AbstractThe mosquito proboscis is composed of seven

micro parts. Among them, labrum and maxillae play important

roles in insertion. The labrum is a central needle which sucks
blood; two maxillae are left/right needles aside the labrum,
respectively. They cooperatively move, i.e., one maxilla goes
forward, labrum goes forward, another maxilla goes forward,
successively. We noticed that the maxilla has a jagged
harpoon-like shape on its tip area, which has an anchoring effect.
We proposed the easy insertion mechanism as follows: the
equivalent stiffness, which is the ratio of the resistance force
relative to the moving distance, is considered. The total stiffness
in pushing out the central needle can be made smaller than zero
due to the anchoring effect of the side needles. We named it as
negative stiffness mechanism. Equivalent stiffness during
pulling/pushing a fabricated needle was experimentally
estimated, which indicated the validity of proposed mechanism.
I. INTRODUCTION
low-invasive needle is strongly desired in many medical
treatments such as biopsy, transdermal drug delivery,
neural interface, lancets for puncturing and bleeding diabetics.
In particular, diabetics have to collect their blood for the
glucose level measurement, which is indispensable for health
monitoring. Their skin is punctured by a lancet, which is a
solid metal needle having straight shape, causing small
bleeding. This operation is painful and fearful. On the other
hand, a human being has almost no pain and no fear when pricked
by a needle of mosquito, i.e., the mosquito proboscis should be
a good model for a painless insertion tool. One of the reasons
for painless insertion of mosquito proboscis is considered to
be its thinness, the diameter of which is approximately 50 m
[1, 2]. Pain spots are located at intervals of about 1 mm on a
human skin [3]. As the needle becomes thin, its possibility to
evade the spots becomes high. Based on this consideration,
many researches for achieving painless needles have been
conducted, in which main concern is to make the needle
thinner than the present commercial needle having diameter
of approximately 300 m [4-9]. Commercial lancet needles
are also thinned and distributed recently. For example, a

S. Aoyagi, Y. Takaoki, C. Huang, T. Tanaka, M. Suzuki, and T.
Takahashi are with Department of Mechanical Engineering, Kansai
University, Suita, OSAKA 564-8680, JAPAN
T. Kanzaki is with Dainihon Jochugiku Co., Ltd., Toyonaka, OSAKA
561-0827, JAPAN
T. Matsumoto is with Graduate School of Medicine, Dentistry and
Pharmaceutical Sciences, Okayama University, Okayama, OKAYAMA
700-8530, JAPAN
(Corresponding author: Seiji Aoyagi, phone: +81-6-6368-0823; fax:
+81-6-6388-8785; e-mail: aoyagi@kansai-u.ac.jp).

needle of 200 m in diameter is distributed by Terumo Corp.,
which is fabricated using press working of metal [10]. These
reported needles relatively lessen the pain compared to the
currently used needle; however, a completely painless needle
has not been invented yet by the thinning method. Moreover,
thinning the needle causes another problem of its easy
buckling. There apparently exists the limit for reducing the
pain in the way of thinning a sole needle and simply pushing
it against the skin.
We previously observed the mosquito proboscis by a
scanning electron microscope (SEM) [11, 12]. It is composed
of several parts, which are labium, labrum, pharynx, two
maxillae, and two mandibles, as shown in Fig. 1. The labrum,
pharynx, maxillae, and mandibles are bundled and
overlapped by the sheath-like labium. Among them, labrum
and maxillae play important roles in insertion. The labrum is a
central needle which sucks blood; two maxillae are left and
right needles aside the labrum, respectively. We also
previously observed the cooperative inserting motion of
mosquito proboscis using a high-speed camera system. It was
proven that the central and the side needles are advanced
alternatively, the vibration frequency of which is at several
Hz, while the total three needles are gradually moved forward
[12, 13]. Considering these results, the key to elucidate the
mosquitos painless insertion is not only the thinness of
Equivalent Negative Stiffness Mechanism Using Three Bundled
Needles Inspired by Mosquito for Achieving Easy Insertion
Seiji Aoyagi, Member, IEEE, Yutaka Takaoki, Hiroki Takayanagi, Chih-hao Huang, Takahiro Tanaka,
Masato Suzuki, IEEE, Tomokazu Takahashi, IEEE, Tsutomu Kanzaki, Takuya Matsumoto

