Article

Single-Neuron Correlates of Decision Confidence in

the Human Medial Temporal Lobe
Highlights Authors
d MTL neurons are modulated by confidence and reaction time Alexander Unruh-Pinheiro,
in decision making Michael R. Hill, Bernd Weber,
Jan Boström, Christian E. Elger,
d Independent subpopulations of neurons reflect confidence Florian Mormann
and reaction times

d Correlations lasted for as long as they were relevant for the

Correspondence
behavioral task florian.mormann@ukbonn.de

d Transient changes in neuronal activity were not explained by In Brief

value Unruh-Pinheiro et al. demonstrate both a
persistent and a transient modulation of
the neural activity in the human medial
temporal lobe (MTL) during decision
making. Persistent activity correlated
with decision confidence and reaction
time, but transient activity was not
explained by value, calling into question
the MTL’s role in valuation.

Unruh-Pinheiro et al., 2020, Current Biology 30, 1–11

December 21, 2020 ª 2020 Elsevier Inc.
https://doi.org/10.1016/j.cub.2020.09.021 ll
 Please cite this article in press as: Unruh-Pinheiro et al., Single-Neuron Correlates of Decision Confidence in the Human Medial Temporal Lobe, Cur-
rent Biology (2020), https://doi.org/10.1016/j.cub.2020.09.021

ll

Article
Single-Neuron Correlates of Decision Confidence
in the Human Medial Temporal Lobe
Alexander Unruh-Pinheiro,1 Michael R. Hill,1 Bernd Weber,1,2 Jan Boström,3 Christian E. Elger,1 and Florian Mormann1,4,*
1Department of Epileptology, University of Bonn Medical Center, Venusberg-Campus 1, 53127 Bonn, Germany
2Center for Economics and Neuroscience, University of Bonn, Venusberg-Campus 1, 53127 Bonn, Germany
3Department of Neurosurgery, University of Bonn Medical Center, Venusberg-Campus 1, 53127 Bonn, Germany
4Lead Contact

*Correspondence: florian.mormann@ukbonn.de
https://doi.org/10.1016/j.cub.2020.09.021

SUMMARY

The human medial temporal lobe (MTL) has been suggested to play a role in valuation. However, little is
known about its role in binary decisions and metacognition. We performed two decision-making tasks while
recording from neurons in the human MTL. During a break, subjects consumed their preferred food item to
satiation and subsequently repeated both tasks. We identified both a persistent and a transient modulation of
the neural activity. Two independent subpopulations of neurons showed a persistent correlation of their firing
rates with either decision confidence or reaction times. Importantly, the changes in confidence and reaction
time between experimental sets were accompanied by a correlated change in the neural activity, and this cor-
relation lasted as long as it was relevant for the behavioral task. Previous studies have suggested a transient
modulation of the neural activity in the human MTL correlated with subjective value. However, in our study,
neither subjective value nor unsigned value could explain this transient activity better than the nutritional fea-
tures of the stimuli, calling into question the role of the human MTL in valuation.

INTRODUCTION
In value-based decision making, subjective value can be defined
as a function that assigns a utility to items or actions. To compute
subjective value, we need to integrate different internal and
external sources of information, such as hunger or the different
available options, onto a common scale that allows comparing
different options [1]. Subjective value also plays a key role in rein-
forcement learning, behavioral economics, the free-energy prin-
ciple, and Bayesian brain models [2, 3]. Two previous human sin-
gle-unit studies have suggested the presence of neural
correlates of subjective value in the human medial temporal
lobe (MTL) [4, 5]. By definition, subjective value must integrate
both positive and negative rewards onto a common monotonic
unidimensional scale [1]. Thus, neural correlates of value should
be monotonically modulated and should span both the positive
and the negative reward domain. However, these studies only
tested positive values and therefore could not show whether
both valuation domains were integrated at the single-neuron
level, and they could not control for unsigned value. In the neuro-
economics literature, unsigned value is sometimes referred to as
salience [6], and it can be a statistical confound of subjective
value when testing only positive or only negative values. More-
over, subjective value is a dynamic property of the stimuli, i.e.,
because our preferences need not be stable over time and
context, a neuronal correlate of value must reflect these fluctua-
tions. This offers an experimental opportunity to dissociate value
from static properties of the stimuli, such as the content of salt or
sugar. These controls are important, because the amygdala has
been suggested to play a role in both unsigned value [7, 8] and
nutritional features [9]. Here, we wanted to dissociate subjective
value correlates from these potential confounds.
During binary decisions, subjective values of different items
may be compared to guide choice and may be integrated to
compute several key decision variables, such as the (unsigned)
difference in value or the chosen value. Moreover, during deci-
sion making, we constantly monitor the certainty of our deci-
sions, a metacognitive function termed decision confidence
[10]. Decision confidence has a strong influence on our deci-
sions: it is essential for learning, decision optimization, and
sequential decisions [11–13]. Previous studies have suggested
that decision confidence arises as a metacognitive readout of
the value difference [14]. Bayesian models have also proposed
that confidence could be defined as the probability that a deci-
sion is correct [15, 16]. In these models, confidence represents
a scalar summary statistic of the uncertainty of a decision, a
graded measure of belief that can be explicitly reported in an
experiment [13, 17].
Confidence, choice, and reaction time (RT) are three of the
main observables of a decision [18]. Drift-diffusion models of
evidence accumulation propose a unified mechanism for the
computation of these three variables [19]. Previous studies in
the macaque parietal cortex have suggested a unified neural
mechanism of evidence accumulation for the computation of
choice, RT, and confidence [18, 20, 21]. Independent correlates
of confidence, however, have also been observed in the frontal
cortex and subcortical areas [14, 22–28]. Interestingly, a previ-
ous study has reported confidence correlates in the human

