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Study of functional viability of SU-8-based microneedles for neural applications

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2009 J. Micromech. Microeng. 19 025007

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IOP PUBLISHING JOURNAL OF MICROMECHANICS AND MICROENGINEERING
J. Micromech. Microeng. 19 (2009) 025007 (8pp) doi:10.1088/0960-1317/19/2/025007

Study of functional viability of SU-8-based

microneedles for neural applications
Luis J Fernández1, Ane Altuna1, Maria Tijero1, Gemma Gabriel2,3,
Rosa Villa2,3, Manuel J Rodrı́guez4, Montse Batlle4, Roman Vilares1,
Javier Berganzo1 and F J Blanco1
1
MEMS/MST Department, Ikerlan S.Coop, Mondragón, Spain
2
Instituto de Microelectrónica de Barcelona (IMB-CNM), CSIC, Campus UAB, 08193 Bellaterra,
Barcelona, Spain
3
Centro Investigación Biomédica en Red (CIBER-BBN), Spain
4
Unitat de Bioquı́mica, Facultat de Medicina, Institut d’Investigacions Biomèdiques August Pi i Sunyer
(IDIBAPS), Universitat de Barcelona, Barcelona, Spain
E-mail: ljfernandez@ikerlan.es

Received 30 July 2008, in final form 12 November 2008

Published 14 January 2009
Online at stacks.iop.org/JMM/19/025007

Abstract
This paper presents the design, fabrication, packaging and first test results of SU-8-based
microneedles for neural applications. By the use of photolithography, sputtering and bonding
techniques, polymer needles with integrated microchannels and electrodes have been
successfully fabricated. The use of photolithography for the patterning of the fluidic channel
integrated in the needle allows the design of multiple outlet ports at the needle tip, minimizing
the possibility of being blocked by the tissue. Furthermore, the flexibility of the polymer
reduces the risk of fracture and tissue damage once the needle is inserted, while it is still rigid
enough to allow a perfect insertion into the neural tissue. Fluidic and electric characterization
of the microneedles has shown their viability for drug delivery and monitoring in neural
applications. First drug delivery tests in ex vivo tissue demonstrated the functional viability of
the needle to deliver drugs to precise points. Furthermore, in vivo experiments have
demonstrated lower associated damages during insertion than those by stereotaxic standard
needles.

1. Introduction chemotherapy, since a high dose of drugs can be delivered only

Conventional injection of drugs for the treatment of brain
diseases is particularly difficult due to the blood brain
barrier (BBB) [1]. Generally, only low-molecular-mass
lipid-soluble molecules and a few peptides and nutrients
can cross this barrier to a significant extent, either by
passive diffusion or using specific transport mechanisms.
Convection-enhanced drug delivery (CED) is a promising
therapeutic method for treating brain diseases, such as tumors,
neurodegenerative diseases (e.g. Alzheimer), Parkinson and
epilepsy, by enhancing the penetration of drugs [2]. Because
CED uses direct infusion of a drug-containing liquid into
tissue, the transport is dominated by convection. Thus, CED
has the potential of increasing the drug penetration distance
and mitigating the decay in concentration with distance due to
the diffusion process. Also, CED is a way to deliver only local
to the damaged tissue avoiding the problems associated with
chemotherapy in the rest of the human organs.
CED has been tested extensively in humans to treat
different types of brain diseases [3–5]. The results of different
studies and clinical trials indicate that convection can be used
to distribute molecules, regardless of their size or molecular
weight, throughout most regions of the brain. In some clinical
trials, an additional electrode is inserted in order to stimulate
the region where the drug is to be delivered. This cannula is
most often a stainless steel needle ranging from 20 to 32 gauge
in size. In the previous studies, flow rates ranged from 0.1 to
10 μl min−1 and were controlled with an external syringe
pump. At flow rates greater than 1 μl min−1, the backflow
of infused solutions up the outside of the needle shaft has
been reported [6]. Apparently, at sufficiently high flow rates,
the tissue separates from the needle, and the fluid flows