A
Fig. 1. Magnified images of mosquito proboscis. (a) Optical
image of overview. (b) Schematic composition of seven parts.
(c) SEM image of labrum and maxilla. (d) Magnified SEM
image of tip of maxilla. Harpoon-like jagged shape is seen.
(a)
Labium
Head
Bundled 6 parts of two
maxillae, labrum, two
mandibles, pharynx
Pharynx
Maxilla
Labrum
Labium
Mandible
(b)
(c)
Maxilla

Labrum

1000 10 m
(d)
Maxilla

Jagged shape
850 20 m

2012 IEEE/RSJ International Conference on
Intelligent Robots and Systems
October 7-12, 2012. Vilamoura, Algarve, Portugal
978-1-4673-1736-8/12/S31.00 2012 IEEE 2295



proboscis but also its composition of several parts and their
skillfully cooperative motions. There are only few reports to
pay attention to the way of insertion about commercial metal
needles [14, 15], and no reports about microneedles besides
our previous ones [12, 13]. In the course of observation, we
also found that the mosquito moves not only proboscis but
also its head during insertion. Therefore, the observation of
the head part besides the proboscis part is important for
analyzing the insertion mechanism.
In this paper we noticed that the maxilla has a jagged
harpoon-like shape on its tip area (see Fig. 1), which may
have an anchoring effect when the labrum is inserted forward.
The remainder of this paper is organized as follows: i) we
further observed the inserting motion of mosquito proboscis
to a transparent artificial skin in detail, considering the effect
of jagged shape. ii) Based on the observation, we proposed an
insertion mechanism of mosquito proboscis, in which the
large frictional force between the outer jagged shape of
maxilla and the skin organization plays an important role. The
ratio of the resistance force relative to the moving distance
was considered equivalently to be the stiffness in the moving
mechanism of needle. The stiffness of pulling up the maxilla
is considerably large due to its jagged harpoon-like shape on
tip part. It could make the total stiffness of the inserting
mechanism of labrum even smaller than zero. We named it as
negative stiffness mechanism. iii) Equivalent stiffness
during pulling/pushing a fabricated straight/jagged needle
was experimentally estimated. Using the obtained data, the
validity of proposed mechanism was discussed, considering
not only the motion of needles but also the motion of body
including head, jaw, and legs.
II. OBSERVATION OF MOSQUITOS INSERTING MOTION
A. Setup of Observation System
Several hypotheses of mosquitos insertion mechanism
have been reported by other groups [1, 16]; however, none of
them has been confirmed either theoretically or
experimentally in details. We have tackled this problem using
a high speed camera system, which is equipped with a
magnified lens having a long working distance [12, 13].
Transparent soft slime material, of which Youngs modulus is
approximately 0.2 MPa, is used as an insertion object of
mosquito in the experiment of this article. The slime is
prepared by mixing polyvinyl alcohol (10 g), borax (0.5 g),
and water (15 ml). The reason of using transparent material is
to observe needles motion equivalently underneath the skin.
The experimental system is shown in Fig. 2. Compared
with our previous research, one high-speed camera system
was added, i.e., two camera systems were totally used. It is
because as follows: we plans to simultaneously observe both
the proboscis and the body including head and jaw. However,
at the magnification enough to observe the maxillas jagged
parts of which pitch is several m, one camera cannot observe
the jaw and the head at the same time, as shown in Fig. 3.
Even at the low magnification in this figure, two images
should be superimposed for observing both the proboscis and
the body. Therefore, two parts were exclusively observed by
two different cameras in this article.
Two sets of high-speed camera (NAC Image Technology
Inc., MEMRECAMfx RX-6) were used. One camera is for
obtaining the magnified view of proboscis tip, in which a lens
with high magnification (Leica Microsystems, Z16APO,
optical maximal magnification: 115, working distance: 20
mm) is used. Note that the working distance is long enough to
allow the stereoscopic observation of the mosquito proboscis.
Another camera is for obtaining the overview which includes
head, jaw, and proboscis, in which a lens with moderate
magnification (Tokina Corp., KCM-Z4.5, 4.5, 90 mm) is
used. These two camera systems can capture images
synchronously in time domain.
Considering that mosquitoes gather to the human body by
sensing his temperature of about 36C, the temperature of slime
material was controlled to be this value by using a heater made of
Nichrome (NiCr) wiring. As the mosquito hates visible shiny
lightning, invisible infrared illumination was employed.
Mosquitoes bred up in pure environment were supplied by
Dainihon Jochugiku Co., Ltd. Five mosquitoes were released
in a cage, on the wall of which the slime material was set. A
human operator waited for the mosquito to touch down on the
slime. Once the mosquito begins to insert its proboscis to the
Fig. 3. Captured images for head and proboscis. Two
images are necessary as far as using one camera system.
Labium
(sheath)
7 500 m
Head
Magnification
Labrum
10 100 m
Schematic
Maxilla
7 500 m
Maxillary palp
Proboscis
Fig. 2. Experimental system for observing inserting
motion of mosquito using two high speed cameras.
High speed camera
(1,000 frame/s)
Power source for
heater
For magnified
view for proboscis For overview
including head
Lens
(115, W.D. 20 mm)
Lens
(4.5,
W.D. 90 mm)
Slime sheet equipped with
heater (Youngs modulus
~0.2 MPa)
Inside
Inflared
illumination
Mosquito
Cage
2296