Current Biology 30, 1–11, December 21, 2020 ª 2020 Elsevier Inc. 1
 Please cite this article in press as: Unruh-Pinheiro et al., Single-Neuron Correlates of Decision Confidence in the Human Medial Temporal Lobe, Cur-
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ll
Figure 1. Experimental Paradigm
Two cognitive tasks were performed. Top: in a valuation task, subjects were
presented with 10 salty and 10 sweet stimuli and provided a rating of sub-
jective value. Positive and negative ratings were allowed. Each stimulus was
presented 3 times, amounting to a total of 60 trials. Bottom: in a two-alter-
native, forced-choice task (2AFC), subjects were sequentially presented with a
pair of stimuli from the same 20 items: first, stimulus termed A was displayed
on the screen (presentation ‘‘A1’’), followed by stimulus B, and finally the first
stimulus, A, was presented a second time (presentation ‘‘A2’’). Subjects
selected their preferred stimulus and provided a rating of their subjective
confidence in the decision. During the response screen, the stimuli were
located either on the left or right in a balanced randomized manner. Each pair
of stimuli was presented once, resulting in a total of 190 trials. During a break
after the first set of both paradigms, the stimulus chosen most often in the
2AFC task was consumed to satiation. A second set of both tasks was carried
out after the break. Subjects were instructed they would consume their
preferred item after the second experimental set also, to control for motivation.
MTL during a memory task [29]. The MTL is important for deci-
sion making, and therefore, we wondered whether these confi-
dence correlates could be extended to the context of decision
making and whether these were (in)dependent of choice and
RT.
In this study, we assessed whether single-neuron correlates of
value, decision, or metacognitive variables arise in the human
MTL. We designed two tasks using food-item pictures as stimuli:
a valuation task and a two-alternative forced-choice task (2AFC).
Subjects performed both tasks twice, separated by a break, dur-
ing which the preferred item was consumed to satiation. By
repeating the task and perturbing the homeostatic state through
consumption, we were aiming to trigger a change of preferences
to dissociate static and dynamic variables. Moreover, subjects
reported both positive and negative values. Thus, we explicitly
provided a neutral value, the zero value that served as an objec-
tive reference to dissociate value and unsigned value. Finally,
subjects reported their confidence during each binary decision,
to test whether confidence correlates arise in the human MTL
during decision making.
2 Current Biology 30, 1–11, December 21, 2020
Article
RESULTS
Task and Behavioral Results
Twelve subjects performed a valuation task and a 2AFC task
(Figure 1). During a break after the first set of both tasks, the stim-
ulus chosen most often in the 2AFC task was consumed to sati-
ation, followed by a second experimental set of both tasks. Sub-
jects were instructed they would also consume the preferred
item from the second experimental set to control for motivation.
During each trial of the valuation task, subjects were instructed
to provide a value rating for 20 food items, including both positive
and negative values. Based on the literature, unsigned value
should be negatively correlated with RT [30]. Moreover, this
study suggested an asymmetric relationship of the unsigned
value with RT for objective learned values. We implemented a
mixed-effects linear regression predicting RT and including as
independent variables unsigned value, valence (i.e., a binary var-
iable for positive versus negative value), and an interaction term.
Unsigned value was significantly and negatively correlated with
the RT at the group level (unsigned value: b =
0.20; t1436 =