0960-1317/09/025007+08$30.00 1 © 2009 IOP Publishing Ltd Printed in the UK

J. Micromech. Microeng. 19 (2009) 025007 L J Fernández et al

preferentially along the separation. The backflow reduces

the control over drug delivery because infused solutions can
flow out of the brain or into highly permeable white matter,
which tracts surrounding the infusion site. The separation,
which allows the backflow, can be controlled by adjusting
the flow rate and the size of the needle [7]. Also, standard
needles and catheters can induce damage and uncontrolled
drug delivery due to their rigidity. Guarnieri et al report
the replacement of rigid infusion tubes with flexible tubing
increasing the reliability of local CED [8].
Another problem encountered with needles is an
unexpectedly large pressure at the needle tip at the beginning
of an infusion. The large pressure may indicate that the tip
of the needle is partially or fully occluded when it is inserted
into the brain. It has been suggested that a needle or catheter
with several fluid outlets along its side, rather than at its tip,
could alleviate some of these problems [9]. However, standard
cannulas or needles have only got an outlet at the tip.
Silicon micromachining-based microfluidic devices have
been developed during the last 10 years to fulfill the
requirements for CED [10–21]. However, it has been
demonstrated that the rigidity of the silicon devices induces
tissue damage during the insertion, and especially during the
operation of the device [22, 23]. The replacement of silicon Figure 1. Schematic drawing of an SU-8-based microneedle.
by flexible polymers could reduce tissue damage because they
can adapt to the tissues and deform their shapes as the organs of the needle which will be inserted in the tissue has been
deform [22–26]. Moreover, the flexible electrodes may avoid designed to be 3 mm long in order to reach the neural zones
both gradual shift in the recording location and a reduction of interest, while the width is chosen to be 400 μm with
in the signal to noise ratio; these problems often occur a thickness of 220 μm. Although silicon-based microneedles
with rigid electrodes during long-term experiments because can be comparably smaller [10], with typical dimensions in the
of the continued differential motion in the tissue [27]. In order of 100 μm wide and 20 μm thick, the needles presented
addition, most of the polymer materials have a higher degree in this work were designed to ensure their mechanical stability.
of biocompatibility than silicon, making them more suitable to Therefore, the SU-8 needle design presented in the paper is not
be used for chronic implant applications [28]. First polymer- meant to fabricate neural probes directly competitive to silicon-
based microfluidic needles have been fabricated mostly in based neural probes, but as a first step for the development of
polyimide and BCB [29, 30]. However, these materials present polymer-based probes which could improve in the future the
problems related to its excessive flexibility, since they cannot nowadays state of the art. The insertion of the needle to a depth
penetrate into the tissue in a proper way. Furthermore, flow of up to 3 mm allows the infusion of fluids to rodent superior
control elements for chronic implant applications are not yet neuronal areas, such as cerebral cortex, in animal models of
integrated. Therefore, the development of robust microfluidic experimental neuroscience. The microfluidic channel inserted
devices with the rigidity required to penetrate the tissue, and in the microneedle is 50 μm wide and 16 mm long. In order
a certain grade of flexibility to deform their shapes as the to reduce blocking effects during the needle insertion, five
organs deform, is still a challenge. In this work, we propose different outlet ports at the needle tip have been implemented.
the fabrication of microfluidic devices made only of an SU-8 Finally, six different electrodes with a size of 50 × 50 μm2
polymer with integrated metal electrodes. Such electrodes are are placed close to the needle tip (distances between electrodes
not only meant for neural recording but can also be used as are shown in figure 3(b). Such electrodes are meant to be used
a monitoring tool of fluid dispensing. The SU-8 polymer has not only as sensors for neuron activity but also to monitor the
been chosen because of its excellent electrical (high isolation, neuron response to a specific drug delivered by the needle.
low loss), mechanical (low Young modulus and low stress) Fabrication of the needles starts with a temporary bonding
and fluidic properties, and its biocompatibility. of a thin kapton film (125 μm) on top of a Pyrex substrate
(figure 2(a)) [31]. Kapton is used because of its low adhesion
2. Design and fabrication to SU-8, allowing the release of the devices from the substrate
when their fabrication finishes. At the same time, most of
The design of the microneedles is presented in figure 1. Three deposition and etching processes in microfabrication are still
different needle zones can be observed. On the widest section, possible without delaminating problems. Once the kapton
a 5 × 5 mm2 area allows the proper electric and fluidic film is fixed to the substrate, a 160 μm thick SU-8 structural
encapsulation of the needle. Then, an 8 mm long path is layer is deposited on top of it (figure 2(b)). This layer is
needed to progressively reduce the needle size. The part spun on the wafer in two steps, 80 μm thick each. After every

