (1992). 163 (2006).
2. O. J. 6. J. A.
Schmitz, P. A. Harvey, N.
Hambäck, A. P. M. Van
Beckerman, Dam, R.
Am. Nat. 155, Gols, J.
141 (2000). Anim. Ecol.
3. L. 72, 520
Oksanen, S. (2003).
D. Fretwell, 7. C.
J. Arruda, P. Gómez-
Niemalä, Campo, S.
Am. Nat. Prakash, in
118, 240 Developmen
(1981). ts in Plant
4. J. M. Genetics
Montoya, S. and
L. Pimm, R. Breeding, C.
V. Solé, Gomez-
Nature 442, Campo, Ed.
259 (2006). (Elsevier,
5. A. R. Amsterdam,
Ives, S. R. 1999), chap.
Carpenter, 4.
Science 317, 8. H. C.
58 (2007). J. Godfray,
6. K. S. Parasitoids:
McCann, Behavioral
Nature 405, and
228 (2000). Evolutionary
7. A. R. Ecology
Ives, B. J. (Princeton
Cardinale, Univ. Press,
Nature 429, Princeton,
174 (2004). NJ, ed. 1,
8. 1994), pp.
R. J. 9–10.
Willia 9. C. B.
ms, N. Müller, I. C. T.
D. Adriaanse, R.
Martin Belshaw, H. C.
ez, J. Godfray, J.
Nature Anim. Ecol.
68, 346 (1999). 0. F. www.science
10. D. Bersier, C. mag.org/cgi/c
J. Banasek- ontent/full/31
Sullivan, Richter, M. 9/5864/804/D
W. Völkl, F. Cattin, C1 Materials
Annu. Ecology 83, and Methods
Rev. 2394 S
Entomol. (2002). O
44, 291 1. F. J. M
(1999). F. van Veen,
11. J. M. C. B. T
Tylianakis, Müller, J. K. e
T. x
Pell, H. C. J.
Tscharntke, t
O. T. Lewis, Godfray, J.
Nature 445, Anim. Ecol.
77, 191 F
202 (2007). i
(2008).
2. The g
farm of s
Wageningen .
University
prepared the S
field layout. 1
G.
Bukovinszki t
ne Kiss and o
V. Taravel
helped in S
collecting 4
data. J. A. Table S1
Harvey 2
provided 5
feral J
Brassica u
seeds. Food l
webs were y
drawn with 2
code by H. 0
C. J. 0
Godfray. 7
;
M.D. and
a
T.B. were
c
funded by c
the e
Netherlands p
Organisation te
for Scientific d
Research– 2
Earth and 7
Life D
Sciences e
Council c
(NWO- e
ALW, VICI m
grant b
865.03.002) e
and F.J.F.vV. r
by the 2
Natural 0
Environment 0
Research 7
Council, 1
UK. Voucher 0
.1
specimens
1
were
2
deposited at 6
the /s
Laboratory ci
of e
Entomology n
(Wageninge c
n University, e.
reference 1
number 1
Buko2005.0 4
01). 8
3
Supporting 1
Online Material 0

