UNCORRECTED PROOF

J. Neural Eng. 0 (2022) xxxxxx https://doi.org/10.1088/1741-2552/ac8ed6

Journal of Neural Engineering

PAPER

Liquid crystal electro-optical transducers for electrophysiology

RECEIVED
12 May 2022
REVISED
sensing applications Q1
Q2
18 August 2022
ACCEPTED FOR PUBLICATION
Amr Al Abed1,∗, Yuan Wei2,5, Reem M Almasri1,5, Xinyue Lei2, Han Wang1, Josiah Firth3, Yingge Chen2, Q3
1 September 2022 Nathalie Gouailhardou1,4, Leonardo Silvestri2, Torsten Lehmann2, François Ladouceur2 and Nigel H Lovell1,∗
1
PUBLISHED Graduate School of Biomedical Engineering, UNSW, Sydney, NSW 2052, Australia
xx xx xxxx 2
School of Electrical Engineering and Telecommunications, UNSW, Sydney, NSW 2052, Australia
3
Australian National Fabrication Facility, UNSW, Sydney, NSW 2052, Australia
4
School of Medical Sciences, UNSW, Sydney, NSW 2052, Australia
5
Authors contributed equally.
∗
Authors to whom any correspondence should be addressed.
E-mail: amra@unsw.edu.au and n.lovell@unsw.edu.au

Keywords: optrode, sensor, optical interfaces, liquid crystal

Q4
Supplementary material for this article is available online Q5

Abstract
Objective. Biomedical instrumentation and clinical systems for electrophysiology rely on electrons
for sensing and transmission of signal from the biological material. However, this electronic
approach constrains bandwidth, signal conditioning circuit designs, and the number of channels in
invasive or miniature devices. This paper demonstrates an alternative approach using light to sense
and transmit the electrophysiological signals. Approach. We develop sensing, passive,
fluorophore-free optrode based on the birefringence property of liquid crystals (LCs) operating at
the microscale. Main results. We show that these optrodes can have the appropriate linearity (µ ±
s.d.: 99.4 ± 0.5%, n = 11 devices), relative responsivity (µ ± s.d.: 57 ± 12% V−1 , n = 5 devices),
and bandwidth (µ ± s.d.: 11.1 ± 0.7 kHz, n = 7 devices) for transducing electrophysiology signals
into the optical domain. We report capture of rabbit cardiac sinoatrial electrograms and
stimulus-evoked compound action potentials from the rabbit sciatic nerve. We also demonstrate
miniaturisation potential by fabricating multi-optrode arrays, by developing a process that
automatically matches each transducer element area with that of its corresponding biological
interface. Significance. Our method of employing LCs to convert bioelectric signals into the optical
domain will pave the way for the deployment of high-bandwidth optical telecommunications
techniques in ultra-miniature clinical diagnostic and research laboratory neural and cardiac
interfaces.

1. Introduction as the development of materials for improved tissue-

Q6 Over the last century the field of electrophysiology has
primarily relied on arrays of metal electrodes for spa-
tiotemporal mapping of cardiac rhythms [1–3], inter-
rogation of neural circuits [4, 5], and brain machine
interfacing [6]. This ‘wire’ based technology is at the
core of all commercially available and research-grade
multi-electrode arrays (MEAs) [7, 8] as well as clin-
ical cardiac rhythm and neurological systems. Recent
progress include advances in complementary metal-
oxide-semiconductor (CMOS) design and microfab-
rication, which has enabled MEAs with high channel
count and a high channel packing density [4], as well
device interfaces [9]. However, the introduction of
high-resolution or high-channel count variants of
these arrays in healthcare diagnostics and implantable
bionics has been stalled by the challenge of hermetic
encapsulation, packaging and managing the multi-
tude of leads required to transfer signals from the bio-
logical interface to data processing systems.
Another widespread, albeit research laboratory-
confined, methodology for spatiotemporal interrog-
ation of electrophysiology signals is optical voltage
and calcium imaging [10, 11]. The last decade
has witnessed considerable advancement in this
field, including the development of fluorinated,

© 2022 IOP Publishing Ltd

J. Neural Eng. 0 (2022) xxxxxx A Al Abed et al

Figure 1. Optrode transducers. (A), (B) Illustration of operating principle of electro-optical transducers utilising deformed helix
ferroelectric liquid crystals. (A) Microscopic view of liquid crystal arrangement in a helix structure and the light polarisation state
changing with the applied electric field. (B) Schematic of the deformed helix structures of liquid crystals, represented by cylinders,
in the presence of electric fields, e.g. E = 0, E > 0 and E < 0, and corresponding rotation, Ω(E), of the optical axes associated
with the ordinary (no ) and extraordinary (ne ) refractive indices. ⊗ and ⊙ represent the electric field direction out of and into the
page, respectively. Red line represents the orientation of the molecular axis. (C) Representative microscopy images showing an
example of unaligned and aligned liquid crystal domains. At least one image was taken per fabricated device. Scale bars, 20 µm.

long wavelength, and photoacoustic voltage-sensitive
dyes [12], flourophores based on membrane poten-
tial dependent photoinduced electron transfer [13],
as well as genetically encoded voltage indicators
[14–16]. However, these methodologies do have
drawbacks; fluorophores have to be added to the bio-
logical preparation or virally transfected into cells and
genetically expressed, signals are read out as relative
changes in voltage with no routine means for quan-
tifying the absolute membrane potential, and photo-
bleaching and/or photo-toxicity limit the duration of
experiments.
One solution that can overcome these lim-
its is utilising photonics for detection and higher-
bandwidth transmission of electrophysiology sig-
nals. Specialised deformed helix ferroelectric (DHF)
liquid crystals (LCs) can smoothly, continuously and
passively transduce electrical signals into the optical
domain thus affording all the advantages typically
associated with optical communications networks
[17, 18]. So far, these DHF-LC transducers have
been employed in industrial applications [19–22],
sensing signals in the order of volts rather than
the submillivolt amplitudes typically encountered
in extracellular electrophysiology applications, and
encoding digital signals, rather than feature-rich time
varying waveforms of cardiac electrograms or neural
compound action potentials (CAPs) and local field
potentials.
We hypothesise that a passive fluorophore-free
optrode based on these DHF-LCs can transduce elec-
trophysiological signals. We have previously presen-
ted a theoretical model of the sensing optrode’s
operation principle [23]; in this paper, we valid-
ate its applicability to electrophysiology through sys-
tem characterisation and performance assessment of
single channel optrode devices in capturing cardiac
electrograms and nerve CAPs, as well as through the
fabrication of multi-optrode arrays (MOAs).
1.1. Theory: the optrode’s mechanism of operation
In brief, when no electric field is applied, the aver-
age orientation of the DHF-LC molecules sand-
wiched between two conductive substrates exhibits
a helical structure whose axis lies in the cell’s plane
(figures 1(a) and (b)) and whose orientation in that