2
J. Micromech. Microeng. 19 (2009) 025007
Figure 2. Fabrication process flow for SU-8-based microneedles.
spinning step, a soft-bake (SB) treatment is performed, heating
the wafer up to 65 ◦ C for 10 min and 95 ◦ C for 30 min. Once
the SB treatment of the last layer has finished, an exposure of
400 mJ cm−2 is performed using a mask which will define
the shape of the microneedle. Next, 50 nm of chromium and
150 nm of gold are deposited by sputtering on top of the SU-8
keeping the wafer temperature below 50 ◦ C (figure 2(c)). In
this way, cross-linking of the un-exposed areas is prevented.
The patterning of the metal layer is performed using
standard photolithography steps and wet chemical etching. As
a result, the un-exposed SU-8 is protected by the chromium
and gold layers from being cross-linked during the required
subsequent processing steps (see figure 2(d)). As a next step,
50 nm of titanium and 150 nm of platinum are deposited and
patterned by a lift-off technique (see figures 2(e)–(g)). In this
way, metallic electrodes could be implemented on top of a
patterned 180 mm thick structure. Then, the 20 μm thick
SU-8 layer which will define the fluidic channel and cover
the metallic traces of the electrodes is spun and patterned
by standard photolithography (figure 2(h)). At this state,
chromium and gold are etched, and all un-exposed SU-8 is
developed from the wafer (figure 2(i)). Using another Pyrex
wafer with a kapton laminated on top, a new 20 μm thick layer
of SU-8 is processed (figure 2(j)) in order to define the cover
of the microchannels present in the needle. This layer includes
the holes to allow the electrical contact to electrodes and the
fluidic connection to the microchannel. Once both SU-8 layers
are generated by photolithographic processes into two different
substrates covered with a kapton film, they need to be bonded
3
L J Fernández et al
together to define a microchannel structure. First, the wafers
are properly aligned and transferred to the bonding chamber.
Misalignment between both wafers previous to bonding was
found to be in the order of 5 μm or less, since both wafers
are transparent. Therefore, a proper alignment with an error
much smaller than any of the needle dimensions is always
achievable. Then, the chamber is evacuated to 10−3 mbar to
remove air gaps between both wafers. Afterward, the wafers
were brought into contact, and temperature and pressure are
increased to 100 ◦ C and 3 bar, respectively, for 30 min [31–33].
Since the cover layer has been processed by photolithography,
inlet and outlet ports are already defined. Finally, devices are
manually released from the kapton film (figure 2(k)). Once
the fabrication process was optimized, the yield was found to
be close to 90%.
As a consequence of the fabrication, electrodes are
recessed 40 μm due to the presence of two 20 μm thick layers
which define the channel and the cover. As a consequence,
signal from neurons might be decreased due the extra distance
between the electrode and the neural tissue.
A picture of a complete microneedle and a close view of
a needle tip from a typical fabrication result can be observed
in figures 3(a) and (b), respectively.
3. Packaging
In order to perform the characterization of the microneedles
in the agarose gel and obtain preliminary data from in vitro
and short in vivo tests, a special package was designed
and fabricated. Figure 4(a) shows a schematic overview
of the packaging developed for the housing of the needles.
The fluidic connection between the outside world and the
microneedle is performed by a channel present in the top
capsule. The use of cylindrical-shaped gaskets guaranties
a perfect sealing without leakage for pressures up to 4 bar.
Finally, the 50 μm wide channel present in the microneedle
allows the fluidic delivery at the needle tip. The electric
connection is made using spring contact pins installed in the
top capsule. In order to prevent any undesired displacement,
the bottom capsule includes a pattern to fit the microneedle
base. Once the needle has been placed on the bottom
packaging piece, two fixed slides are used to align both
capsules, and four screws ensure a proper contact. In this
way, electric and fluidic connections and mechanical fixation
of the needle are automatically performed once the four screws
have been fixed (see figure 4(b)).
4. Characterization
The characterization of SU-8 needles included three different
aspects.
4.1. Fluid delivery characterization
The flow rate as a function of applied pressure was measured
for the SU-8 microneedles when they were immersed in
two different environments, open air and agarose gel, 0.6%
weight/volume (w/v). The agarose gel was selected since
J. Micromech. Microeng. 19 (2009) 025007 L J Fernández et al