6.76; p < 10
10), but no asymmetry was found in the influence
of unsigned value between the positive and negative domain (un-
signed value 3 valence: b = 0.01; t1436 = 0.56; p = 0.5790). Thus,
the unsigned value was symmetrically correlated with RT for
both positive and negative subjective values.
During the 2AFC task, two stimuli A and B were sequentially
presented in the order A-B-A, and subjects were instructed to
choose their preferred item and to report their decision confi-
dence. At decision time, stimuli were alternatively displayed on
the left or right side of the screen. To compare the response
behavior of our subjects with previously reported behavioral re-
sults, we designed a mixed-effect linear model predicting confi-
dence [12] (Figure 2A). Significant predictors were unsigned dif-
ference in value (b = 0.46; t4556 = 9.73; p < 10
21), RT (b =
0.07;
t4556 =
3.53; p < 10
3), and summed value (b = 0.33; t4556 =
4.93; p < 10
6; formal model comparison in Figure S1A and Table
S1). Furthermore, we implemented a logistic mixed-effect model
predicting the probability of choosing stimulus A versus stimulus
B. We found significant predictors for the difference in value (b =
5.64; t4554 = 7.67; p < 10
13), the interaction between the differ-
ence in value and confidence (b = 2.87; t4554 = 6.41; p < 10
9),
and the interaction between the difference in value and the
summed value (b =
0.76; t4554 =
2.52; p = 0.012; Figure 2B).
We additionally split the data into the four levels of confidence
and performed a mixed model with difference in value as a pre-
dictor (Figure 2C). The slope of the difference in value as a pre-
dictor was monotonically steeper with higher confidence re-
ports, reflecting higher accuracy in the decision with higher
confidence reports. We repeated all these models of behavior
using log-transformed RT, yielding very similar results. Together,
these analyses replicated well the results from Folke et al. [12]
and indicated that reporting confidence at the time of the deci-
sion, instead of in a second, separate report as usually per-
formed in the literature, led to qualitatively identical behavior.
Subjects performed two consecutive experimental sets of
tasks separated by a break. We expected that the behavioral
variables would fluctuate between experimental sets, e.g., due
to preference drift, task repetition, or noise in the computation
of decision variables. To further increase the chances of finding
 Please cite this article in press as: Unruh-Pinheiro et al., Single-Neuron Correlates of Decision Confidence in the Human Medial Temporal Lobe, Cur-
rent Biology (2020), https://doi.org/10.1016/j.cub.2020.09.021
Article
a preference change, subjects consumed their preferred item to
satiation. To quantify these fluctuations, we calculated Spear-
man’s rank correlation for each variable between the first exper-
imental set of tasks, before the satiation break, and the second
set. The average of Spearman’s rho across subjects showed
that confidence (r = 0.35 ± 0.31 SD), RT during 2AFC task (r =
0.12 ± 0.13 SD), RT during valuation task (r =
0.01 ± 0.27
SD), and unsigned value (r = 0.40 ± 0.20 SD) were perturbed
and partially decorrelated. Fluctuations of subjective value, how-
ever, were lower in comparison. This was true both when value
was estimated as the mean of the value ratings during the valu-
ation task (r = 0.63 ± 0.29 SD) as well as when estimated based
on value rankings, i.e., the number of times that each stimulus
was chosen in the 2AFC task (r = 0.67 ± 0.35 SD; Figure S1B).
However, both the value of the consumed item as well as the
value of the items sharing the consumed taste (sweet versus
salty) were significantly devaluated on average across subjects
(Table S2). This was not the case for the items sharing the non-
consumed taste, showing that the satiation manipulation indeed
had an effect on valuation.
Decision Confidence Is Correlated with Sustained
Changes in Firing Rate
We recorded from 830 units (480 single units and 350 multi-
units) using microelectrodes implanted bilaterally in the amyg-
dala (A) (265 units), hippocampus (H) (305 units), entorhinal cor-
tex (EC) (152 units), and parahippocampal cortex (PHC) (108
units). In all analyses, both single-unit and multi-unit activities
ll
Figure 2. Behavioral Results
(A) Fixed-effect coefficients of a linear mixed-effect
model of confidence with predictors: unsigned
difference in mean value rating (|DV|); reaction time
(RT); and summed mean value rating of the stimuli
(sum).
(B) Fixed-effect coefficients of a logistic multi-level
model predicting the probability of choosing the
stimulus A as a function of the difference in mean
value rating (DV), confidence, sum, and the inter-
action term between DV and confidence, as well as
the interaction term between DV and sum.
(C) Logistic mixed-effect model showing the
probability of choosing the stimulus A as a function
of the mean value rating difference between stimuli
A and B. Data were split into four models, one for
each of the four possible confidence steps on the
Likert scale. Error bars represent 95% confidence
intervals. *p < 0.05; **p < 0.01; ***p < 0.001; two-
sided t test. See also Figure S1 and Table S1.
were conjointly analyzed, as is typical
for human unit recordings.
We used a sliding-window Spearman’s
rank correlation (stepsize = 10 ms) to test
whether the firing rates monotonically
correlated with cognitive variables. For a
neuron to be significant, correlations had
to be simultaneously significant in both
experimental sets of tasks and with the
same correlation direction. We found
units significantly correlated with decision confidence (Fig-
ure 3A). The activity of these units symmetrically correlated
with subjective confidence, regardless of the temporal and
spatial location of the stimuli (Figure 3B). At the population level,
the significance of the fraction of significant units was assessed
using a binomial test, and cluster-based permutation tests were
performed to correct for multiple comparisons over time. A sig-
nificant fraction of units correlated with confidence for a sus-
tained period of time (Figure 4A), and this significant correlation
stopped abruptly after confidence was reported. Because we
also observed a transient stimulus-locked activity (see below),
we aimed to avoid an overlap between both signals. We defined
our analysis window from the Likert-scale presentation until
2,350 ms later, which matched the shortest inter-trial duration
across all subjects, ensuring no overlap with the subsequent
trial. The mean proportion of units correlated with confidence,
averaged across the steps included in the analysis window,
was 11.81% (p < 10
14; one-tailed binomial test). Regional anal-
ysis showed that A, H, and EC contained highly significant pro-
portions (A: 11.32%, p < 10
4; H: 11.15%, p < 10
4; EC:
15.13%, p < 10
5), whereas PHC barely reached significance
(9.26%; p = 0.0443; Figure S2B). As a control, we found no evi-
dence for asymmetric correlation with confidence (signed confi-
dence based on the temporal presentation [5.54%; p = 0.2574];
signed confidence based on the spatial location [5.30%; p =
0.3670]). Similarly, binary choices were not significant, neither
based on the temporal presentation (5.78%; p = 0.1690) nor on
the spatial location (5.18%; p = 0.4276).
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ll
Confidence reports per trial varied between the two sets of
experiments. We tested whether these changes were accom-
panied by correlated changes in the neural activity. We
compared for each unit the magnitude of the Spearman corre-
lation coefficient between the actual confidence ratings and
the firing rates, averaged across both experimental sets,
with a surrogate version for which we swapped ratings be-
tween the experimental sets, meaning that firing rates from
one set were now correlated with confidence ratings of the
corresponding trials from the other set (see STAR Methods
for details). If the change in confidence was accompanied
by a correlated change in the neural activity, the magnitude
of the actual correlation should explain the variance better
than the swapped surrogate. We used a one-tailed Wilcoxon
signed-rank test to assess whether the magnitude of the
actual correlations was larger than their swapped surrogate
version. We performed this analysis including all neurons
and repeated it for the subset of neurons correlated with
either confidence or its swapped surrogate. We found that
the signed-rank tests were consistently significant for a sus-
tained period of time (a = 0.05; Figure 4A, bottom). As an alter-
native analysis, we analyzed for each neuron whether the
change in firing rate between experimental sets correlated
with the change in confidence on a trial-by-trial basis, again
with a sliding-window Spearman’s correlation. The average
across the steps in the analysis window yielded 8.31% signif-
icantly correlated neurons (p < 10
4; one-tailed binomial test).
These results suggest that neurons in the human MTL were
correlated with decision confidence and updated their state
on the set-by-set basis. Importantly, the cause of the change
4 Current Biology 30, 1–11, December 21, 2020
Article
Figure 3. Single Unit Correlated with Confi-
dence
(A) Located in the right EC, this unit showed a
monotonically increasing firing rate positively
correlated with the subjective confidence ratings.
Left: neuronal activity aligned to the current trial
onset is shown. Right: neuronal activity aligned to
the subsequent trial onset is shown. Top: black
horizontal bars indicate the analysis windows
during which this unit was significantly correlated
with confidence simultaneously in both experi-
mental sets and with the same sign of the corre-
lation. Middle: raster plot of action potentials is
shown. Bottom: mean instantaneous firing rates
for different confidence levels are shown
(Gaussian kernel s = 200 ms; stepsize = 10 ms).
Violet, orange, yellow, and blue lines represent the
different possible values of confidence on the
Likert scale, from 25 to 100. Shaded areas repre-
sent the SEM. Light gray vertical bars indicate the
presentation of stimulus A; the dark gray vertical
bar in between indicates presentation of stimulus
B. Inset shows the density plot of all spikes.
(B) Mean firing rate averaged across the window of
interest. Left: confidence reports for different
spatial locations of the chosen stimulus during the
response time, i.e., left versus right, did not affect
the correlation with confidence. Right: the activity
was also independent of the temporal location of
the chosen stimulus, i.e., stimulus A versus B. Error
bars represent SEM.
in the cognitive variable is of little importance, as long as the
change in the cognitive variable is accompanied by correlated
changes in neural activity.
RT Correlates with the Activity of Single Neurons
Surprisingly, we also found a significant proportion of neurons
correlated with RT (13.01%; p < 10
15). The analysis of RT was
initially planned as a control. Interestingly, the neural correlate
of RT started to drop right after the binary choice was made,
whereas the neural correlate of confidence persisted until the
confidence report (Figure 5). Regional analysis showed a signif-
icant effect in the four regions (A: 12.08%, p < 10
5; H: 13.44%,
p < 10
7; EC: 13.82%, p < 10
4; PHC: 11.11%, p = 0.0080; Fig-
ure S2B). We then repeated the ‘‘actual versus swapped’’
signed-rank test described above to assess whether the
changes in RT were reflected by the change in the neuronal ac-
tivity. We found a sustained significant effect (Figure 4B, bot-
tom). Likewise, the proportion of neurons whose change in firing