Biomechanical Energy
Humans are a (COH), a
rich source of phone dimensionless
energy. An chargers (4). quantity
average-sized A limitation defined as the
person stores as of these con- addi-
much energy in ventional
fat as a 1000-kg methods is
battery (1, 2). that users
People use must focus
muscle to their attention
convert this on power
stored chemical generation at
energy into the expense
positive of other
mechanical activities,
work with peak typically
efficiencies of resulting in
about 25% (3). short bouts of
This work can generation.
be performed at For electrical
a high rate, power
with 100 W generation
easily over longer
sustainable (1). durations, it
Many devices would be
take advantage desirable to
of human harvest
Generating Electricity During Walking power capacity
to produce
energy from
everyday ac-
with Minimal User Effort electricity,
including hand-
tivities such
as walking.
crank It is a
]. M. Donelan,1* Q. Li,1 V. Naing,1 ]. A. Hoffer,1 D. ]. Weber,2 A. D. Kuo3 generators as challenge,
well as wind-up however, to
We have developed a biomechanical energy harvester that generates electricity during human flashlights, produce sub-
walking with little extra effort. Unlike conventional human-powered generators that use radios, and stantial
positive muscle work, our technology assists muscles in performing negative work, mobile electricity
analogous to regenerative braking in hybrid cars, where energy normally dissipated during from walking.
braking drives a generator instead. The energy harvester mounts at the knee and selectively
1
School of
Kinesiology, Most energy-
engages power generation at the end of the swing phase, thus assisting deceleration of the Simon Fraser harvesting
joint. Test subjects walking with one device on each leg produced an average of 5 watts of University (SFU), research has
electricity, which is about 10 times that of shoe-mounted devices. The cost of harvesting — Burnaby, BC V5A focused on
the additional metabolic power required to produce 1 watt of electricity —is less than one- 1S6,
2
Canada.
generating
Department of
eighth of that for conventional human power generation. Producing substantial electricity Physical Medicine electricity
with little extra effort makes this method well-suited for charging powered prosthetic limbs and Rehabilitation, from the
and other portable medical devices. University of compression
Pittsburgh,
Pittsburgh, PA of the shoe
15213, USA. sole, with the
3
Departments of best devices
Mechanical generating 0.8
Engineering and
Biomedical
W (4). A
Engineering, noteworthy
University of departure is a
Michigan, Ann spring-loaded
Arbor, MI 48109,
USA.
backpack (5)
that harnesses
*To whom
correspondence the vertical
should be oscillations of
addressed. E-mail: a 38-kg load to
mdonelan@sfu.ca generate as
much as 7.4 W
of electricity
during fast
walking. This
device has a
markedly low
“cost of
harvesting”
A The feature of how
me backpack’s humans walk
tab
oli device may provide
c efficiency is another means
po about 31% (5), of economical
we and muscle’s energy
r
COH = ( 1) peak harvesting.
A Muscles
electri efficiency is
cal about 25% (3), cyclically
power
yielding an perform
where A refers expected COH positive and
to the difference of 12.9. But negative
between walk- the backpack’s mechanical
ing while actual COH of work within
harvesting 4.8 ± 3.0 each stride
energy and (mean ± SD) (Fig. 1A) (8).
walking while is less than Mechanical
carrying the 40% of the work is
device but expected required to
without amount. Its redirect the
harvesting economy body’s center
energy. The appears to of mass
COH for arise from between steps
conventional reducing the (9, 10) and
power energy simply to move
generation is expenditure of the legs back
simply related walking with and forth (11,
to the efficiency loads (6, 7). 12). Even
with which (i) No device has though the
the device yet average
converts approached mechanical
the power work
mechanical
performed on
work to elec- generation of
the body over
tricity and (ii) the backpack
an entire stride
muscles convert without the
need to carry a is zero, walking
chemical energy
exacts a
into positive heavy load.
We propose metabolic cost
work
that a key because both