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J. Neural Eng. 0 (2022) xxxxxx
plane is dictated by alignment layers (figure 1(c))
deposited on the inner surfaces of the substrates. If
an electric field is applied perpendicularly to the hel-
ical axis, the molecular dipole moments will tend to
align with it, thus deforming the helical structure. Still
on the macroscopic scale, this deformation produces
a rotation of the optical axes by an angle which, for
small fields (e.g. <0.1 V µm−1 ), is proportional to the
strength of the applied electric field.
A functional electro-optical transducer exploiting
this effect can be realised by attaching a linear polar-
iser and a mirror to the external surfaces of the front
and back substrates, respectively. In this arrangement,
a light beam from the front is reflected back by the
mirror passing twice through the linear polariser and
the LC material, so that its intensity is modulated by
the electric field applied to the DHF-LC. When the
optical axis of the linear polariser is at the optimal
angle (≈23◦ ) relative to the optical axis of DHF-LCs
at no external electric field, the intensity of the reflec-
ted light can be made proportional to the rotation
of the optical axes. From this baseline configuration,
the DHF-LC optical axis, and in turn, reflected light
intensity, is proportional to the strength of the elec-
tric field across the two layers, thus producing a linear
electro-optical response. This forms the basis of our
optical-electrodes ‘optrodes’. This transduction pro-
cess is entirely passive in nature.
1.2. Terminology
The following terminology will be adopted (figure 2).
A construct of DHF-LCs sandwiched between a con-
ductive transparent layer and a conductive reflective
layer is referred to as ‘transducer’. We use the term
‘device’ to refer to a transducer integrated with optical
components for light input/output and metallic pins
for electrical connection. Device nomenclature will
start with a D or I prefix, identifying the source
of LC cells used during fabrication, followed by a
single digit DHF-LC identifier. ‘Recording system’
is used to refer to a device and all external com-
ponents required to operate it, including photonic
and electronic parts for generating and transmitting
light as well as photodetection circuitry and data
acquisition instrumentation. ‘Multi-optrode array’ or
‘MOA’ is used to refer to a group of transducers on a
substrate.
2. Methods
2.1. Fabricating single channel optrode devices
Electro-optical transducers were fabricated by sand-
wiching a 5 µm thick layer of DHF-LC between two
≈0.7 mm thick indium tin oxide (ITO)-coated glass
substrates of empty LC cells sourced from Instec
(USA, devices prefixed with an I) and Depo Hi-Tech
(China, devices prefixed with a D), using glass spacer
rods and UV-curable glue [17, 18] (figure 2(a)).
The DHF-LCs were aligned with the aid of rubbed
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A Al Abed et al
planar-aligning polyimide (60 nm) on the inner face
of the ITO layers (KPI-300B and PI2016 polyimide
for transducers fabricated using Instec and Depo cells
respectively). A gold mirror was attached to the exter-
ior face of one ITO-glass substrate. Gold was chosen
as a reflective layer due to its excellent reflectance
(≈98%) at 1.55 µm. The ITO layers were connected
to metal pins for the application of electric poten-
tials. Polarised light (1550 nm) was delivered using
a polarisation maintaining (PM) fibre with a 10.5
µm diameter core (Flyin Optronics, China), via a
Gradient-Index lens and quarter-wave plate before
passing through the DHF-LC transducer. The light
was reflected back through the DHF-LC layer by the
transducer’s rear-mounted mirror and coupled back
into the same PM fibre. The diameter of the collim-
ated light beam projected on and reflected from the
mirror was ≈0.6 mm, defining an area of ≈0.28 mm2
responsible for electro-optical transduction. The PM
fibre was designed to propagate only one polarisation
of the input light due to the strong birefringence and
different propagation constants of the two polarisa-
tion modes produced by inducing stresses in the fibre.
Therefore, the PM fibres worked both as a polariser
and an analyser in this work. The angle between the
slow axis of the PM fibre and the helix axis of the
DHF-LC was ≈23◦ , which was chosen to have the best
linear optical response [18].
The PM optical fibre setup was advantageous
compared to an unpolarised setup reported for indus-
trial applications [17, 18]. Firstly, the need for a separ-
ate polariser is eliminated, which reduces the number
of components. Additionally, the setup is less sensitive
to fibre bending and to vibrations in general, which
can increase the signal to noise ratio (SNR) in prac-
tical circumstances.
2.2. Integration of a quarter-wave plate into
optrode devices
Theoretically, the optical response of optrode trans-
ducers after the light beam double-passes through
the DHF-LC layer is determined by several paramet-
ers, including the LC thickness d, electric field con-
trolled birefringence ∆n(E), and electric field con-
trolled optical axes rotation Ω(E) according to the
expression of the parallel reflectance (R∥ ) as
 
2πd∆n(E)
R∥ = 1 − sin 2
sin2 [2β − 2Ω(E)] (1)
λ
where λ is the operation wavelength and β is the
angle between the linear polariser and the optical axis
of DHF-LCs with no externally applied electric field.
The external electric field can change the polarisa-
tion state of incident light via both electric field con-
trolled birefringence ∆n(E) and optical axes rotation
Ω(E) of DHF-LCs. As a result, the optical response of
DHF-LC transducers R∥ is composed of a background
DC component and the optical response containing
J. Neural Eng. 0 (2022) xxxxxx A Al Abed et al

Figure 2. An optrode recording system for electrophysiology. (A) Schematic cross section and photo of a single channel optrode
device. Schematic dimensions are not to scale. (B) Block diagram of the optrode recording system. A fibre-based optical setup is
used to deliver light to the optrode device. The optrode transduces biopotentials into a light signal, which is then routed to a
photodetector for conversion into an electrical signal and acquisition. (C) Schematic illustration of the rabbit in vivo sciatic nerve
experimental setup for the sensing optrode and conventional bioamplifier systems. A similar setup was used for ex vivo cardiac
electrophysiolgy experiments except that no stimulation was required as the tissue was spontaneously pacing. PM,
polarisation-maintaining; GRIN, gradient-index; ITO, indium tin oxide; SLD, super-luminescence diode; TEC, thermoelectric
temperature controller; DAQ, data acquisition system.

detected time-variant signals with the information
of interest. As discussed later, the relative responsiv-
ity of optrode transducers is measured as the frac-
tional change of reflectance per volt, which illus-
trates the ability of DHF-LCs to modulate light with
time-variant signals relative to the background DC
component. Therefore, the relative responsivity of
optrode transducers is limited by the background DC
component especially when the value of this DC com-
ponent is greater than the amplitude of time-varying
optical responses. To optimise the relative respons-
ivity of optrode transducers by reducing the back-
ground DC component, the quarter-wave plate con-
verts the linear polarisation into elliptical or circular
polarisation before transmission through the DHF-
LC layer. With the phase retardation introduced by
the quarter-wave plate, the background DC com-
ponent of reflected light intensity can be minimised
without a significant reduction in the amplitude of
time-varying optical responses [24].