rate across sets significantly correlated with the change in RT
was highly significant (10.12%; p < 10
8). We additionally tested
whether neural correlates of RT and confidence arose during the
valuation task, finding no evidence for a persistent modulation of
the neural activity (Figure S3). Moreover, because subjects
answered by repeatedly pressing a keyboard, motor signals
arose as a potential confound of confidence. We designed
both the valuation and decision task with the same answering
methodology. Because no evidence of a persistent modulation
of the neural activity was found during the valuation task, motor
signals are an unlikely explanation of the neural activity
(Figure S3).
 Please cite this article in press as: Unruh-Pinheiro et al., Single-Neuron Correlates of Decision Confidence in the Human Medial Temporal Lobe, Cur-
rent Biology (2020), https://doi.org/10.1016/j.cub.2020.09.021
Article
analysis (all, n = 830), which was repeated for the subset of significantly correlated units (selected, n changes at each step). The black horizontal lines above and
below the p values indicate the significant clusters in the cluster permutation test (p < 0.01).
(B) Same analyses for reaction time (RT). See also Figures S2–S5 and Tables S3 and S4.
Confidence and RT Are Correlated with Two Different
Neuronal Subpopulations
Due to the similarity of both variables (magnitude of the correla-
tion between confidence and RT, averaged across subjects: r =
0.31 ± 0.13 SD), we wondered whether only one of these vari-
ables could explain most of the variance of the persistent activ-
ity. If not, we wanted to know whether different subpopulations
were correlated with the two variables or whether a single pop-
ulation of neurons was responsible for both correlates. We per-
formed three analyses to test this.
First, we tested which of the variables could better explain the
persistent activity. We computed the Spearman’s rho in the
above-mentioned sliding window. Because there was a partial
overlap between the cell identities, we performed a model selec-
tion on the ambivalent units: if a cell was significantly correlated
with the two variables at the same time, we assigned this ambiv-
alent unit to the variable with the larger magnitude of rho. Thus,
each cell at each time bin was only included in one of the popu-
lations, to avoid double counting of the units. After this proced-
ure, both signals were still highly significant (confidence:
10.12%, p < 10
8; RT: 11.81%, p < 10
14; Figure S4A). Next,
we performed a post hoc binomial test on these proportions:
assuming a chance level of 0.5 (no preference), we tested
whether the fraction of neurons for RT was higher than the frac-
tion for confidence. We found no significant difference between
the fraction of neurons (p = 0.1676; one-tailed binomial test).
Thus, neither of the variables was a statistically better model of
the persistent activity.
Second, we tested whether different subpopulations corre-
lated with these variables. Here, one of the variables was
ll
Figure 4. Population Results of the Persis-
tent Activity
Left: neuronal activity aligned to the current trial
onset is shown. Right: neuronal activity aligned to
the subsequent trial onset is shown. Light gray
vertical bars indicate the presentation of stimulus
A; the dark gray vertical bar in between indicates
presentation of stimulus B.
(A) Top: proportion of neurons significantly corre-
lated with confidence, pooled across regions (n =
830). The instantaneous firing rate of each cell was
rank correlated with the trial-by-trial confidence
report in a sliding-window correlation (Gaussian
kernel s = 200 ms; stepsize = 10 ms). The alpha
level of significantly correlated neurons was set to
5% (black dashed line), and the significance of
their proportion was assessed using a one-tailed
binomial test. The red dashed line shows the pro-
portion of cells that represent a p value of 0.001,
and all higher proportions were considered signif-
icant. A cluster-based permutation test was per-
formed to correct for multiple comparisons. The
solid curves indicate significant clusters (p < 0.01).
Bottom: in this analysis, we assessed whether the
change of confidence between experimental sets
was accompanied by a correlated change in the
neural activity (see text). p values of a Wilcoxon
signed-rank test over time are shown color coded
(steps of 10 ms). All cells were included in the top
treated as a control variable (e.g., confidence), and all the
units significantly correlated with this control were removed
from the analysis. For the remaining units, we calculated the
proportion of units significantly correlated with the other vari-
able (e.g., RT). Importantly, we did not remove the units from
the denominator of the proportion, to maintain a common
reference in the denominator and ensure comparability be-
tween variables. This yielded conservatively lower proportions
and thus higher p values. We then repeated the same analysis
for the other variable. If RT were enough to fully explain the
activity, we would expect a strong decrease in the proportion
of cells correlated with confidence and vice versa. Both the
proportion of units significantly correlated with RT and with
confidence remained highly significant (confidence: 8.80%,
p < 10
5; RT: 10.00%, p < 10
8; Figure S4B), suggesting
that two different subpopulations were responsible for the
two neural correlates.
Finally, we performed a rank-transformed multivariable
regression with RT and confidence as predictors of the firing
rate of each unit, to test the independence of the variables
when statistically controlled at the unit level. The mean fraction
of units correlated with the unique variance of both variables re-
mained highly significant (confidence: 10.12%, p < 10
8; RT =
12.53%, p < 10
16; Figures S4C and S4D), suggesting indepen-
dence of both signals. Taken together, these analyses suggest
that the neural correlates of confidence and RT in the MTL are in-
dependent and represented by different subpopulations of neu-
rons. Note that neurons showed a symmetric distribution of pos-
itive and negative correlation coefficients for RT and confidence
(Figure S5).
Current Biology 30, 1–11, December 21, 2020 5
 Please cite this article in press as: Unruh-Pinheiro et al., Single-Neuron Correlates of Decision Confidence in the Human Medial Temporal Lobe, Cur-
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ll
values of a Wilcoxon signed-rank test over time are shown (steps of 10 ms). All cells were included in the top analysis (all, n = 830), which was repeated for the
subset of significantly correlated units (selected, n changes at each step). The black horizontal lines above and below the p values indicate the significant clusters
in the cluster permutation test (p < 0.01).
(B and D) Same analyses for reaction time (RT).
Confidence and RT Explain the Variance in Neuronal
Activity Better Than Any Other Variable
Because numerous decision variables are mechanistically
coupled and statistically dependent, we next performed a quan-
titative analysis to compare confidence and RT against other de-
cision variables and stimulus-related variables, such as the cho-
sen value, the subjective value, and nutritional or visual features.
A total of 122 variables were compared as alternative explana-
tions. First, a ranking analysis was performed. We computed
the mean proportion of neurons significantly correlated with
each variable, averaged across the time steps of the analysis
window mentioned above, and sorted the variables by their
mean proportion. Confidence and RT showed the highest
mean proportion of neurons among all variables (RT: 13.01%;
confidence: 11.81%). Importantly, confidence and RT were
also the leading two variables in each individual region
(Figure S2A).
To quantify the significance of confidence and RT as the best
models, we applied the first analysis from the previous section to
each pair of variables, i.e., we sorted the ambivalent units by the
magnitude of the correlation and subsequently performed the
post hoc binomial test. As previously noted, RT proved slightly
better than confidence, but this difference was not significant
(p = 0.1676). Notably, both confidence and RT were significantly
better than all other tested variables (all post hoc tests p < 0.05;
Table S3). The unsigned difference in value and chosen value
were the third and fourth best models. We repeated the above-
mentioned subpopulation and multivariable regression analyses
to compare confidence and RT with the chosen value and the
6 Current Biology 30, 1–11, December 21, 2020
Article
Figure 5. Persistent Activity Aligned to
Choice Response and Confidence
Response
Light gray vertical bars indicate the presentation of
stimulus A; the dark gray vertical bar indicates
presentation of stimulus B.
(A and B) Neural activity aligned to binary choice
response.
(C and D) Neural activity aligned to confidence
response.
(A and C) Proportion of neurons significantly
correlated with confidence, pooled across regions
(n = 830). The instantaneous firing rate of each cell
was rank correlated with the trial-by-trial confi-
dence report in a sliding-window correlation
(Gaussian kernel s = 200 ms; stepsize = 10 ms).
The alpha level of significantly correlated neurons
was set to 5% (black dashed line), and the signif-
icance of their proportion was assessed using a
one-tailed binomial test. The red dashed line
shows the proportion of cells that represent a p
value of 0.001, and all higher proportions were
considered significant. A cluster-based permuta-
tion test was performed to correct for multiple
comparisons. The solid curves indicate significant
clusters (p < 0.01). Bottom: in this analysis, we
assessed whether the change of confidence be-
tween experimental sets was accompanied by a
correlated change in the neural activity (see text). p
unsigned difference in value (Table S4). These confirmed that
confidence and RT were better models of the persistent activity.
Neuronal Activity Is Correlated with Subjective Value
and Unsigned Value but Does Not Reflect the Change in
Preference between Experimental Sets
We also found a transient modulation of the activity time locked
to the stimulus presentations in both the valuation and the 2AFC
tasks. We quantified this neural activity as the mean proportion
of correlated neurons between the onset and offset of each stim-
ulus, averaged across the time steps of the sliding window, and
subsequently averaged across each of the four stimulus presen-
tations (valuation, A1, B, and A2). We found a significant propor-
tion of units whose firing rates were significantly correlated with
value ratings (9.88%; p < 10
8; Figure 6A) and value rankings
(9.88%; p < 10
8; Figure S2A). We also found a weaker but sig-
nificant correlation of the neural activity with the unsigned value
(8.55%; p < 10
4; Figure 6B). Regional analysis showed that A
and H were the main contributors to this transient activity
(Figure S2A).
Next, we analyzed whether the change in value or unsigned
value between experimental sets was reflected in the neuronal
activity. We repeated the ‘‘actual versus swapped’’ signed-
rank test described above. Surprisingly, this was not significant,
not even when performed on the subset of neurons significantly
correlated with value or unsigned value, respectively (Figures 6A
and 6B, bottom). We also tested the proportion of neurons
whose change in firing rate between experimental sets signifi-
cantly correlated with the change in variable and averaged
 Please cite this article in press as: Unruh-Pinheiro et al., Single-Neuron Correlates of Decision Confidence in the Human Medial Temporal Lobe, Cur-
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Article