slime, the operator adjusts the direction of lens axis and
controls the focus manually.
B. Observed Results of Inserting Motion
Inserting motion of mosquito proboscis to the transparent
slime material was observed at 1,000 flame/s. Based on the
observation, we found that the mosquito moves its maxillary
palp and maxilla synchronously as follows: it pushes out the
maxilla and raises the maxillary palp at the same time.
Conversely, it pulls up the maxilla and lowers the maxillary
palp at the same time. The left and right maxillary palps are
located on the base of jaw, so the observation of them is easy.
We can grasp the moving direction of maxilla indirectly by
observing the motion of maxillary palp. We also found that
the mosquito moves its body, head, and labrum
synchronously as follows: it pushes out the labrum and lowers
both the body and the head at the same time. Conversely, it
pulls up the labrum and raises the head at the same time, while
it keeps the body at the same height level.
Observation results are shown in Fig. 4, which are as
follows: (Phase 1) one maxilla goes forward while the
corresponding maxillary palp moves upward. At the same
time, the labrum goes backward while the head moves
upward. (Phase 2) The labrum goes forward while the head
and the body move downward. At the same time, two
maxillae go backward while their maxillary palps move
downward. (Phase 3) Another maxilla goes forward. At the
same time, the labrum goes backward. (Phase 4) The labrum
goes forward. At the same time, two maxillae go backward.
This phase is identical to Phase 2. These series of operations
are repeated at several Hz (approximately 2 Hz, ranging from
1 to 7 Hz), while the total three needles gradually moves
forward. At Phases 1 and 3, only the head moves upward and
the body keeps its height, whereas at Phases 2 and 4, not only
the head but also the body move downward. Eventually, the
head and the body gradually move downward, which makes
the three needles gradually move downward. These motions
are schematically shown in Fig. 5.
An example of actual movement of mosquito three needles,
which is obtained using the high speed camera systems, is
shown in Fig. 6.
C. Investigation on Muscle Distribution of Mosquito
Although there are some literatures on dissecting and
observing the mosquito proboscis [17], the muscle
Fig. 6. An example of actual movement of mosquito
three needles based on observation. (a) Labrum. (b) Right
maxilla. (c) Left maxilla.
0
10
20
30
40
50
60
70
80
90
0 0.1 0.2 0.3 0.4 0.5
(a)
(b)
(c)
Downward
Time s
D
i
s
p
l
a
c
e
m
e
n
t

m

Proboscis
Proboscis
Labium
(sheath)
Head
4.5 1 mm
100 m
50
Labrum
Maxilla
Maxillary palp
Phase 1
Phase 2
Phase 3
Phase 4
Movement
of head
Movement
of body
Movement of
maxillary palp
Fig. 4. Observation results.
Soft slime sheet
Up Down
Fig. 5. Schematic motion of mosquito proboscis based
on observation. The head and three needles are
gradually lowered from Phase 1 to Phase 4. These
phases are repeated at several Hz (2Hz, ranging 1-7Hz).
Moving direction
Head
Labrum
Maxilla
Phase 1 Phase 2 Phase 3 Phase 4
2297