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J. Neural Eng. 0 (2022) xxxxxx
Figure 3. Process diagram for the fabrication of multi-optrode arrays. UV, ultra-violet; DRIE, deep reactive-ion etching; PECVD,
plasma-enhanced chemical vapor deposition; ITO, indium tin oxide; DHF-LC, deformed helix ferroelectric liquid crystals.
2.3. Fabricating MOAs
Trackless MOAs were fabricated using a similar archi-
tecture to the single channel optrode device described
above, but the back substrate with the mirror was
replaced with a more complex structure. This process
is illustrated in (figure 3). We also fabricated an MOA
with embedded tracks to facilitate electrical poten-
tial application and device impedance characterisa-
tion. Details of the fabrication processes for MOAs are
detailed in the Supplementary Materials.
2.4. Microscopy assessment of optrodes
Still images (1280 px × 960 px) of the DHF-LC
domains were captured using an Eclipse LV100POL
(Nikon, Japan) microscope with a ×10 Nikon LU Plan
Fluor 10×/0.3 WD 17.5 NA objective and a polar-
iser via a DS-Fi1 camera (Nikon) using the NIS-
Elements F 2.30 imaging software (Nikon). Time-
series line scans of the DHF-LC was conducted using
a Leica TCS SP5 white light laser STED confocal
microscope with a ×10 0.3 NA objective and an
emission 670 nm bandpass filter. A 90◦ polariser
5
A Al Abed et al
was used to obtain optimal electro-optical transduc-
tion and hence response intensity. Responses were
detected using a photomultiplier tube (665–675 nm)
and acquired using the Leica Application Suite
Advanced Fluorescence software (Leica Microsys-
tems, Germany).
2.5. Benchtop setup for optrode device
characterisation
Polarised light was delivered to optrode devices,
via PM fibres, from a super-luminescence diode
(SLD) (1543.6 ± 58 nm, Anritsu, Japan) controlled
(200 mA, 25 ◦ C) using a CLD1015 compact laser
diode driver (Thorlabs, USA), passing first through
a PM optical isolator, attenuator (to 90%), and a
PM circulator (Flyin Optronics, China). Light reflec-
ted by the optrode’s mirror was guided through the
PM circulator to a germanium DC-coupled detector
(PDA50B2, Thorlabs, USA) with gain set to 20 dB.
Voltage waveforms were applied to the optrode from
a USB-6361 acquisition device (National Instrument,
USA) controlled using LabVIEW software v18 (NI,
J. Neural Eng. 0 (2022) xxxxxx
USA). Custom LabVIEW, MATLAB 2021a (Math-
works, USA), and R 3.6 [25]/RStudio 1.2 (RStu-
dio, USA) scripts were written for the analysis of
benchtop, SNR, and noise floor measurements.
2.6. Relative responsivity as a transducer metric
To compare optrode transducers, we define a quantity
R0 , expressing the relative change in reflectance per
applied volt, or
1 dR
R0 ≡ × 100%, (2)
R0 dV V=0
where R0 is the reflectance when 0 volts is applied.
This quantity, which we call the relative responsivity,
is an intrinsic property of optrode transducers and it
is inversely proportional to the system’s sensitivity, so
that optrode transducers with larger relative respons-
ivity values exhibit a larger electro-optical conversion
factor.
We propose the relative responsivity as a metric
for comparing transducers following the justification
detailed next. The reflectance as a function of the
voltage applied across the optrode, R(V), is S-shaped,
with the linear part of the curve centred around 0 V
and saturation regions at higher voltages, of the order
of ±1 V. This response can be measured by applying
a low frequency square wave AC voltage at various
amplitudes to the optrode and measuring the intens-
ity of the reflected light. Since the reflectance cannot
be negative, it follows that even when 0 V is applied
there is a certain amount of reflected light, i.e. R0 =
R(0) > 0. In this situation the system’s noise is given
to a good approximation by the sum in quadrature
of the photodetector’s noise and of the noise from the
light source. In order to maximise the SNR our system
was designed so that the latter dominates and it is pro-
portional to R0 , so that ∆Rnoise ∝ R0 . When a small
voltage, ∆V, is applied across the optrode, the reflect-
dR
ance changes by ∆R = dV V=0
∆V. The sensitivity of
the system is given by the smallest voltage ∆Vmin cap-
able of producing a signal ∆R at least equal to the
noise floor, ∆Rnoise . Based on our previous assump-
tions we find ∆Vmin ∝ R0 / dV dR
V=0
. It is therefore
convenient to compare optrode transducers using a
relative change in reflectance per applied volt metric,
i.e. R0 .
In practice, to obtain the relative responsivity a
square wave signal (5 Hz, ±0.5 V) was applied across
optrode devices and R0.5 , R−0.5 , and R0 were meas-
ured as the photodetector output at 0.5 V, −0.5 V, and
0 V, respectively. The reported relative responsivity
was calculated as
1 R0.5 − R−0.5
R0 = × 100%. (3)
R0 1V
2.7. Measuring device reflection
We calculated the reflectance of each optrode device
by measuring the ratio of light power delivered to and
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A Al Abed et al
reflected from the transducer. The DC-coupled pho-
todetector firstly recorded the signal when the trans-
ducer was shortened. Then, we replaced the trans-
ducer with the photodetector and recorded the light
signal at port 2 (figure 2(b)). In both situations, the
DC level of the recorded signals represents the light
power. The reflectance (R) was therefore calculated as
P3 V3
R≡ = (4)
P2 V2
where P3 and P2 are the light power at port 3 and port
2 of the circulator, respectively, and, V 3 and V 2 are the
measured DC voltages at these light powers.
2.8. Measuring device linearity
We applied sinusoidal waveforms (0 V to ±5 mV in
increments of ±0.1 mV, and 10 mV to ±100 mV in
increments of ±5 mV) at a frequency of 10 Hz. Each
test signal was applied for 2 s with 1 s of pre- and
post-baselines. The order of amplitudes was random-
ised. The output (×100 gain) was attenuated ×100 by
a resistor-divider to produce the desired amplitude.
Responses were measured with a sampling rate of
200 kHz and the acquisition input range was auto-
matically set to one of two levels (10 V or 500 mV)
depending on the applied waveform amplitude. For
a given test signal, the output was calculated as the
amplitude of a sine function fitted to the ensemble
average of response cycles.
Linearity is defined in terms of nonlinearity (%):
∆Vin(max)
nonlinearity = × 100% (5)
Vin(f.s.)
where ∆Vin(max) is the maximum input deviation
from the best fit linear regression model to the
measured data, and Vin(f.s) is the maximum, full-
scale input. Because the photodetector operates as
a reversed biased photodiode, which theoretically,
exhibits a linear response, the measured nonlinearity
of the transducer-photodetector system can be attrib-
uted to the optrode.
2.9. Frequency response analysis
The frequency response of the optrode transducer was
measured by applying sinusoidal voltage waveforms
(10 Hz–50 kHz, ten points per decade), to estimate
the 3 dB bandwidth. Because the DC coupled pho-
todetector has a bandwidth of 85 kHz, the measured
frequency response approximates that of the optrode
transducer.
2.10. Transfer function optimisation
The optrode’s transfer function was fitted to meas-
ured Bode plots using custom-written MATLAB
2021a scripts employing a parameter-constraining
version [26] of MATLAB’s implementation of the
Neldler-Mead Simplex optimisation algorithm [27].
J. Neural Eng. 0 (2022) xxxxxx
Model parameters were estimated by minimising the
following objective function:
X oidal waveform by the variance of 1 s of recording
obj(f ) = [(ℜmodel − ℜmeasured )2
+ (ℑmodel − ℑmeasured )2 ] (6)
where ℜ (Ω) and ℑ (Ω) are the numerically-derived
real and imaginary parts and f (Hz) the frequency.
2.11. An optrode recording system for
electrophysiology
The optrode system setup is shown in figure 2(b).
Polarised light was generated by a 1543.6 ± 58 nm
SLD (Anritsu, Japan) controlled using a compact laser
diode driver (CLD1015, Thorlabs, USA) with tem-
perature control (500 mA, 20 ◦ C). Output light is
passed through a PM optical isolator (Flyin Optron-
ics, China) to protect the SLD from reflected light,
attenuated (to 5%) by 10/90% and 50/50% PM split-
ters (Flyin Optronics, China), guided through a PM
optical circulator (1310–1550 nm, Flyin Optronics,
China) to the optrode transducer. Light power used
in experiments was ≈600 µW incident on the optrode
transducer. Reflected light is then guided and isolated
by the third port of the optical circulator to an AC
coupled custom-built photodetector board, consist-
ing of an InGaAs PIN diode (Flyin Optronics, China),
trans-impedance amplifier, and a series of band-pass
filters (0.1 Hz–20 kHz).
Adjustments from the benchtop characterisation
setup were required because electrical and optical
characterisation needs the DC component of the out-
put signal (for example to measure linearity and sens-
itivity). Whereas in biological recording, only the AC
output signal is of interest so an AC coupled photo-
detector is preferable so that the gain and light power
can be set to higher values in order to increase the
SNR without DC interference or saturation of the
photodetector.
2.12. Benchtop characterisation of the optrode
recording system
The optrode system (figure 2(b)) gain was meas-
ured by applying a sinusoidal waveform (50 mV,
99 Hz) from a data acquisition system (PowerLab
35/4, ADInstruments, New Zealand) to each optrode
device and recording responses. The gain measure-
ment was conducted prior to each experiment.
The optrode system signal quality was investig-
ated by applying a sinusoidal waveform (0.5 V, 99 Hz)
via a pair of 1 mm diameter platinum hook electrodes
into 0.9% Phosphate-buffered saline (PBS) solution,
and recording responses using a pair of 1 mm dia-
meter platinum electrodes positioned inline 1 cm
from the stimulating electrodes, in reference to a
Ag/AgCl electrode placed in the PBS. Responses were
also recorded via a conventional bioamplifier sys-
tem (CardioPhys, World Precision Instruments, USA,
0.1 Hz–10 kHz). A sinusoidal waveform was fitted to
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A Al Abed et al
1 s of the response to the test signal. The SNR was cal-
culated by dividing the variance of the fitted sinus-
with no test signal applied (baseline noise).
The noise floor for the optrode and for a con-
ventional bioamplifier recording systems were ana-
lysed from 1 s of baseline recording, via a pair of
1 mm diameter platinum hook electrodes immersed
into PBS solution. In the time-domain, the noise floor
was estimated by computing the standard deviation
of the signal (Vs.d. ) as this metric is immune to DC
offsets. To generate the power spectrum density, we
bin-averaged the measured noise floor data in the fre-
quency domain. The size of each bin increases logar-
ithmically so that the data points are evenly distrib-
uted over the frequency axis.
Recovery from the stimulus artefact was investig-
ated. Pulses (0.1 V–10 V, 50 µs–500 µs) were applied
to an exposed rabbit sciatic nerve postmortem via a
pair of 1 mm diameter platinum hook electrodes and
responses recorded via a pair of 1 mm diameter plat-
inum electrodes positioned 1 cm distally.
For all system characterisation, both optrode and
bioamplifier signals were low-pass filtered (25 kHz)
prior to digitisation (100 kHz, 16 bits, input range
±10 V) using the PowerLab data acquisition system.
2.13. Animal studies
From the seven I3 and the five D3 single chan-
nel optrode devices fabricated one device from each
group was used in all experiments.
2.14. In vivo sciatic nerve electrophysiology
New Zealand White rabbits (n = 4 females, age > 6
months, 3.1–3.4 kg) were anaesthetised by isoflur-
ane inhalation to effect up to 5% in 2 L min−1 O2 .
Heart rate, blood oximetry, and temperature were
continuously monitored as were reflexes to ascertain
surgical anaesthesia. Body temperature was main-
tained by placing rabbits over a heat mat and covering
with blankets. Hartmann’s physiological solution was
continuously supplied via an intravenous catheter
through the marginal ear vein (3–4 ml min−1 kg−1 ).
An incision was made through the skin and
muscles of either hind-leg to expose a section of the
right or left sciatic nerve (n = 2 each). Parafilm was
used to isolate the nerve from surrounding tissue and
the nerve was maintained moist by regular applica-
tion of paraffin oil, in effect creating a paraffin oil pool
immersing the nerve section of interest.
In-house fabricated 1 mm diameter platinum
hook electrodes were used for electrophysiology
recording and stimulation (figure 2(c)). Monophasic
constant-voltage stimulus pulses were generated by
a stimulus isolation unit (2200, A-M Systems, USA)
driven by an analog output from a PowerLab 35/4
acquisition system. Trains of 50 repeat pulses were
delivered (10 Hz, inter-train interval of 60 s). For each
J. Neural Eng. 0 (2022) xxxxxx
repeat, the pre-pulse baseline was 10 ms. Pulse amp-
litude (0.1 V–10 V) and width (50 µs–500 µs) were
randomised between pulse trains.
Stimulus evoked nerve responses were recor-
ded in a bipolar configuration (figure 2(c)). In the
case of bioamplifier recordings, a third hook elec-
trode acted as reference. Signals from the hook elec-
trodes were transmitted to either the optrode system
(gain 3.31 ± 0.17) or a conventional bio-amplifier
(gain 100, 0.1 Hz–10 kHz, CardioPhys). This pro-
tocol enables removal of the tissue interfacing as a
factor when comparing measurements obtained via
bioamplifier and optrode systems. Switching between
conventional bioamplifier and optrode recording sys-
tems was done after completion of an entire stimula-
tion protocol (≈30 min).
2.15. Ex vivo cardiac electrophysiology
Following in vivo experiments a sternotomy was per-
formed. Cardioplegia solution at 4 ◦ C was poured
into the chest cavity and the heart rapidly excised and
washed in cold Cardioplegia solution.
A sinoatrial node and surrounding right atrial
pectinate muscle tissue preparation was dissected in
Krebs-Ringer solution. The tissue was then mounted,
endocardial face up, in a modified RC-26 recording
chamber (Warner Instruments, USA) using fine pins
into the chamber’s sylgard floor (figure 5(a)), and
superfused with Krebs-Ringer solution, gassed with
carbogen (95% O2 , 5% CO2 ), circulated at a rate of
≈4 ml min−1 by two MP-II Mini peristaltic pumps
(Harvard Apparatus, USA). The temperature of the
superfusate solution was monitored and maintained
at 34 ◦ C by means of a TC-324B temperature control-
ler (Warner Instruments, USA).
Bipolar cardiac electrograms were captured by
means of a pair of a fine in-house fabricated tung-
sten electrode in reference to an Ag–AgCl ground
pellet placed distally in the recording chamber. Sig-
nals from the electrodes were sequentially transmit-
ted to either the optrode system (gain 27.553 ±
1.546) or a conventional bioamplifier (CardioPhys,
gain 100, 0.1 Hz–10 kHz). Similar to nerve electro-
physiology experiments, this protocol eliminates the
tissue interface as a factor when comparing measure-
ments obtained via bioamplifier and optrode systems.
2.16. Electrophysiology data analysis
Signals from both the optrode setup and bioampli-
fier were recorded using a PowerLab data acquisition
device (100 kHz, 16 bits, ±1 V) using LabChartPro 8.
For CAPs, the bioamplifier signal was filtered
prior to digitisation (2 kHz). For comparison, the
optrode signals were filtered using a software imple-
mented 2 kHz lowpass filter (MATLAB 2019b). The
baseline drift arising due to the recovery from the
stimulus artefact was estimated by fitting a two order
exponential function to the response in MATLAB
2019b,
8
A Al Abed et al
V(t) = (Vmin − a)ebt + aect + Vss , t ∈ [tmin , tend ]
(7)
where (tmin ,Vmin ) is the minimum response point and
Vss is the steady state baseline voltage following the
response, defined as the average of the final 30 ms
of recording. The baseline drift following the stimu-
lus artefact was then removed by subtracting the fit-
ted function from the response; a similar approach to
template-based stimulus artefact removal algorithms
[28, 29] (refer to supplementary figure 1 for justific-
ation of baseline function choice). This process was
repeated for each individual response.
Features of cardiac signals were detected using
LabChartPro’s ‘evoked responses’ peak detection and
peak-to-peak interval algorithms, where all values
were manually verified by an investigator. The amp-
litude of the CAPs and cardiac electrograms were
defined from the baseline to the highest point of
the peak. The baseline of the cardiac electrogram
was determined by the maximum diastolic potential,
while the baseline of the CAPs was the mean of the
first 10 ms pre-stimulus.
Statistical analysis was conducted in Prism 8
(GraphPad Software Inc., USA). Two-way ANOVA Q7
followed by Sidak’s multiple comparison test were
performed on the in vivo data to compare the optrode
recording performance to the bioamplifier based
on the response peak-amplitude and latency. Two-
tailed unpaired t-tests were performed to compare
the optrode cardiac recording performance to the
bioamplifier based on the peak amplitude, heart
rate, and slope of the diastolic phase. The normal-
ity assumption was passed for both ANOVA and t-
tests. Pearson correlation coefficients were obtained
by aligning the CAPs or cardiac waveforms obtained
from the bioamplifier with those obtained from the
optrode to compare their waveform morphometry
and measure their linear dependence. Signals were
aligned based on the timing of the peak of the first
time-derivative of the voltage signals to reduce the
effect of noise and baseline drifts. For the purposes
of statistical inference a value of 0.05 was used to test
the null hypothesis. A p-value less than 0.05 was con-
sidered statistically significant.
3. Results
We fabricated twelve single channel optrode devices
for assessing the effect of quarter-wave plate integra-
tion on device bandwidth and relative responsivity.
Additional I3 (n = 7) and D3 (n = n) optrode devices
were fabricated for more comprehensive benchtop
characterisation. One device was selected from each
of these two groups for evaluation in biological
experiments.
3.1. Sensor characterisation of optrode devices
Single channel sensing optrode devices were fab-
ricated (n = 12) using various combinations of
J. Neural Eng. 0 (2022) xxxxxx A Al Abed et al

