AbstractBy placing axons into polymeric micro-

channels hosting embedded electrodes the extracellular

amplitude of action potentials is greatly increased,
allowing for robust recording and noise suppression. We
are developing such an electrode interface to record
electrical activity from bladder afferents to restore
bladder control in patients suffering from spinal cord
injury. Here we describe our microchannel electrode
interface in terms of design, microfabrication and
electrode characteristics and report on in vivo bladder
function after implantation of teased dorsal rootlets
within microchannels.
I. INTRODUCTION
HE dysfunction of urinary organs are one of the most
important consequences from spinal cord injury (SCI)
[1]. The repeated catheterization leads to chronic urinary
tract infection, reflex voiding occurs unpredictably leading
to social distress, and overfilling can produce autonomic
dysreflexia. Urination in a voluntary context requires neural
information to be sent to and from the brain via the spinal
cord. Information about bladder fullness is carried to the
sacral cord via the sacral dorsal roots. Local reflex control
and descending information from the brain are integrated in
the lumbar and sacral cord to control the bladder.
The most widely used neuroprosthesis for bladder control
in SCI patients is the Brindley system a sacral root
electrical stimulator [2, 3]. Combined with dorsal sacral
rhizotomy, the device has shown good results in
management of neurogenic dysfunction of the bladder [4].
However the rhizotomy of the afferents that innervate the
bladder usually affect bladder sensation and genital
functions.
In this study, we are designing a recording implant to be
used in combination with the Brindley stimulator that will
detect bladder pressure rises from the neural signal at the
nerve roots. One of the challenges is to define how to
reliably detect neural signals of V range amplitude during

Manuscript received January , 2011. This work was supported by the
EPSRC Translational Grant (No. EP/H00727X/1) on Microchannel
electrode neural interfaces: restoring bladder control and the Royal
Society.
S. P. Lacour (corresponding author; phone: +41 21 6931181; e-mail:
stephanie.lacour@epfl.ch) was with the department of Engineering,
University of Cambridge, UK, and is now with the Institute of
Microengineering, EPFL, Lausanne Switzerland.
D. Chew and J. Fawcett are with the Centre for Brain Repair, University
of Cambridge, Cambridge CB20PY UK (email: dc501@cam.ac.uk;
jf108@cam.ac.uk).
E. Delivopoulos is with the department of Engineering, University of
Cambridge, Cambridge CB30FF UK (email: ed369@cam.ac.uk).
bladder filling. Such extracellular signals are below the
noise threshold for even very good amplifiers and are likely
to be swamped by interference from cutaneous receptors
during ordinary activity (e.g. sitting).
We have demonstrated that microchannel electrodes can be
used as efficient axonal amplifier, i.e. the amplitude of the
recorded extracellular signals can be as large as hundreds of
microvolts [5, 6]. When the microchannel cross-section is
small enough, typically less than 100m side, the effective
extracellular impedance within the channel increases and
larger amplitude of the action potential can be recorded [7].
Based on this design, we are developing a compatible
dorsal root recording device to detect and record bladder
afferent activity from the dorsal roots. In this paper, we
present the design of our microchannel prosthesis, report on
bladder function at 28 days post-implantation of 100 m
2

cross-section microchannel device, and present preliminary
neuronal activity recordings in terminally anaesthetized rats.
II. AXONAL AMPLIFIER
Implantable neural electrodes rely on extracellular
recording; the detected signal corresponds to the change in
the outside potential of the axon membrane as an action
potential passes the electrode. Because of the low
impedance of the surrounding extracellular fluid, this
potential change is very small (< 100 V). This may be
overcome by taking the axons into micro-channels of around
100 m x 100 m cross-section made in an electrically
insulating substrate: the micro-channel restricts the
extracellular space, augments the extracellular impedance
thus increases the recordable amplitude of the action
potential. Furthermore confining of the axon into a narrow,
insulating tubular device forces the extracellular currents to
flow purely longitudinally, parallel to the axon and thus any
electrode within the micro-channel can detect the AP
independently of the location of the nodes of Ranvier [5].
Even with the signal amplification provided by the
microchannel confinement, the extracellular signals
generated by APs travelling in peripheral axons are small
(<10mV) and superimposed on substantial background noise
including electrical, biological and intrinsic noises. By
using a tripolar recording configuration in the micro-
channels, the interference can be reduced dramatically [8].
The active electrode is therefore positioned in the middle of
the microchannel with the outer electrodes equidistant from
it and close to the channel endings.
Microchannel electrode interfaces to assess bladder afferent activity
Daniel J. Chew, Evangelos Delivopoulos, James W. Fawcett, Stphanie P. Lacour, IEEE Member
T
978-1-4244-4141-9/11/$25.00 2011 IEEE 249
Proceedings of the 5th International
IEEE EMBS Conference on Neural Engineering
Cancun, Mexico, April 27 - May 1, 2011
ThE1.2