Figure 6. Population Results of the Transient Activity

Left: valuation task; light gray vertical bar indicates stimulus presentation. Middle: 2AFC task, stimulus A variable (e.g., unsigned value of stimulus ‘‘A’’) is shown.
Right: 2AFC task, stimulus B variable (e.g., value of stimulus ‘‘B’’) is shown. Light gray vertical bars indicate the presentation of stimulus A; the dark gray vertical
bar in between indicates presentation of stimulus B.
(A) Top: proportion of neurons significantly correlated with mean value rating, pooled across regions (n = 830). The trial-by-trial instantaneous firing rate of each
cell was rank correlated with the mean value rating of each stimulus in a sliding-window correlation (Gaussian kernel s = 200 ms; stepsize = 10 ms). The alpha level
of significantly correlated neurons was set to 5% (black dashed line), and the significance of their proportion was assessed using a one-tailed binomial test. The
red dashed line shows the proportion of cells that represent a p value of 0.001, and all higher proportions were considered significant. A cluster-based per-
mutation test was performed to correct for multiple comparisons. The solid curves indicate significant clusters (p < 0.01). Bottom: in this analysis, we assessed
whether the change of value ratings between experimental sets was accompanied by a correlated change in the neural activity (see text). p values of a Wilcoxon
signed-rank test over time are shown (steps of 10 ms). All cells were included in the top analysis (all, n = 830), which was repeated for a subset of selected cells
(selected, n changes at each step). The black horizontal lines above and below the p values indicate the significant clusters in the cluster permutation test (p <
0.01).
(B) Same analyses for unsigned value.
(C) Proportion of neurons significantly correlated with taste, defined as a binary variable for salty versus sweet category.
See also Figure S2 and Tables S2 and S3.