COH for conventional A metabolic power


= generation A electrical power

1 =__________________(2)
device eff x muscle eff

www.sciencemag.org SCIENCE VOL 319 8 FEBRUARY 2008 807


REPORTS
po i ec
sit n el
iv c er
e r ati
an e on
d a (F
ne s ig.
ga i 1 Fig. 1. Theoretical advan-
tiv n B) A Cyclic motion B Cyclic C Cyclic motion tages of generative braking
motion with generative during cyclic motion, com-
e g .
with continuous braking
m t Al paring the back-and-forth
generation
us h th motion of the knee joint
cl e ou G G without power generation (A)
e l gh against a generator operating
w o ge continuously (B) and against
or a ne Limb continually Generator (G) a generator operating only
k d rat accelerates (+) resists acceleration, Generator only during braking (C). Each
assists deceleration assists deceleration
re o in
and decelerates (-) column of plots shows the
more positive rate of work performed by
qu n g
Wor muscles (work rate) and the
ire t el
m h e
k
rate
+ ti me less negative electricity (elect. power)
et e c- - cyclic generated over time, as well
only as the average metabolic
a- m tri power expended by the hu-
Ele
bo u cit ct. man and the resulting average
lic s y ti me
po electrical power (ave. power
en c du wer
bar graphs). In (B) and (C),
er l rin work rate is compared against
gy e g - conven-
COH <tional COH = very low that for (A), denoted by
(3 s th Ave. 0 0 dashed lines, and average
). d e Pow Ave. Ave.
er Pow Pow power is shown as the dif-
C u ac + er er ference (D ave. power) with
very small
ou ri ce respect to (A). COH is
pli n ler defined as the ratio of the
Meta- Me ta - Elec- Met a- Elec-
ng g ati bolic bolic trical bol ic trical electrical to metabolic D ave.
a a on powers.
ge c ph
ne c as
rat e e
or l w
to e ou
le r ld
g a ex
m ti ac
oti o t a
on n su
w b bs
ou u ta
ld t nti
ge a al
ne s m
rat s et
e i ab
el s oli
ec ti c
tri n co
cit g st,
y t do
thr h in Fig. 2. Biomechanical energy harvester. (A) The device has an aluminum chassis (green) and
ou e g generator (blue) mounted on a customized orthopedic knee brace (red), totaling 1.6-kg mass, with one
gh m so worn on each leg. (B) The chassis contains a gear train that converts low velocity and high torque at the
ou du knee into high velocity and low torque for the generator, with a one-way roller clutch that allows for
t d rin selective engagement of the gear train during knee extension only and no engagement during knee
ea u g flexion. (C) The schematic diagram shows how a computer-controlled feedback system determines
ch ri th when to generate power using knee-angle feedback, measured with a potentiometer mounted on the
cy n e input shaft. Generated power is dissipated in resistors. Rg, generator internal resistance; RL, output load
cl g de resistance; E(t), generated voltage.
e, d ce