(a)

(b)

(a)

(c)

(b)

Figure 4. Microneedle packaging. (a) Package drawing scheme and

(b) the picture of an encapsulated microneedle.

Figure 3. Fabrication results. (a) Top view picture of a finished

microneedle next to a 5 Euro cent coin, (b) the close-up picture of a
microneedle tip showing the electrode configuration and (c) the
SEM picture of a needle tip showing outlet ports.

it mimics the pressure-driven flow in the brain tissue [4].

The SU-8 microneedles showed enough rigidity to penetrate
the agarose gel without observing any deformation, and no
rupture occurred after hundreds of trials. The fluidic setup for
the characterization of the needles started with an EFD 1400
fluid dispensing system (http://www.efd-inc.com/), which was
used to apply controlled pressure on the microneedle. Such a
system was directly connected to an ASL1600-10 mass flow Figure 5. Fluidic characterization of a microneedle in open air and
inserted in agarose (0.6% w/v).
sensor from Sensirion (http://www.sensirion.com/) in order
to measure the flow. Finally, the fluidic circuit ends with its
connection to the microneedle using the packaging described 4.2. Impedance monitoring while delivering in the
in section 3. As can be observed from figure 5, a linear relation agarose gel
between flow rate and pressure was obtained. As expected,
higher fluidic resistance was observed when the needle was In order to validate the electrodes placed at the needle tip,
inserted in agarose. impedance measurements between two of them (the ones