Current Biology 30, 1–11, December 21, 2020 7

 Please cite this article in press as: Unruh-Pinheiro et al., Single-Neuron Correlates of Decision Confidence in the Human Medial Temporal Lobe, Cur-
rent Biology (2020), https://doi.org/10.1016/j.cub.2020.09.021
ll
across the analysis window. These were again not significant
(ranking: 6.14%, p = 0.0791; rating: 6.14%, p = 0.0791; unsigned
value: 5.54%, p = 0.2574). Therefore, we found no evidence that
the changes of these variables between experimental sets were
accompanied by correlated changes in the neural activity.
Value and Unsigned Value Were No Better Models Than
Nutritional Features of the Stimuli
We also performed a quantitative analysis of the transient stim-
ulus-locked activity. Here, we compared value and unsigned
value with other stimulus-dependent variables, such as nutri-
tional and visual features of the items, amounting to a total of
18 variables. In a ranking analysis, we found that taste, defined
as a binary variable for salty versus sweet, showed the highest
mean proportion of correlated units across stimulus presenta-
tions (9.88%; p < 10
8; Figure 6C), together with subjective value
(value ratings: 9.88%, p < 10
8; value ranking: 9.88%, p < 10
8).
These were closely followed by the content of sugar (9.28%; p <
10
6), salt (9.16%; p < 10
6), and fiber (9.04%; p < 10
6). Un-
signed value only reached the seventh position in the ranking
analysis (8.55%; p < 10
4; Figure S2A). Regional analysis
showed that, in the amygdala, both taste (12.45%; p < 10
5)
and fiber (11.32%; p < 10
4) were better than subjective value
(value ratings: 10.98%, p < 10
4; value ranking: 10.98%, p <
10
4). Subjective value was the leading variable in the hippo-
campus and EC and not significant in PHC (Figure S2A).
Moreover, our post hoc binomial test showed that subjective
value was not significantly better than taste (value ratings versus
taste: p = 0.4656), sugar (p = 0.2168), salt (p = 0.1692), or fiber
content (p = 0.1948; Table S3). Some nutritional features showed
a correlational similarity with subjective value, defined as the
magnitude of the correlation between both variables, averaged
across experimental sets and subjects (taste: r = 0.42 ± 0.28
SD; salt: r = 0.36 ± 0.27 SD; fiber: r = 0.36 ± 0.19 SD; sugar:
r = 0.40 ± 0.25 SD). Nevertheless, this only showed a partial
overlap, which is not sufficient to fully explain our findings in
terms of mere collinearity because, for instance, taste was as
good as subjective value in the MTL, and in the amygdala, taste
was even better than value.
DISCUSSION
In this study, we found neurons in the human MTL whose activity
was persistently correlated with decision confidence. Impor-
tantly, the change in confidence between experimental sets
was accompanied by a correlated change in the neural activity,
providing further evidence. Rutishauser et al. [29] have previ-
ously reported neuronal activity in the human amygdala and
hippocampus to be correlated with confidence. However, the
authors found two populations of memory- and novelty-
responding neurons asymmetrically correlated with confidence
reports, depending on their response type. Thus, these cells
could possibly have reflected memory strength rather than
confidence. Instead, we here found a spatially and temporally
symmetric neural correlate of confidence, independent of
choice. To our surprise, a different subpopulation of neurons
was persistently correlated with RT. Notably, both RT and confi-
dence better explained the variance of the neural activity than
other decision variables, such as the unsigned difference in value
8 Current Biology 30, 1–11, December 21, 2020
Article
or the chosen value, variables previously suggested to be
involved in confidence computation [15]. RT and confidence
also explained the persistent activity significantly better than
122 alternative variables, suggesting that these variables were
directly modulating the neural activity, instead of other lower-
level stimulus properties or decision variables. Moreover, the
persistent neural correlates of both RT and confidence were
only maintained as long as they were behaviorally relevant for
the task. Whereas the neural correlate of RT was maintained until
the binary choice was made, the correlate of confidence per-
sisted after the decision, until confidence was reported. Similar
graded persistent activity has previously been suggested by
attractor models of working memory in decision making [31, 32].
Previous studies have reported choice-correlated cells in the
primate amygdala [33, 34]. However, we found no evidence of
choice-related activity in our recorded neurons. Moreover,
several studies have previously described behavioral and neural
dissociation between confidence, RT, and choice [21, 28, 35–
37]. Here, we observed two independent subpopulations of neu-
rons correlated with either confidence or RT. Contrary to neurons
in the macaque parietal cortex [20], which show an accumulation
of evidence and at the same time represent choice, confidence,
and RT, neurons in the human MTL show a fragmented modula-
tion of the neural activity by decision observables. These results
suggest that confidence is computed somewhere upstream and
conveyed to the human MTL. On the other hand, the interpreta-
tion of a population of RT-correlated neurons is not trivial. The
task dependence of RT correlation suggests that RT might be
seen as a proxy for a non-observable or non-reported decision
variable, such as subjective decision difficulty. However, the
meaning of the activity of these cells can only be speculatively
addressed.
It has previously been suggested that, during valuation, valua-
tion confidence arises as a quadratic expansion of subjective
value [22]. We found no evidence of sustained activity correlation
with either valuation confidence or RT during the valuation task,
contrary to the 2AFC task. Two hypotheses could explain this dif-
ference: either confidence is correlated with the neural activity
specifically in the context of binary decisions or explicit report of
confidence is required for the neural correlate to emerge. Notably,
both behavioral tasks were presented with the same Likert scale
and same response methodology. Thus, the neural correlates of
confidence and RT are unlikely to reflect motor signals.
The amygdala has long been considered to play an essential
role in valuation [38–44]. Additionally, several animal studies
have proposed that the hippocampus and EC are also involved
and even necessary for reward processing [45–50]. More evi-
dence of the role of the amygdala in valuation has arisen from
its strong connection with the orbitofrontal cortex (OFC), tradi-
tionally acknowledged as the main neural locus for subjective
value [6, 51–56]. In a previous study from our group, we found
that the firing rates of single units correlate with subjective value
in the amygdala, hippocampus, and EC [5]. However, to our
knowledge, no study has provided evidence that neurons in
the human MTL reflect the changes in value. Here, we provide re-
sults confirming the existence of a task-invariant transient activ-
ity in the human MTL during visual stimulus presentation [5].
However, we could not confirm that subjective value is the
best explanation of this neural activity.
 Please cite this article in press as: Unruh-Pinheiro et al., Single-Neuron Correlates of Decision Confidence in the Human Medial Temporal Lobe, Cur-
rent Biology (2020), https://doi.org/10.1016/j.cub.2020.09.021