Figure 4. Optrode benchtop characterisation. (A) Performance of optrode devices with different deformed helix ferroelectric
liquid crystals (digits) and alignment layers (D or I prefix) with (orange) and without (blue) a quarter-wave plate (n = 1 device
each). Devices that were further characterised are in bold. (B) Linearity of optrode devices used in biological experiments (◦) with
measurement root mean square error (RMSE) normalised to the input voltage (×). Non-linearity; D3, 0.03%, I3, 0.15%.
(C) Measured Bode plots for optrode devices (−) and the optrode recording system (◦) used in biological experiments with D3
(orange) and I3 (blue) transducers. (D) Bode magnitude and phase plots of optrode devices, as standalone units, from
measurements (blue) and fitted relaxation models (orange). (•) mean; I3, n = 7 devices; D3, n = 5 devices. Error bars, s.d.
(E) Stimulus artefact response from a 0.7 V and 200 µs pulse signal across a rabbit sciatic nerve postmortem for optrode
recording system with D3 (orange) and I3 (blue) transducers and bioamplifer system (gold). The inset shows the recovery from
the negative peak. The time course of the full recovery back to baseline is not shown. (F) Noise floor power spectrum density for
the optrode recording system using I3 (n = 7) and D3 (n = 5) devices in phosphate-buffered saline solution.