4
J. Micromech. Microeng. 19 (2009) 025007
Figure 6. Monitoring of an SU-8-based microneedle by means of
impedance measurements: t = 36 microneedle is inserted in the
agarose gel (0.6% w/v), t = 79 s saline solution delivery starts with
a flow rate of 2 μl min−1.
placed closest to the tip) were performed using an HP4284A
LCR meter while delivering a saline solution (0.9% of NaCl)
in the agarose gel (0.6% w/v). The electrodes used had an
area of 50 × 50 μm2, and were separated by 250 μm from
each other while the shortest distance to the outlet ports was
100 μm. Impedance measurement results obtained at 1 MHz
are presented in figure 6. As can be observed, a very stable
impedance value is obtained when the microneedle is in open
air (0–36 s).
When the microneedle was introduced in the agarose gel,
a sudden small decrease in impedance was observed (36 s).
The impedance value between the electrodes was also found
very stable when immersed in the agarose gel (36–75 s).
After applying a 2 μl min−1 flow of a saline solution to the
microneedle (75 s), a slow decay in impedance was observed
(75–129 s) until a new stable value was reached (129 s). In
this way, the absorption of the saline solution by the agarose
gel was measured.
4.3. Ex vivo test
A post-mortem experiment with a rat brain was done in
order to analyze the needle’s mechanical and microfluidic
viability. Adult male Wistar rats (body weight 300–350 g)
were obtained from the animal housing facilities of the School
of Medicine (Universitat de Barcelona). They were kept on a
12 h/12 h day and night cycle, and housed with free access to
food and water. Animals were manipulated according to the
European legislation (86/609/EEC). All the efforts were made
to minimize the number of animals used, and their suffering
and procedures were approved by the Ethic Committe of the
Universitat de Barcelona, under supervision of the Generalitat
de Catalunya.
The rats were anesthetized and decapitated. The brains
were quickly removed and placed in an ice-cold glass dish to
perform the experiment. Even though it is flexible, the SU-8-
based microneedle was easily inserted into the parietal cortex
in a perpendicular plane.
To test the device as a drug delivery system, a 0.5%
(w/v) solution of methylene blue in distilled water was used,
5
L J Fernández et al
Figure 7. Flow and total volume of methylene blue delivered into
the rat’s brain by the microfluidic needle.
which was injected at a fluid rate of about 0.5 μl min−1. The
solution was inserted by a microneedle installed in a sterotaxic
instrument. A standard microinjection protocol was followed,
i.e. after completion of the infusion, the needle was left in
place for an additional 3 min to allow for passive diffusion and
to prevent spread of the excitotoxin up the needle track upon
removal; then, the needle was slowly retracted. As can be
observed in figure 7, the solution was injected progressively
without any relevant flow peaks. At the end of the insertion,
a negative flow is observed (around 90 s in figure 7), meaning
that some of the solution was retracted from the tissue through
the microneedle. This was probably due to an excess of
flow rate applied, which could not be completely absorbed.
More experiments will be carried out in the future in order to
minimize this undesired effect.
The brains were frozen after the delivery with powdered
dry ice for further histological analysis. 20 μm coronal slices
were obtained by microtomy at the level of the injection,
in order to analyze the needle placement and the blue of
methylene dispersion in the brain. This analysis showed
that needle penetration depth was up to 3 mm, which means
that the fluid solution was delivered to the V and VI layers
of the cerebral cortex. The analysis also showed good
needle performances with a dispersion of methylene blue in
the internal layers of cerebral cortex and corpus callosum
disruption (see figure 8).
4.4. In vivo test
During sterotaxic procedures, a small relative movement
between the brain and the skull is possible and may result
in small undetectable brain damage. Also an accidental
movement of the needle during the procedure cannot be
discarded. In this line, we tested whether the flexibility of
an SU-8 needle constitutes an advantage versus that of other
needles made from any rigid material. We evaluated the
mechanical damage produced by the SU-8 needle in a rat’s
brain during its insertion, and a comparison with a stereotaxic
standard needle was performed.
J. Micromech. Microeng. 19 (2009) 025007
Figure 8. Coronal section of the rat’s brain evidenced the location
of the injection. Blue staining in the prefrontal cortex clearly shows
the methylene blue injection and diffusion with a corpus callosum
disruption (arrow).
Under equithesin anesthesia (a mixture of chloral hydrate