distribution related to proboscis motion has not been cleared
yet. We tackled this problem. The dead mosquitoes were
soaked in formaldehyde. After taken out, they were
embedded in paraffin. The bulk paraffin embedding
mosquitoes was sliced to thin films using a microtome (Leica
Microsystems, RM2125RT), each thickness of which is 5 m.
The films were stretched and stuck on preparation glasses,
followed by a staining process using hematoxylin and eosin
solutions. An example of results is shown in Fig. 7. The
stained area corresponds to a cell nucleus. Taking account
that cell nucleuses are lined up in muscle fibers, muscle
distribution can be obtained. Observing many samples, we
have judged that the muscles are only located on the two
places in the jaw, which move two maxillae, respectively. We
have also found that there are no muscles for moving the
labrum, and no muscles on the needles themselves. It means
that the mosquito moves two maxillae by two muscles of the
jaw, and it moves the labrum by its head and body.
III. PROPOSAL OF EQUIVALENTLY NEGATIVE STIFFNESS
MECHANISM
We noticed that the maxilla has a jagged harpoon-like
shape on its tip area. This shape is said to be functional for
easy insertion [1]. There are many hypotheses for the function
of jagged shape [1, 12]: however, none of them has been
confirmed either theoretically or experimentally at the present
state. One of hypotheses is that the jagged parts may have the
function to anchor the maxillae to the skin organization and
may assist the labrums easy insertion by some mechanism,
which is focused on in this section.
Based on the observation results mentioned in the previous
section, an analytical model of mosquitos head and proboscis
including muscles is constructed, as shown in Fig. 8. In the
following of this section, this figure is referred to.
As for the maxilla, the following equation holds true:
(1)
where 1 i means that the right maxilla is considered, and
2 i means that the left one is considered. i
m is the mass of
maxilla, i
x is the displacement of maxilla (downward is
positive direction),
i skin
f is the frictional force given by skin
organization,
i muscle
f is the driving force given by the muscle
located at the base of the jaw.
As for the labrum with head, the following equation holds:
(2)
where c
m is the total mass of head and labrum,
c skin
f is the
frictional force given by skin organization,
c muscle
f is the
driving force given by the muscle located between the body
and the head (see Fig. 8), the term of
1 2 )
(
muscle muscle
f f is
the reactive force given by muscles for driving maxillae.
As the insertion displacement becomes large, the frictional
force becomes large due to the increase of interface area
between the needle and the skin organization. Therefore, the
following equations hold true for the maxillae:
(3)
where
i forward
k is the equivalent stiffness when the maxilla
moves forward, i.e., downward, and
i back
k is that when it
moves backward, i.e., upward. Due to the harpoon-like
jagged shape on maxillas tip area,
i forward i back
k k holds true.
The validities of these equations have been experimentally
confirmed using microneedles fabricated by
micro-electro-mechanical systems (MEMS) technology
(details are explained in the next section). Similarly, the
following equation holds true for the labrum with head:
(4)
The mass of maxilla is negligible compared with those of
head and labrum, i.e., holds true. Although the three
needles move synchronously with certain phase differences,
they gradually go forward, which means their displacements
are assumed to be approximately the same level, i.e., i c
x x
holds true.
Under these assumptions, the following equation is
obtained using Eqs. (1)-(4) in the case that the labrum goes
forward and the two maxillae go backward, i.e., in Phases 2
and 4 shown in Figs. 4 and 5:
(5)
In this equation, the term of ( 2 )
c forward i back
k k is the
equivalent stiffness in pushing out the labrum. If this term can
be set below zero, the mechanism would have equivalently
negative stiffness. In other words, the central needle is easily
inserted to the skin by no forward driving force on it ideally,
while backward driving forces are applied on the side needles.
We named this mechanism as negative stiffness
mechanism. Considering that the stiffness of pulling up
maxilla is considerably large due to anchoring effect arising
from its jagged harpoon-like shape, there is possibility that
the mosquito is actually utilizing this mechanism. This
Fig. 7. Stained sectional cell organization of mosquitos
jaw area. Two muscle fibers are seen, which drive two
maxillas. There is not a muscle fiber for driving labrum.
Muscle fiber
m
i
x
i
=f
i skin
+ f
i muscle
, i=1,2,
m
c
x
c
=f
c skin
+f
c muscle
- f
1 muscle
- f
2 muscle
,
f
i skin
= -k
i forward
x
i
(x
i
>0), k
i back
x
i
(x
i
<0) ,
f
c skin
= -k
c forward
x
c
(x
c
>0), k
c back
x
c
(x
c
<0) .
m
c
x
c
+( k
c forward
2k
i back
)x
c
=f
c muscle
.
m
i
m
c
Fig. 8 Analytical model of mosquitos head and proboscis.
(a) Schematic front view. (b) Model including muscles. m
c
Head and labrum, m
1
Right maxilla, m
2
Left maxilla.