III. MATERIALS AND METHODS
A. Microfabrication
Microchannel electrodes are manufactured using standard
microfabrication process, silicone elastomer and thin gold
film electrodes.
Two types of microchannel implants are prepared. Passive
implants (without electrodes) are prepared by casting of
silicone elastomer (Sylgard 184, Dow Corning) against an
SU8 mold. The latter is prepared by spin-coating a 100 m
thick SU8 2100 film on a clean silicon wafer. After pre-
baking, the negative photoresist is then UV exposed and
developed to define parallel micro-channels. Before PDMS
casting, the molds are perfluorinated to reduce adhesion
between the PDMS and resist layer. Fig. 1 illustrates a
cross-section of a 6 100-m wide and tall PDMS micro-
channel device. A thin PDMS lid sealing each channel
completes the 5mm long open-top channel array.

Figure 1. Optical micrograph of a passive, open-top, PDMS microchannel
implant. The channels are 100 m side.
Active devices are prepared in three steps. A sketch of the
device is presented figure 2. First the electrode tracks are
evaporated and patterned on the elastomeric substrate; then
the recording sites and contact pads are defined in the
encapsulating elastomer; finally SU8 microchannels are
patterned on top of the microelectrode array. Similarly to
the passive implants, a thin PDMS lid completes the device.
The microchannels are 100 m side, 5 mm long. The
electrodes patterned in tripolar configuration have 100 m x
100 m recording sites within the channels.
Figu
re 2. Schematic view of a microchannel electrode implant for bladder
afferent recording. The channels are typically 100m side.
B. Surgery
All animal procedures were performed in accordance with
the Animals (Scientific Procedures) Act 1986. Adult
Sprague-Dawley rats (n = 14 at 300g) were anaesthetized
with either isoflurane for recovery procedures, or urethane
for terminal procedures. A laminectomy of the T11-L1
lumbar vertebrae was performed, and the dura incised
exposing the L5 and L6 dorsal roots. The left L6 dorsal root
was cut 3 mm caudal from the dorsal root entry zone. The
end of the transected nerve root was held with fine forceps
and carefully teased apart caudally into smaller strands along
its length, until a suitable size (100 m).
C. Implantation
Five teased strands of the unilateral L6 root were laid in
five PDMS micro-channels. A PDMS lid was placed on top
of the array, insulating the rootlets. Ultrasound images were
taken pre surgery and at 3, 7, 14, and 28 days post surgery
with a MicroMaxx (Sonosite). Animals were sacrificed at 7
(n = 2), 14 (n = 2), 28 (n = 6) days after implantation, and
bladder and dorsal root with ganglia removed for
histological assessment (Haematoxylin and Eosin, !-mouse
NF200 (1:200, Abcam), !-rabbit Vimentin (1:1000, Abcam).
D. Electrophysiology
Teased strands of the L6 root were laid on two platinum
hook electrodes. The bladder of the rat was catheterized
through the urethra, and connected to either a column of
water or a 0.5 ml syringe via a pressure transducer
(Digitimer). The bladder was distended through manual
infusion of the bladder (0.5 ml bolus), or hydrostatic
pressure increments: 10cm-70cm. Differential recordings
were conducted with a Neurolog amplifier and filter
modules (Digitimer), sampled at 43 kHz and digitized with a
Micro 1401 analog-to-digital converter (Cambridge
Electronic Design).
IV. RESULTS
A. Teased rootlets
The ipsilateral L6 dorsal root is isolated from the spinal
cord after laminectomy and transected near the dorsal root
entry zone. The root is then gently teased apart into rootlet
strands of approximately 100 m diameter and laid in the
open PDMS microchannels. Note that it is important to keep
the tissue immersed in extracellular medium at all times
during the procedure to ensure the nerve strands remain
functional.