DHF-LCs and alignment layers, and their character-
isation undertaken. An inverse relationship between
relative responsivity and bandwidth can be noted
(figure 4(a)). The addition of the quarter-wave plate
to the optrode device design increased the relative
responsivity of electro-optical transduction signific-
antly (p < 0.001, n = 12 devices, Wilcoxon matched-
pairs signed rank test). The median improvement in
relative responsivity was 226% (inter-quartile range
174%–300%, range: 147%–585%).
Based on this initial assessment we identified
optrode devices I3 and D3 for in-depth charac-
terisation, as they displayed linear responses and
bandwidths appropriate for recording fast and slow
biopotential signals, respectively (table 1). These
devices contained the same DHF-LCs aligned using
two different alignment materials. For this purpose
seven I3 and five D3 single channel optrode devices
were fabricated and their electro-optical transduction
properties characterised.
We first considered the linearity of devices.
Ideally, the optical output of optrodes (represented
by the output voltage of a photodetector) should be
linearly related to the applied input voltage. In the
−0.1 to 0.1 V input range our measurements sup-
port this characteristic with an average nonlinearity

9
J. Neural Eng. 0 (2022) xxxxxx
Table 1. Performance metrics and transfer function (H) parameters for two types of optrode devices at room temperature. Mean ± s.d.;
n = 7 I3 devices and n = 5 D3 devices, with the exception of linearity measurements where one I3 device was excluded due to
mechanical failure. Standalone characterisation of optrode devices was conducted using a wide bandwidth photodetector. The system
included optrode devices as well as all optical and electronic components used in electrophysiology experiments as illustrated in figure 2.
PBS, Phosphate-buffered saline solution.
Property
Device Relative responsivity (% V−1 )
Reflectance (%)
Non-linearity (%)
Bandwidth (Hz)
Phase shift (◦ )
H gain
H characteristic frequency f 0 (Hz)
H fractional index β 1
H fractional index β 2
System Gain
SNR (dB) in PBS, Vinput : 0.5 V 99 Hz
Noise floor (Vs.d. ) in PBS (µV)
of less than 1% for both I3 and D3 devices (table 1,
figure 4(b)).
Having confirmed linearity of devices, we then
characterised their frequency response. The optrode
works as a low-pass filter when transducing electrical
signals to optical signals (figure 4(c)). Unlike a con-
ventional low-pass filter, it appears to introduce a
constant phase shift in the passband especially for
optrode device D3 compared to I3 (table 1). The fre-
quency response of the transducers can be fitted with
fractional order transfer functions. For I3:
G the recording system (figure 2(b)) were compared
H(f ) =   β1 β2
jf
1+ f0
and for D3:
G
H(f ) =  β 1  β 2
jf jf
1+ f0 + f0
where G is the gain constant, f is the frequency in
Hz, f 0 is the characteristic frequency constant, which
illustrates the transducer’s intrinsic relaxation speed,
β 1 and β 2 are the fractional indices, and j is the ima-
ginary unit. A population of optimised models is
generated for optrode devices (I3, n = 7, D3, n = 5,
table 1 for estimated parameters). We note that out
of all parameters, G has the highest variation, poten-
tially due to the current manual assembly process of
optrode transducers resulting in variations in reflect-
ance. The expression for I3 is in Havriliak–Negami
relaxation form [30]. Though this relaxation form
could also reproduce the measured Bode plots of D3
transducers, our derived empirical expression, which
is a variant from the Cole-Cole relaxation model [30],
provides a better fit (figure 4(d)). The goodness of
fit was assessed using the coefficient of determina-
tion. R2 values for I3 devices (n = 7) were 0.99 ±
0.0002 (magnitude) and 0.96 ± 0.02 (phase), and for
D3 devices (n = 5): 0.99 ± 5 × 10−5 (magnitude) and
10
A Al Abed et al
I3 D3
18 ± 4 57 ± 12
58 ± 26 11 ± 4
0.59 ± 0.41 0.55 ± 0.61
11 104 ± 672 620 ± 116
−0.15 ± 0.12 −4.78 ± 1.05
0.07 ± 0.03 0.37 ± 0.36
6.39 × 103 ± 438 1.01 × 103 ± 127
0.99 ± 0.02 0.14 ± 0.03
0.5 ± 0.04 0.92 ± 0.02
3±1 29 ± 9
−7.2 ± 16.1 6.6 ± 1.3
355 ± 89 84 ± 14
0.93 ± 0.01 (phase). Over the low frequency range
(<5 kHz), the model fitted the measured phase well
but deviated at higher frequencies potentially due to
measurement errors as the frequency approaches the
sampling rate.
3.2. Characterising an optrode electrophysiology
recording system
To investigate whether the photodetection circuitry
limited the overall bandwidth, the Bode plots of
optrode devices before and after integration with
(8) (figure 4(c)). The system has a minimal impact on
the magnitude Bode plot but introduced a signific-
ant phase shift, as expected due to the photodetec-
tion board’s 0.1 Hz–20 kHz bandpass filter. The lower
bandwidth of D3 devices 620 Hz, table 1) could be
(9) sufficient for recording cardiac electrogram but inap-
propriate for other applications such as capturing
nerve CAPs or signals of neural origin in general.
However, the larger bandwidth of I3 devices (11 kHz,
table 1) would be sufficient for most scenarios.
Considering many electrophysiological experi-
ments involve recording a stimulus-evoked responses,
we assessed as an example the stimulus artefact arising
from stimulating a nerve postmortem with a 0.7 V
and 200 µs pulse was quantified as an example
(figure 4(e)). The optrode system with either D3 or
I3 devices performed better at reducing the negative
deflection of the artefact compared to the bioamp-
lifier system (−1.7 mV, −3.0 mV, −3.7 mV, respect-
ively). The bioamplifier recording system reproduced
the artefact with the smallest peak (27.2 mV) fol-
lowed by the optrode recording system with the D3
(28.3 mV) and I3 (83.7 mV) devices. The recovery
of all systems converged at 5 ms post stimulus. The
optrode recording system with the D3 device exhib-
ited a slower recovery from the artefact compared to
the I3 device, which is consistent with their respective
bandwidth (table 1).
J. Neural Eng. 0 (2022) xxxxxx A Al Abed et al