and sodium pentobarbital; 0.3 ml/100 g body weight,
intraperitoneally), rats were placed in a stereotaxic instrument
(David Kopf, Carnegie Medicin, Sweden) with the incisor
bar set at −3.3 mm. After opening of the head skin with
a scalpel, the skull was drilled and the dura mater broken.
Needles were then introduced in the rat’s dorsal hippocampus.
The stereotaxic coordinates were 3.3 mm caudal to bregma,
2.2 mm lateral to bregma and 2.9 mm ventral from dura
mater, according to the Atlas of Paxinos and Watson [34].
The SU-8 microneedle was inserted into the right hemisphere,
and stereotaxic standard needle (0.28 mm external diameter)
was inserted into the left one. The needles were moved
±1 mm caudal from the initial point (in the perpendicular
plane with respect to the needle direction), and then slowly
retracted. Twenty-four hours later, the rats were anesthetized
and sacrificed. The brains were removed, frozen with
powdered dry ice and stored at −80 ◦ C.
Adjacent coronal 14 μm sections from the whole
hippocampus were obtained from all brains and mounted on
slices. Standard Nissl staining was performed to evaluate
the neuronal loss and the morphometry of cerebral cortex
and hippocampus. Microglial cells were identified by
histochemical procedures using Isolectin B4 (IB4), a Griffonia
Simplicifolia extract that binds to microglial galactose-
containing glycoconjugates. IB4 was used conjugated with
biotin, and detected by the biotin-avidin-peroxidase method.
Briefly, sections were incubated overnight at 4 ◦ C with IB4
(Sigma, St Louis, MO) diluted 1:25 in normal goat serum, that
is, 1:100 volume/volume (v/v) in 0.01 M PBS; pH 7.4. After
incubation with ExtrAvidin (1:250; Sigma, St Louis, MO),
6
L J Fernández et al
(a) (b)
(c) (d)
Figure 9. Cresyl-violet staining of the brain sections: (a) stereotaxic
standard needle injection and (b) SU-8 needle. IB4 histochemistry
showed the area of microglial reaction associated with the lesion:
(c) standard needle injection and (d) SU-8 needle. The areas of
lesion and microglial reaction are delimited by arrow heads.
sections were developed in a 0.05 M Tris solution containing
0.03% (w/v) diaminobenzidine (DAB) and 0.006% v/v H2O2.
Nissl staining was used to assess neuronal injury in terms
of area of neuronal loss evidenced by a lack of staining
(figures 9(a) and (b)). No significant neuronal loss was
detected in the dorsal hippocampus after the movement of
the needles. Still, this mechanical movement showed a small
area of cellular loss in the case of SU-8 needle insertion
evidenced in the cerebral cortex as a small area of reduction
of staining (figure 9(b)). However, this area of decreased
staining was much higher for the case of the insertion of
the standard needle (figure 9(a)). IB4 histochemistry was
performed in order to stain the area of inflammation mediated
by a microglial reaction, which is evidenced as an increasing
of staining. IB4 histochemistry evidenced a higher area of
inflammation associated with microglial reactivity in response
to the standard needle movement (figure 9(c)), when compared
with the SU-8 one (figure 9(d)). Thus, we can conclude that
due to its flexibility, the tissular injury caused by the SU-
8 needle is lower than the one caused by standard needles
currently used in stereotaxia procedures.
5. Conclusions
The work presented demonstrates the functional viability of
SU-8-based microfluidic needles. This device, with integrated
microfluidics and electrodes, can be used in neurophysiologic
studies to deliver drugs to precise points, which is a very
important feature especially concerning in vivo studies of
brain functions. Following these good results, the device will
now be adapted to a standard stereotaxic equipment. Thus,
the system will be suitable to be used on in vivo short-term
J. Micromech. Microeng. 19 (2009) 025007
experiments through acute administration of compounds in
specific brain nuclei, as well as in long-term approaches with
a chronic administration of compounds through a permanently
implanted needle. Although the viability of SU-8-based
microneedles has been demonstrated in this work, some
improvements are needed in order to be competitive to those
based on silicon technology. The authors are now busy with
a new design in order to make them thinner and narrower,
which will require adjustments in the fabrication process.
Furthermore, a smaller packaging for chronic testing of the
microneedles is required, and will be developed in the near
future.
Acknowledgments
The authors thank Dr N Mahy (Unitat de Bioquı́mica,
Facultat de Medicina, IDIBAPS, Universitat de Barcelona) for
stereotaxic technical support and biomedical scientific advice.
M Batlle was the recipient of a fellowship from IDIBAPS.
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