ll
Article

We found no evidence that the changes in subjective value be- d KEY RESOURCES TABLE
tween experimental sets were accompanied by a correlated d RESOURCE AVAILABILITY
change in the neural activity, a necessary requirement for a true B Lead Contact
value correlate. A limitation of this study is that the subjective values B Materials Availability
were not as strongly perturbed between experimental sets as other B Data and Code Availability
variables, which limits the sensitivity of our analysis. However, we d EXPERIMENTAL MODEL AND SUBJECT DETAILS
also found that the transient activity could be equally well explained B Subjects and implantation schemes
by the nutritional features of the stimuli. Importantly, these nutri- d METHOD DETAILS
tional features were only partially correlated with value. Thus, B Electrophysiological setup and data preprocessing
mere collinearity cannot fully explain these results. Wang et al. [9] B Cognitive paradigm
have shown that the projections of taste cortex to the amygdala d QUANTIFICATION AND STATISTICAL ANALYSIS
are necessary to link value and taste. Moreover, a neural correlate B Mixed-effect hierarchical models
of the subjective content of different nutrients has been reported in B Responses were not biased by the design.
human lateral OFC [57], and the amygdala is strongly connected to B Similarity and change of cognitive variables
OFC. An alternative explanation of the transient response could be B Firing rate estimation and windows of analysis
that amygdala activity correlates with lower level nutritional fea- B Identification of relevant variables
tures and this information would then be conveyed to the lateral B Estimates of subjective value were consistent
OFC and subsequently to the medial OFC, where it would be inte- B Variable update analysis
grated as an abstract and updated subjective value. B Quantitative comparison of variables
We also assessed unsigned value, which has previously been B Subpopulation and independence analyses
suggested to be related to the amygdala [7, 58]. Unsigned value B Changes of mind
was not even among the best candidates to explain the transient
signal, and its change between sets was not accompanied by a SUPPLEMENTAL INFORMATION
correlated change in firing rates. Note that the changes in un-
signed value of the stimuli were similar in size to those in decision Supplemental Information can be found online at https://doi.org/10.1016/j.
confidence. Thus, our results regarding the lack of correlated cub.2020.09.021.
change in the firing rates between experimental sets are more
conclusive than those for subjective value. Therefore, we here ACKNOWLEDGMENTS
found no evidence for a correlate of unsigned value in the activity
We thank all patients for their participation and Gert Dehnen, Marcel Bausch,
of the recorded units, which agrees with previous neuroimaging
Iria Dominguez, and Wolfram Schultz for discussion. This research was sup-
studies investigating unsigned value [30, 59]. However, because ported by the Volkswagen Foundation and the German Research Council
we recorded only a small fraction of MTL units, neurons in other (MO930/4-2, MO930/8-1 and SFB 1089). A.U.-P. was supported by the Fun-
subregions of the amygdala and the human MTL could reveal  de la Maza and the Obra Social La Caixa.
dación Barrie
value or unsigned-value correlates.
A limitation of our statistical approach is that it neglects the AUTHOR CONTRIBUTIONS
strength of the association between cognitive variables and the
activity of single neurons. In our analysis, we focused on whether F.M. and M.R.H. designed the study. C.E.E. recruited the patients. J.B. and
the fraction of neurons significantly correlated with a cognitive F.M. implanted the electrodes. M.R.H., A.U.-P., and F.M. collected the data.
variable was higher than expected by chance. Nonetheless, A.U.-P. conceptualized and performed the data analysis. A.U.-P. and F.M.
wrote the manuscript. B.W. provided the images of the stimuli. All authors dis-
there could be small subpopulations of neurons whose activity
cussed the results and commented on the manuscript.
was strongly correlated with other cognitive variables but whose
proportion of neurons did not reach significance after correction
DECLARATION OF INTERESTS
for multiple comparisons. However, correction for multiple com-
parisons is necessary to reduce the risk of false positives. The authors declare no competing interests.
In conclusion, our results show evidence for two segregated
subpopulations of neurons in the human MTL whose activity is Received: February 25, 2020
correlated with conscious reports of confidence and with RT dur- Revised: July 17, 2020
ing decision making. Both variables showed a persistent correla- Accepted: September 7, 2020
Published: October 8, 2020
tion with the neural activity as long as these variables were relevant
for the task, and the change in neural activity between experi-
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STAR+METHODS

KEY RESOURCES TABLE

REAGENT or RESOURCE SOURCE IDENTIFIER

Software and Algorithms
MATLAB MathWorks https://www.mathworks.com
Custom-Built This paper https://github.com/AlexanderUnruh/
Code and Data Single-Neuron-Correlates-of-Decision-
Confidence-in-the-Human-Temporal-Lobe.git
Combinato [60] https://github.com/jniediek/combinato/
Psychtoolbox https://psychtoolbox.org

RESOURCE AVAILABILITY

Lead Contact
Further information and requests for resources should be directed to and will be fulfilled by the Lead Contact, Florian Mormann
(florian.mormann@ukbonn.de).

Materials Availability
This study did not generate new unique reagents.

Data and Code Availability

Data and custom-built MATLAB code can be found in a GitHub repository (https://github.com/AlexanderUnruh/
Single-Neuron-Correlates-of-Decision-Confidence-in-the-Human-Medial-Temporal-Lobe.git).