Magnified
proboscis
(a)
x
c
x
1
, x
2
m
2

Leg
m
1

Head
Body
m
c
(b)
Muscle
2298



mechanism could be applied to artificial medical needle
systems for reducing the pain in future.
IV. EXPERIMENTAL INVESTIGATION ON EQUIVALENT
STIFFNESS USING ARTIFICIAL SILICON MICRONEEDLE
IMITATING MOSQUITO
A. Purpose of Experiment
It is difficult to estimate the actual parameters of mosquito
used in the model described in the previous section.
Especially, we want to know the parameters of
c forward
k ,
c back
k ,
i forward
k , and
i back
k of mosquito, and we want to
investigate whether the negative stiffness mechanism holds
true or not. However, the estimation of these parameters is
impossible at present state, limited by the available
measurement instrumentation. Therefore, we fabricated
microneedles artificially, which really imitate mosquito
proboscis in terms of shape and size. Mosquito proboscis is
made of organic material of chitin. It is difficult to fabricate a
real imitation using this biomaterial at present. Thus, we
employed silicon material, which has a good compatibility to
MEMS fabrication. Although the mechanical properties such
as Youngs modulus are different from mosquitos, the
experimental data of
c forward
k ,
c back
k ,
i forward
k , and
i back
k
using the silicon imitation would be useful for discussing the
proposed equivalent stiffness issue.
B. Fabrication of Microneedle Using MEMS Technology
Silicon microneedles were fabricated using MEMS
technology. The fabrication process is shown in Fig. 9. A thin
silicon wafer having 50 m in thickness is employed. The
surface of it is oxidized, followed by photolithography to
define the etching part of silicon. Using the defined SiO
2
area
as an etching mask, silicon is etched by deep reactive ion
etching (DRIE) process using SF
6
/C
4
F
8
gas. Photoresist and
SiO
2
are removed by wet etching, leaving a silicon
microneedle, which has two-and-half dimensionally sharp tip.
For sharpening a tip to make a real three dimensionally
conical shape, an electrochemical etching method, which was
invented by the authors [13], were employed, as shown in Fig.
10. Optical microscope images of the fabricated microneedles
are shown in Fig. 11. One is straight shaped; another is
harpoon-like jagged shaped. Almost the same size and the
shape as mosquitos proboscis were successfully achieved.
There sizes are far smaller compared to currently used
commercial stainless needles.
C. Experimental Estimation of Equivalent Stiffness
The resistance force during inserting/extracting the
fabricated needles to/from an artificial skin of silicone rubber
was detected, as shown in Fig. 12 [12]. A silicone rubber
(thickness is 1 mm, Youngs modulus is 2.2 MPa) is attached
to a load cell (the rated load is 2 N, the linearity is 0.5% to the
full range). A personal computer can detect displacement and
load in real time with resolution of 0.5 mN. As for further
details of system, please see ref. [12], due to want of space.