Figure 5. Optrode sensing of cardiac electrograms. (A) Representative image of a cardiac sino-atrial node tissue preparation.
(B) Representative spontaneous waveforms obtained from rabbit sinoatrial node and surrounding right atrial pectinate muscle
tissue ex vivo, measured by optrode D3 (blue) and bioamplifier (orange) systems. An average of 50 waveforms is shown per
magnified panel.

Table 2. Cardiac electrogram metrics. Comparison of ex vivo cardiac electrogram features (mean ± s.d., n = 50 beats) measured via
optrode D3 and bioamplifier as well as the waveforms’ correlation coefficients. HR, heart rate; bpm, beats per minute; V˙d , slope of
diastolic phase; ρ, Pearson’s correlation coefficient.

Bioamplifier Optrode D3
No. HR (bpm) Amplitude (µV) V˙d (µV s−1 ) HR (bpm) Amplitude (µV) V˙d (µV s−1 ) ρ

1 170 ± 1 1579 ± 80 −265 ± 36 178 ± 4 1508 ± 142 −337 ± 100 0.96

2 169 ± 1 857 ± 48 857 ± 48 167 ± 2 642 ± 74 642 ± 74 0.86
3 172 ± 1 811 ± 7 811 ± 7 156 ± 2 933 ± 54 933 ± 54 0.98
4 163 ± 3 1187 ± 40 187 ± 112 159 ± 1 1199 ± 73 175 ± 102 0.99
5 160 ± 1 992 ± 4 731 ± 9 160 ± 3 1574 ± 41 849 ± 78 0.95
6 155 ± 1 2073 ± 12 200 ± 34 154 ± 1 2246 ± 45 224 ± 101 0.99
7 151 ± 1 270 ± 4 −65 ± 59 153 ± 8 402 ± 46 −9 ± 12 0.96
8 153 ± 1 651 ± 4 191 ± 10 154 ± 3 619 ± 46 27 ± 28 0.84
9 155 ± 1 436 ± 2 −119 ± 6 157 ± 1 582 ± 48 −154 ± 259 0.98
10 158 ± 1 921 ± 7 331 ± 16 158 ± 7 655 ± 43 184 ± 74 0.96

Finally to assess the baseline noise profile of this
new electrophysiological system, the SNR and noise
power spectra of the optrode system in PBS solution
is analysed (figure 4(f), table 1). For most of the band-
width (>5 Hz) the noise power spectrum density of
the system is <100 µV2 Hz−1 when recording with
either I3 (n = 7) or D3 (n = 5) optrode devices, and
attenuates at ≈20 kHz because of the low-pass filter-
ing by the photodetector circuitry. The noise profile
is consistent within each of the two device groups, but
varies between the two groups. This can be largely
explained by the different relative responsivity of I3
and D3 devices, confirming that the noise is domin-
ated by fluctuations in the light source’s intensity.
3.3. Optrode sensing of cardiac electrograms
The optrode recording system captured stable mono-
polar electrograms from ex vivo rabbit cardiac sino-
atrial tissue preparations. The baseline noise floor
of the optrode recording system with the I3 device
was Vs.d. = 230 ± 9 µV, which significantly reduced
the SNR compared to that with device D3 (see
supplementary figure 2 for example traces). For
subsequent analysis we focus on signals recorded
using optrode device D3.
Optrode D3 captured cardiac signal fea-
tures observed in the bioamplifier measurements
(figure 5), despite the relatively higher noise floor
(Vs.d. = 43 ± 3 µV) compared to that of the bioamp-
lifier (Vs.d. = 3 ± 1 µV). The spontaneous pacemak-
ing rate, amplitude, and slope of the diastolic phase
were statistically indistinguishable (p = 0.77, 0.82,
0.76, respectively, unpaired two-tailed t-test, n = 10
recordings from four tissue preparations, table 2).
Despite the variety of waveform morphologies cap-
tured, Pearson’s correlation coefficients indicate that
the cardiac waveforms measured by the optrode D3
and bioamplifier are strongly correlated (ρ > 0.84
for all n = 10, table 2). Overlaying signals, based on
the position of the maximum rate of change, revealed
a small phase shift of the D3 optrode signal during
the slow components of the non-diastolic portion
of cardiac waveforms compared to their bioampli-
fier counterparts. This could be due to the constant
phase shift in the passband of device D3 (table 1,
figure 4(d)).

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J. Neural Eng. 0 (2022) xxxxxx A Al Abed et al

Figure 6. Optrode sensing of stimulus-evoked nerve responses. The optrode system recordings were obtained using transducer I3.
(A) Representative compound action potentials recorded by the optrode (blue), and bioamplifier prior to (gold) and subsequent
to (orange) optrode recordings in response to indicated pulse intensities and widths. The average of 50 traces is shown per panel.
(B), (C) Pearson’s correlation coefficients, r, between the bioamplifier pre-optrode and optrode recordings (blue) and amplifier
pre- and post-optrode recordings (orange) at different stimulus pulse amplitudes (B) and widths (C). P = n.s., two-way ANOVA.
Error bars, s.d. (D) Stimulus intensity-response curve and (E), stimulus duration-response curve obtained via optrode recordings
(blue) in comparison with a conventional amplifier (orange). P = n.s., two-way ANOVA. Error bars, s.e.m. (F), (G) Response
latency at different stimulus pulse amplitudes (F) and widths (G) obtained via optrode recordings (blue) in comparison with a
conventional amplifier (orange). P = n.s., two-way ANOVA. Error bars, s.e.m. (B)–(G) Data points are means of four biological
replicates.