Figure 3. Optical micrograph of L6 dorsal root teased in ! 100 m
strands and inserted in a passive array of PDMS microchannels.
Placing a PDMS lid on top of the device then closes the
250

channel array. Figure 3 shows a top view of the teased
rootlets in the channel array.
B. Bladder function
Bladder function is assessed using ultrasound across 28
days following unilateral L6 dorsal root implantation.
Specifically, the bladder volume is determined by measuring
width, depth and length of the bladder from the ultrasound
scans in nave, SCI and root implanted rats. SCI leads to
retention of urine due to neurogenic bladder (vol. = 1.83
cm
3
)

compared to the nave cases (vol. = 0.13 cm
3
). Figure 4
presents the change in bladder volume as a function of post-
implantation days: the volume is < 0.1 cm
3
at all times
indicating no urine retention in rats implanted with the
passive PDMS channel arrays.

Figure 4. Measured bladder volume over time in PDMS
microchannel implanted rats.
However bladder histology on rats with unilateral dorsal
root transection and implantation of the PDMS array shows
that the bladder detrusor muscle (including outer smooth
muscle, lamina propria, inner epithelial layer) becomes
thinner. The muscle atrophy is also accompanied with dome
expansion of the bladder.
C. L6 root interface
At 28 days post-implantation, L6 dorsal root histology is
conducted. Figure 5 presents a macroscopic view of the
explanted implant connected to the L6 dorsal root ganglion.

Figure 5. Optical picture of teased L6 rootlets with PDMS channel
implant at 28 days post-implantation.
The dissection of the implant reveals rootlets have remained
within the micro-channels and are surrounded with scar
tissue. Immunohistochemistry of the rootlets within the
channels shows axons are still present in the implant after 28
days. Scar tissue is clearly apparent at the distal and
proximal ends of the implant (Fig. 5) and connective tissue
is observed surrounding the primary afferent axons within
the implant.
D. Bladder afferents recording
Bladder afferent signals are monitored using Pt hook
electrodes. The recording is conducted in oil to maximize
the AP extracellular signal. During increasing hydrostatic
pressure infusion of the bladder, afferent activity can be
recorded representing distension of the bladder (Fig. 6).

Figure 6. Correlation between bladder filling (top trace) and
bladder afferent activity (bottom trace) recorded under oil with
hook electrodes on the teased L6 dorsal root.
After the bladder is filled (0.5mL), across a period of 60
minutes, a micturition pressure pattern can also be recorded
alongside bursts of afferent activity, showing different action
potential waveforms. Bladder contractions are intermittent
pulses, and correlate with large action potentials. A baseline
bladder activity underlies it.
No action potentials are recorded from the L6 rootlets
using the Pt hook electrodes when the experiment is
conducted in saline environment.
PDMS electrode implants with teased L6 dorsal rootlets
are then assessed in acute experiments. A teased rootlet is
laid manually into a 100 m side PDMS channel which is
then closed with the PDMS lid. The experiment is
conducted under oil at 37 C.
Figure 7a illustrates the experimental set-up. The
electrodes are in a quasi-tripolar configuration with the two
outer electrodes shorted together to one input of the
amplifier, and the recording electrode connected to the
second input of the differential amplifier. In this particular
design, the quasi-tripole is unbalanced with the central
electrode located 1/3 2/3 from each end electrodes. The 40
nm thick gold electrodes encapsulated in PDMS have an
impedance of ~ 100 k" at 1kHz.
Fig. 7b illustrates an extracellular recording with the
microchannel electrodes. Despite the surrounding noise, up
to 10 V amplitude APs are clearly detected. Surrounding
noise is likely to be due to electromyogram (EMG)
interferences and electrode Johnson noise.
No direct correlation between the recorded APs and
bladder filling was observed. However, because the L6
dorsal root contains afferents of the skin from the tail and
hindpaw, muscle spindle, as well as the bladder, burst of
activity was recorded when the rats paw was lightly pinched
251

during about 10 seconds as shown Fig. 7b.