Experimental result of the resistance force relative to the
forward moving distance is shown in Fig. 13. That in the
backward case is shown in Fig. 14.
The equivalent stiffness, which is the ratio of the resistance
force relative to the moving distance, can be obtained using
the data in Figs. 13 and 14. They are summarized in Table I.
D. Discussion
It is confirmed that the equivalent stiffness in the backward
motion of jagged needle takes large value due to anchoring
effect. Assume the fabricated needles are used corresponding
to the mosquitos central labrum and side two maxillae, then,
the term of ( 2 )
c forward i back
k k , which is equivalent stiffness
in pushing out central needle, takes the minus value of -45
gf/mm according to Table I. It indicates that the proposed
Chemical reactions:
Si + 2H
2
O SiO
2
+ 4H
+

SiO
2
+ 6HF H
2
SiF
6
+ 2H
2
O
Fig. 10. Schematic of electrochemical etching for
sharpening a needle.
Hydrofluoric acid (HF) 10%
Anode Cathode
Si Si



Condition:
200 V, 100 mA, 10~15 s
Fig. 9. Micromachining process of silicon needle.
(d) Etching Si using DRIE
(b) Photolithography
(e) Electrochemical etching of
tip of needle after removal
of photoresist and SiO
2

(c) Etching SiO
2
by CHF
3

Si
(a) Oxidation
SiO
2

Photoresist Sharp tip
Length: 2 mm
Width: 30 m
Thickness: 50
m
Length: 2 mm
Width: 15 m
Thickness: 50 m
Enlargement
Fig. 11. Optical images of the fabricated needles with
sharp tip. (a) Straight needle. (b) Jagged needle.
(a) (b)
2299



negative stiffness mechanism holds true in this experimental
case, which would make the insertion process easy.
On the other hand, the equivalent stiffness in pulling out
central needle, which is expressed as ( 2 )
c back i forward
k k , also
takes the minus value of -2 gf/mm. The results indicate that
the mosquitos body supporting its proboscis is always pulled
to the human skin independent of labrum moving direction.
We anticipate that the legs support the body against the
pulling force. In the contrary case that body is pushed against
the skin, the body is lifted up; the legs can no longer support it,
since legs seem not to have anchoring function on their tip.
V. CONCLUSIONS
Based on the observation using two high speed camera
systems, the easy insertion mechanism using three needles
was proposed. It is as follows: the equivalent stiffness, which
is the ratio of the resistance force relative to the moving
distance, was considered. The total stiffness in pushing out
the central needle can be made smaller than zero due to the
anchoring effect of the side needles. We named it as negative
stiffness mechanism. Microneedles were fabricated using
MEMS technology. The obtained experimental equivalent
stiffness showed the validity of proposed mechanism.
In the present paper, the proposed mechanism was
preliminarily discussed using experimental data on a single
needle. The further experimental discussion using three
needles like real mosquito proboscis is the ongoing work.
ACKNOWLEDGMENT
This work was supported by JSPS.KAKENHI(23656188).
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Mechanics of a mosquito bite with applications to microneedle design,
IOP journal on Biomimetics and Bioinspiration, vol. 3, 2008, pp.
0460001.
[17] A. N. Clement, The Biology of Mosquitoes, CABI Publishing, 2000.
Fig. 12. Experimental equipment for detecting force during
insertion and extraction of needle. Details are in ref. [12].
Strain gauge amp.
Encoder
pulse
Silicone rubber
Load cell
Encoder (8000 pulse/R)
Controller
Program
PC
PC
Command
Motor
Voltage
Microneedle
One-axis moving stage
Holder
TABLE I
EXPERIMENTAL RESULTS OF EQUIVALENT STIFFNESS
Straight needle (gf/mm) Jagged needle (gf/mm)
Forward
c forward
k =11
i forward
k =7
Backward
c back
k =12
i back
k =28

0
1
2
3
4
5
6
7
8
0 0.1 0.2 0.3 0.4 0.5
Jagged needle
Straight needle
Fig. 13. Experimental result of resistance force in forward motion.
Straight needle
Jagged needle
c forward
k

i forward
k
R
e
s
i
s
t
a
n
c
e

f
o
r
c
e

[
g
f
]

Displacement [mm]
-6
-5
-4
-3
-2
-1
0
1
0 0.1 0.2 0.3 0.4 0.5 0.6
Jagged needle
Straight needle
Fig. 14. Experimental result of resistance force in backward motion.
Straight needle
Jagged needle
R
e
s
i
s
t
a
n
c
e

f
o
r
c
e

[
g
f
]

c back
k
i back
k
Displacement [mm]
2300