808 8 FEBRUARY 2008 VOL 319 SCIENCE www.sciencemag.org


ler r po
ati a w
on t er
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ou l ce
ld y ler
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er e H
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et k ou
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(1 v 6).
5).e Th
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so o th
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m t ist
oti s or
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thr h us
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gh g to
a e ch
on n ar
e- e ge
w r a
ay a ba
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ch r Th
, ( e
tra 1 de
n 7 vi
s- ). ce
mi F eff
tti o ici
ng r en
on c cy,
ly o de
kn n fin
ee v ed
ex e as
te n th
ns i e
io e rat
n n io
m t of
oti t th
on e e
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sp ti ctr
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est o Fo
im a r
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C m rat
O p iv
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of r th
6. e e
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(E a rv
q. y est
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to e gy
en k w
ga n he
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on e ve
ly . r
du W th
rin e e
g c kn
th o ee
e m is
en p ex
d a te
of r nd
th e in
e d g
sw t (1
in h 8).
g i W
ph s e
as m co
e o ul
(F d d
ig. e als
3), a o
pr g m
od a an
uc i ua
in n lly
g s dis
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tri c ge
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in e m
de s kn
ce t ee
ler s m
at- e oti
in n on
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th r Th
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ng r w
ag g hil
ed y e
m - w
od h al
e a ki
se r ng
rv v on
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im a e
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th e ate
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m a m
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of d ng
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ha a pir
rv l o
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er s etr
m u y
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en w ea
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of o re
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co r e
st e ele
of a ctr
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in i w
g c er
el e ou
ec o tp
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c e
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rat n lic
or e co
(Fi r st
g. a ov
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C) e th
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In 7 of
th . th
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rat f n
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n l 9).
m e In
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(Fi i ge
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A) y iv
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re e Th
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at i de
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REPORTS
on g es
str e s
at n th
e e an
th r 1
at a W
su ti of
bs v m
ta e eta
nti b bo
al r lic
el a po
ec k we
tri i r
cit n wa
y g s
co ( re-
ul F qu
d i ire
be g d
ge . to
n- 4 ge
er ). ne
at A rat
ed v e1
wi e W
th r of
mi a ele
ni g ctr
m e ici
swing stance al C ty.
0 in O Th
Knee end
mechani -20 A swing cr H is
cal phase harvester
power resists Normal ea i is
(W) harvester knee knee joint se n si
assists positive power
knee work in g g-
Harveste 20 B negative us e nif
r work
mechani er n ica
cal 0 eff e ntl
power
20C ort r y
disengage Continuous generation
Generato generative Generative braking . a les
r braking
electrical T ti s
power 0 engage he v th
generative
braking co e an
knee ext rre b th
0 20 40 60 80 100 sp r e
(red line) and generative % stride cycle on a C
braking (blue line). (C) di k O
Generated electrical power over time, also for both types of generation. ng i H
C n of
O g 6.
A Continuous B C Cost of Harvesting w 4
generation Generative H
50
braking va a ex
7 conventional gen. s pe
lu
∆ 40 CO
6
es o cte
Aver H 5 hi n d
(Di
age 30
Pow me 4 ** gh l for
er nsi
onl 3
lig y co
(W) 20
ess ht 0 nv
) 2 th . en
10 7 tio
1 e
0 0 ad ± na
MetE l e c -
a-t r i c a l
M e t a - Elec-
b o l i c trical
C o n t . Gen. va 4 l
g e n . braking . ge
nt
indicate significant differences with conventional generation (*) and between continuous ag 4 ne
generation and generative braking (**) (P < 0.05 for all comparisons). e ; rat
of l io
n m er
(P e en
= r ce
0. m be
01 o tw
). d ee
Th e n
e a th
C l e
O s tw
H o o
in g m
co e od
nti n es,
nu e 2.
ou r 2
s a ±
ge t 0.
ne e 7
rat d W
io s of
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2. m ctr
3 e ici
± o ty,
3. f ca
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y y eta
lo f bo
we r lic
r o co
th m st
an t of
th h 13
at e ±
for d 12
co e W
nv c (P
en e =
tio l 0.
na e 05
l r ).
ge a A
ne ti C
rat o O
io n H
n o ta
(P f ke
= t n
0. h fro
01 e m
), k th
in n e
di e av
cat e er
in . ag
g T e
th h rat
at e io
th d of
e if th
for f es
e a hi
dif t gh
fer t C
en h O
ce e H
s k an
yi n d
el e pa
ds e r-
5. d tia
7 u lly
± ri by
6. n ge
2, g ne
w w rat
hi a iv
ch l e
is k br
ne i ak
arl n in
y g g
th p wi
e r th
sa o a
m d ve
e u ry
as c lo
th e w
at s C
ex e O
pe l H.
cte e Bu
d c t
of tr ge
co i ne
nv c rat
en it iv
tio y e
na p br
l a ak
ge rt in
ne i g,
rat a wi
io ll th
n y les
(P b s
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ag Bo Huang,1,2 Wenqin Wang,3 Mark Bates,4 Xiaowei Zhuang1,2,3*
in
g Recent advances in far-field fluorescence microscopy have led to substantial improvements in
of image resolution, achieving a near-molecular resolution of 20 to 30 nanometers in the two
bi lateral dimensions. Three-dimensional (3D) nanoscale-resolution imaging, however, remains a
ol challenge. We demonstrated 3D stochastic optical reconstruction microscopy (STORM) by
og using optical astigmatism to determine both axial and lateral positions of individual
ica fluorophores with nanometer accuracy. Iterative, stochastic activation of photoswitchable
l probes enables high-precision 3D localization of each probe, and thus the construction of a 3D
sp image, without scanning the sample. Using this approach, we achieved an image resolution of
eci 20 to 30 nanometers in the lateral dimensions and 50 to 60 nanometers in the axial dimension.
m This development allowed us to resolve the 3D morphology of nanoscopic cellular structures.
en
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