3.4. Optrode sensing of nerve CAPs
Optrode device I3 was selected based on benchtop
characterisation indicating it has the appropriate
bandwidth to record CAPs. Despite its lower SNR
in PBS solution compared to the bioamplifier sys-
tem, the optrode captured stimulus-evoked nerve
responses from the rabbit sciatic nerve in vivo
with an appropriate time response and sensitivity
(figure 6(a)).
There was a strong correlation between nerve
response waveforms recorded by optrode and
bioamplifier systems, as well as waveforms obtained
via the bioamplifier prior to and subsequent to
optrode recordings (Pearson’s correlation coeffi-
cients, ρ > 0.8, figures 6(b) and (c)). It is worth
mentioning that the two sets of control bioampli-
fier measurements were not identical (figure 6(a),
gold and orange waveforms). This can be explained
by the time elapsed (≈2 h) between measurements
which could lead to a deterioration in nerve vital-
ity or changes in tissue contact. Given that these
factors are also expected to come into effect during
the optrode measurements, any variations in wave-
form obtained using the optrode system and their

12
J. Neural Eng. 0 (2022) xxxxxx
bioamplifier counterparts can be considered insig-
nificant and not related to the optrode performance
itself.
Figures 6(d) and (e) demonstrate the ability of
the optrode system to record CAPs with similar amp-
litudes to those recorded by the bioamplifier sys-
tem, across different stimulus pulse intensities (p =
0.92, ANOVA, n = 4) and widths (p = 0.65, ANOVA,
n = 4). The response amplitude varies significantly
with stimulus pulse intensity (p = 0.0094, ANOVA,
n = 4) and width (p = 0.045, ANOVA, n = 4) for both
recording systems. The threshold for stimulus-evoked
CAPs was ≈0.4 V, and CAPs continued to increase
in amplitude with stimulus intensity until 1.5 V after
which a plateau was reached. An increasing trend was
observed in response to different pulse widths, but
with a slight decrease in CAP amplitude for pulses
greater than 100 µs. This variation in CAP amplitude
with stimulus intensity and width is independent of
the recording system (p = 0.29 and 0.26, respectively,
two-way ANOVA interaction term, n = 4).
The stimulus-response latency was nearly con-
stant across all stimulus pulse intensities (p = 0.72,
ANOVA, n = 4) and widths (p = 0.35, ANOVA, n = 4)
in both recording systems and there was no signific-
ant difference between recording systems (p = 0.23
for stimulus intensities, p = 0.13 for pulse widths,
ANOVA, n = 4, figures 6(f) and (g)).
3.5. Towards an MOA system
We fabricated patterns of miniaturised transducers
on substrates to construct MOAs (figures 7(a) and
(b)). The sensing interface is similar to electrodes in
MEAs, and the transducer element underlying each
sensing interface is analogous to amplifiers and fil-
ters underneath each pixel in active MEAs. A major
distinction is that the fabrication process we have
developed automatically scales the geometric area of
the transducer to match that of the sensing interface.
We compared the impedance properties of trans-
ducers and sensing interfaces at 1 kHz using elec-
trochemical impedance spectroscopy [31]. The inter-
face impedances were 9.08 kΩ and 0.58 kΩ, and the
transducer impedances were 6042.22 kΩ and 397.50
kΩ at sensing geometrical areas of diameters 250 µm
and 1 mm, respectively. The relative ratio of the sens-
ing transducer impedance to interface impedance was
665 vs. 685 (i.e. a 3% change for a 16-fold difference
in area).
We proceeded to demonstrate the feasibility of
reading out the signal from a MOA using a confocal
microscope system. With a line scan speed of 16 kHz
we were able to optically capture sinuosidal voltage
waveforms (figure 7(c)).
4. Discussion
There is a plethora of scientific literature on optrodes
as waveguides for delivery of light for optogenetic
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A Al Abed et al
manipulation or as fibre optic probes for voltage- or
calcium-sensitive imaging [32, 33]. In the commer-
cial neurotechnology sphere, the term ‘optrode’ has
become synonymous with the earlier neuromodu-
lation technique. Despite a recent review highlight-
ing the advantages of electro-optical sensors as a
potential solution to overcome current constraints on
the miniaturisation of multi-channel brain sensors
[5], a biopotential sensing optrode had been elusive.
Using optics to observe changes in cell refractive
index and birefringence during electrical excitation
[34] has failed to materialise as mainstream tech-
niques. Theoretical studies have explored the feas-
ibility of optical fibre refractometry [35] and sur-
face plasmon resonance has been reported for neural
recording in chamber [36] and fibre tip [37] configur-
ations, but several issues remain unaddressed, includ-
ing refractometry sensor characterisation, poor signal
resolution, and a means of calibrating the output-
ted signal to biopotentials. Here, we present a prac-
tical implementation and experimental validation
of a sensing fluorophore-free optrode with proper-
ties suitable for electrophysiology and brain-machine
interface applications based on photonic techniques.
To the best of the authors’ knowledge this is the first
report of a passive optrode for electrophysiology sens-
ing. This passivity is underpinned by two features (a)
the DHF-LC electro-optical transducer draws no elec-
trical power, and (b) signal detection does not require
voltage-sensitive fluorophores or any other form of
cell labelling.
Increasingly clinical and biomedical research sys-
tems employ optical fibres to transmit signals from
amplifiers/pre-amplifiers to data acquisition units.
Our approach employs light to detect and trans-
mit electrophysiology signals further upstream, right
from the biological interface. Hence our optrode
transducers pave the way to borrowing techniques
from the photonics and telecommunications indus-
tries to enable a step-increase in sensing bandwidth.
We validated the electrophysiology sensing cap-
ability of single channel optrode devices ex vivo and
in vivo. Our results demonstrate that optrodes based
on DHF-LCs can detect CAPs from the rabbit sci-
atic nerve as well as rabbit cardiac electrograms with
comparable waveforms, and can reproduce stimulus-
intensity and stimulus-duration response amplitude
and latency curves similar to those obtained via a con-
ventional bioamplifier system.
We fabricated MOAs by sandwiching DHF-LCs
between a top insulating substrate with through-vias
as the biological interface and a bottom transpar-
ent conductive substrate (figure 7). MOAs interface
with tissue in a similar manner to conventional MEAs
and can accommodate any contact material, design,
or geometry (flat, protruding), including conform-
al/soft designs. Our MOA fabrication protocol and
transducer design produce an automatic scaling of tis-
sue interface impedance with input impedance. An
J. Neural Eng. 0 (2022) xxxxxx A Al Abed et al

Figure 7. Multi-optrode arrays (MOAs). (A) Photo of a multi-optrode array with contact and electro-optical transduction regions
of diameters 40–1000 µm. Tracks have been added for characterisation purposes. Scale bar; 2 mm. (B) Photo of multi-optrode
array. Scale bar for main image 10 mm and for inset 250 µm. (C) Time-series line scan demonstrating changes of intensity of light
reflected along a 912 µm line across a transducer with applied 1 V 1 kHz sinuoisodial wave. (D) Illustration of a potential
microscope setup for fluorophore-free multi-channel imaging of biopotentials. (E) Schematic of proposed optical multiplexing
scheme. Signal detection can be achieved via an array of photodetectors or photodetector pixels in a camera. (F) Bragg grating
waveguide wavelength division multiplexing for multi-channel recording. PM, polarisation-maintaining; DAQ, data acquisition
system.