Figure 7. (a) In situ picture of a PDMS electrode
microchannelimplant with teased L6 rootlet. (b) Afferent signals
recorded from mid-electrode in PDMS channel in pseudo-tripolar
configuration. Pinch indicates stimulation of the rat dermatome.
V. DISCUSSION
Unilateral transection, teasing and implantation of the L6
root within a PDMS microchannel interface lead to smooth
muscle atrophy of the bladder but no functional deficit in
bladder physiology. An alternative to sectioning the root to
position the implant is to tease the intact root. Once the
teased rootlets are separated, the open channel PDMS
implant is gently placed underneath the rootlets and closed
with the PDMS lid. This is a surgical challenge in particular
in a rat model but evaluation of functional and
morphological bladder alterations is on-going.
At one month post-implantation, primary afferents remain
within the implant and are fully functional as shown by the
AP recording with Hook and microchannel electrodes. Both
recordings were however conducted under oil, which
suggests that the PDMS microchannels were not sealed
electrically: the electrical impedance of the medium in the
immediate vicinity of the teased rootlet was therefore very
low and extracellular signals were not detectable. Using an
alternative fabrication process where a single PDMS
microchannel embeds gold thin film electrodes connected in
a tripolar configuration, we have however successfully
detected APs under saline environment from L5 teased
dorsal root [9]. The closed PDMS channel was assembled
by plasma-bonding the PDMS channel roof to the electrodes
on PDMS; the teased rootlet was subsequently pulled
through the channel using a nylon thread. We are now
exploring alternative process to seal the channels after
surgery.
Bladder afferents can be reliably recorded in vivo from
hook electrodes under oil but not from the microfabricated
electrodes. A Hook electrode under oil is the ideal
microchannel electrode as the low impedance electrodes are
within a channel formed by insulating oil around the
teased rootlet. Current PDMS microchannel electrodes need
to be improved in terms of (i) electrical impedance their
impedance is 2 to 3 orders of magnitude higher than that of
Hook electrodes hence their Johnson noise is of the order of
the AP amplitude, and (ii) microchannel assembly the
positioning of the PDMS lid on the open top channels is
prone to leakage. Efforts are underway to reduce further the
impedance of the gold on PDMS electrodes and optimize the
implant assembly process keeping in mind microfabrication,
geometrical, biological and surgical constraints.
REFERENCES
[1] E. A. Tanagho, "Urologic complications of spinal cord injury,"
Urology Clinical N Am, vol. 20, pp. 453-464, 1993.
[2] G. S. Brindley, "An implant to empty the bladder or close the
uretha," Journal of Neurology, Neurosurgery and Psychiatry,
vol. 40, pp. 358-369, 1977.
[3] G. S. Brindley, et al., "Sacral anterior root stimulator for bladder
control in paraplegia," Paraplegia, vol. 37, pp. 28-32, 1982.
[4] M. Craggs, "Chapter 61. Restoration of complete bladder
function by neurostimulation," in Textbook of the neurogenic
bladder, 2002, pp. 704-716.
[5] J. FitzGerald, et al., "Microchannels as axonal amplifiers," IEEE
Transactions on Biomedical Engineering, vol. 55, pp. 1136-
1146, 2008.
[6] S. P. Lacour, et al., "Flexible and stretchable micro-electrodes
for in vitro and in vivo neural interfaces," Medical Biological
Engineering and Computing, vol. 48, pp. 945-954, 2010.
[7] J. FitzGerald, et al., "Microchannel electrodes for recording and
stimulation: in vitro evaluation," IEEE Transactions on
Biomedical Engineering, vol. 56, pp. 1524-1534, 2009.
[8] J. FitzGerald, et al., "Recording with microchannel electrodes in
a noisy environment," in 30th Annual International IEEE EMBS
Conference, Vancouver, 2008, pp. 34-37.
[9] I. Minev, et al., "An elastomer microelectrode array for
recording afferent nerve activity in an anaesthetised rat,"
submitted, 2011.


252