in-depth characterisation and analysis of this feature
is provided in a separate publication. This distinc-
tion confers MOAs a major advantage over active
MEAs. As the size of the electrodes in high-definition
CMOS MEAs is reduced, the area underneath each
electrode pixel is also reduced, constraining the size
and design of onchip amplifiers. As a result miniatur-
isation is traded-off with increased amplifier noise
and a reduced interface impedance to amplifier input
impedance ratio [4]. MOAs avoid this trade-off as the
electro-optical transduction is passive, and the ratio
of the interface to transducer (input) impedance is
maintained relatively constant independently of sens-
ing interface size. In terms of the spatial resolution
of MOAs two factors could be seen as limit setting.
Firstly the size of the light beam that will be pro-
jected onto the optrode’s mirror and reflected back.
The use of collimated light minimises scattering and
optical cross-talk between adjacent pixels across the
thickness of the LC. However beam sizes of 10 µm
or less are practically achievable. Secondly electrical
cross-talk between adjacent pixels. Pilot modelling
studies suggest that this limit of channel packing
is around 15–20 µm edge-to-edge. Interestingly, for
practical purposes the DHF-LC layer thickness does
not influence the minimal lateral resolution as the
limit imposed by this factor is determined by the
helix pitch of DHF-LC cells in the Smectic-C∗ phase

14
J. Neural Eng. 0 (2022) xxxxxx
(<1 µm). The layer simply needs to be thick enough
to avoid surface anchoring effects and thin enough
not to absorb an unacceptable amount of light.
The construction of optrode devices allows for
customisable electro-optical transduction properties.
The combination of the LC layer thickness, the DHF-
LC types, and their alignment relative to the polar-
isation angle of incident light could be optimised to
achieve higher sensitivity or bandwidth. The main
performance-determining difference between the two
optrodes devices tested extensively in this study is
the LC alignment layer, demonstrating that optrode
transducers can be fabricated to target particular elec-
trophysiological applications. Care needs to be taken
to eliminate combinations that could introduce sig-
nificant signal distortion. In terms of sensitivity, we
have shown using time-domain ensemble averaging
that I3 and D3 optrodes devices are capable of detect-
ing a 10 µV sinusoidal signal (supplementary figure
3). Hence the physics of the electro-optical trans-
duction allows for detection of single unit activity
and local field potentials. We expect future iterations
of the photodetection circuitry and optimisation of
optrode manufacture to reduce the noise floor and
increase the SNR to levels on par with conventional
bioamplifier systems. Towards this aim we have pro-
posed operating photodiodes under a non-reverse
biased zero mode that eliminates the background
when transducing light signals into voltages so that
only the AC component of the signal is sensed. This
enables higher first stage amplification gain without
photodectector saturation and hence a larger SNR
[38].
The ability of LCs to modulate light in response
to electrophysiological signals was first demonstrated
by coating frog hearts with LCs [39]. In contrast
to that pioneering study, our optrodes employ the
electro-optical transduction properties of LCs in an
encapsulated device. The conductive and insulating
material of the biological interface can be chosen
from the myriad of materials that have passed rig-
orous cytotoxicity testing and this interface layer
protects tissue from exposure to LCs or alignment
material and allows for a biocompatible and hermetic
encapsulation.
5. Conclusion
We foresee several future implementations of single
and multi-channel optrode devices in both biomed-
ical research laboratory and clinical settings.
A single channel optrode could offer a univer-
sal substitute for current application-specific electro-
physiology headstages. The insensitivity of optrode
functionality to contact impedance means that an
optrode device can be used as a headstage in many
application scenarios by connecting it to electrodes
of different cellular positions (intra, cell attached,
extra), material (glass pipettes, metal, polymer), and
15
A Al Abed et al
sizes/impedances (100 MΩ microelectrodes to 1 KΩ
skin electrodes). Immunity of the light signal to
electromagnetic interference confers an additional
advantage. A single optrode with adaptors to various
electrodes, with associated universal photodetection
and recording system, presents a cost effective solu-
tion for electrophysiology laboratories, instead of the
current practice of purchasing dedicated headstages
and amplifiers for each recording application.
MOAs will enable extracellular voltage imaging
in a fluorophore-free manner. In contrast to optical
voltage imaging, cells and tissue will not degrade from
dye-excitation associated photo-toxicity and dura-
tion of experiments will not be limited by the life-
time (photobleaching) of the fluorophores. MOAs
can also be integrated into existing inverted two-
photon, confocal or epifluoresence microscopy sys-
tems (figure 7(d)). Optical losses in the transducer
are small. Unconstrained by the need for longer
pixel dwell times to compensate for the low emis-
sion of fluorophores, the galvo or resonant scanners
or imaging cameras can be driven to their full poten-
tial to achieve significantly higher frame-rates while
maintaining good sensitivity. The tissue-interfaces of
MOAs could be fabricated to be either flat surfaces or
penetrating for extracting depth information, and in
both cases would not require fluourophores. Coupled
with microscopy systems, MOAs will enable high
channel count high density electrophysiology record-
ing without wires or interconnects associated with
CMOS MEAs.
Crucially, the potential advantages of our optrode
technology extend beyond the aforementioned labor-
atory settings to addressing clinical electrophysiology
monitoring challenges. The number of electrodes
in regulatory approved cardiac and neurology dia-
gnostic systems remains limited. The most common
intracardiac rhythm mapping catheters have a hand-
ful of channels, with ‘high-resolution’ variants offer-
ing 64 channels [40, 41]. Similarly intraoperative
cortical monitoring arrays have around twenty elec-
trodes. These systems are limited by challenges associ-
ated with wiring and interconnects required to trans-
mit signals captured by electrodes to monitoring sys-
tems. In addition, to maintain a good SNR, electrode
sizes are still relatively large.
Our optrode system offers an alternative that
can overcome these limitations. The early detec-
tion and transmission of electrophysiology signals
using photons rather than electrons should enable
the deployment of existing and mature optical tele-
communication technologies that will play a disrupt-
ive role in terms of packing density, channel count
and system-wide bandwidth. Indeed, the proposed
approach not only leverages optical fibres’ inherent
high-bandwidth (typically ≫1 Gb s−1 ) but is an ideal
example where polarisation and wavelength proper-
ties of light—properties not existing in the electrical
domain—can be used to radically increase packing
 J. Neural Eng. 0 (2022) xxxxxx
density by providing the required addressing and
multiplexing methodologies. Furthermore, interfero-
metric technologies such as (holographic) Bragg grat-
ings can be implemented on planar optical chips
(e.g. see figures 7(e) and (f)) to provided addressing
schemes based on wavelength that potentially could
support thousand of channels for the same optical
delivery catheter sheath size. This technology–known
as wavelength division multiplexing—already exists
in a polished form in the telecommunication industry
and can be readily adapted to our intended goals.
Data availability statement
The data that support the findings of this study are
Q8 available upon reasonable request from the authors.
Acknowledgments
The authors would like to thank Ms Naomi Craig for
her assistance in animal welfare, anaesthesia, and sur-
gery, Yuting Dou and Dr Ales Benda for conduct-
ing and guidance with the micorscopy time-series line
scans, and Emilie Revol for her pilot work on optrode
detection of cardiac electrograms. This work was per-
formed in part at the NSW Node of the Australian
National Fabrication Facility and the UNSW Biomed-
ical Imaging Facility.
This work was supported by the Australian
Research Council (Discovery Project Grant Numbers
DP160104625 and DP200102825), the Australian
National Health and Medical Research Council (Ideas
Project Grant Number 2002282), and the US Office of
Naval Research Global (Grant Number N62909-18-1-
2147).
Ethical statement
All experimental procedures were conducted in
accordance with the Australian Code for the Care
and Use of Animals for Scientific Purposes 8th Edi-
tion 2013, and were approved by the University of
New South Wales Animal Care and Ethics Commit-
tee (18/47B).
ORCID iDs
Amr Al Abed  https://orcid.org/0000-0002-5560-
4771
Yuan Wei  https://orcid.org/0000-0002-1346-9161
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