EXPERIMENTAL MODEL AND SUBJECT DETAILS

Subjects and implantation schemes

Twelve patients (6 female and 6 male, all right-handed, mean age 40 ± 13.62 SD, range: 20-62) with pharmacologically intractable
temporal lobe epilepsy were implanted with intracranial electrodes for clinical evaluation of possible surgical removal of the epileptic
focus. Left-hemispheric seizure focus was confirmed in 7 patients, right-hemispheric focus in 4 patients, and a bilateral focus was
present in 1 patient. Electrode locations was based exclusively on clinical criteria and verified by computed tomography (CT) and
magnetic resonance imaging (MRI). Implanted regions included: amygdala (A: 12/12 patients), hippocampus (H: 11/12), parahippo-
campal cortex (PHC: 11/12) and entorhinal cortex (EC: 10/12). All regions were implanted bilaterally. All patients provided written,
informed consent for the surgical implantation of the electrodes and for participating in the cognitive experiment. All experiments
were approved by the Medical Institutional Review Board of the University of Bonn.

METHOD DETAILS

Electrophysiological setup and data preprocessing

Patients were implanted with Behnke-Fried Depth Electrodes (Ad-Tech, Racine, WI). Nine microwires of 40 mm diameter protruded
from the tip of each electrode: eight high-impedance recording wires and one low-impedance reference wire. Electrodes were con-
nected to a headstage pre-amplifier (CHET-10) which in turn was connected to an ATLAS data acquisition system (Neuralynx, Boze-
man, MT) and processed with Cheetah software. The signal was band-pass filtered between 0.1 and 9000 Hz and recorded at a sam-
pling frequency of 32768 Hz. All units were semi-automatically sorted using Combinato [60]. If more than one cluster was detected on
a given microwire, manual classification of the clusters was performed in order to ensure that the selected units were sufficiently
different based on the spike shape, inter-spike interval distribution, and cross-correlation of the spike-times between clusters.
Finally, each identified unit was manually classified as single- or multi-unit based on the spike shape, inter-spike interval distribution
and the presence of a refractory period.

Cognitive paradigm
Patients performed cognitive experiments on a laptop computer while sitting in bed. Every patient performed the experiment once, and
the total duration was approximately 90 minutes. Psychtoolbox was used to program the paradigm. Each experiment consisted of two
experimental sets, and each of these sets included two cognitive tasks in the following order: a valuation task and a two-alternative

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forced-choice task (2AFC). During the valuation task, high-resolution pictures of 20 junk food products (10 salty and 10 sweet) were
displayed on a laptop screen for 2 s in pseudo-random order. Patients were instructed to rate how much they wanted to eat the stimulus
on an arbitrary scale ranging from
100 to +100, in 8 steps of 25 units each. Responses were given by repeatedly pressing either the left
or the right key. The Likert scale was displayed until they pressed the OK button, without time restrictions. All answers were required to
be either a positive or a negative value. All stimuli were displayed three times in pseudo-random order, resulting in a total number of 60
trials per set. The 2AFC task consisted of a sequential presentation of two out of the same 20 stimuli. Each stimulus was displayed for 1
s. First, stimulus A was displayed for the first time (presentation A1), followed by stimulus B (presentation B), and finally stimulus A was
presented again (presentation A2). Patients were asked to report which stimulus they preferred and their confidence in each decision.
The Likert scale was displayed beneath both products, one on the left and one on the right, until the OK button was pressed, without
time restrictions. The scale was again divided in 8 steps of 25 units each: 4 steps for the right stimulus ranging from 25 to 100, and the
same for the left option. Responses were again given by repeatedly pressing either the left or the right key. The selection of the pair of
products and their temporal location as stimulus A or B was pseudo-randomly balanced. Stimulus location on the left or right during the
Likert scale was also pseudo-randomly balanced and independent of the temporal sequence of presentation. After the first set of these
two experiments, during a break, patients ate their preferred stimulus until they felt satiated. The preferred stimulus was defined as item
chosen most often in the 2AFC task, i.e., the item with the highest value ranking. In a second set of experiments, the participants
repeated the same tasks in the same order. During the 2AFC task, all possible combinations of products were presented once, resulting
in a total number of 190 different trials. For each trial, the sequence of presentation of the stimuli was maintained to ensure comparability
of trials between both experimental sets. The order of trials was pseudo-random and differed between experimental sets. Subjects were
informed during the break that after the second set of experiments they would have to consume again their preferred stimulus to sati-
ation. Note that one of the subjects declined to eat during the break, but their results were still included in our analyses since they did not
show any different behavior from the rest of the subjects (Figure S1B).

QUANTIFICATION AND STATISTICAL ANALYSIS

Mixed-effect hierarchical models

Linear mixed-effect models were used in the analyses of the behavioral data. Subject identity (12 levels) and the interaction between
subject identity and experimental session (24 levels) were introduced as random factors to reflect the nested structure of the data
(two experimental sessions within each subject). Every fixed-effect predictor was also introduced as a random slope on both random
factors, and a random intercept was always included. Furthermore, an unstructured variance-covariance matrix of the random ef-
fects was used to avoid restrictions in the estimation. Gaussian models with identity link function were estimated with the
maximum-likelihood method. Maximum-pseudolikelihood estimation was used for logistic regression models. All dependent and in-
dependent variables were standardized at the experimental set level. For inference, a Student’s t test of the fixed effect beta coef-
ficient versus zero was performed. Note that the models were not always the most parsimonious. Models were formally compared
using the Bayesian Information Criterion.

Responses were not biased by the design.

We tested whether our experimental design might have induced a bias in the direction of the decision, since we presented stimulus A
twice as often as stimulus B in any given trial. For each individual subject, we summed up the number of times that stimulus A was
chosen across all trials and the number of times that stimulus B was chosen across all trials. A signed-rank test of the number of
choices of A versus B including all experimental sets from all subjects showed no significant deviation of choice at the group level
(p = 0.4813, two-tailed signed-rank test). We performed a similar analysis for confidence and RT. For each subject, we calculated
the mean confidence across the trials in which stimulus A was chosen and for trials in which stimulus B was chosen. A signed-
rank test of confidence for choices of A versus B across all participants showed no significant difference at the group level (p =
0.3914). The same analysis was repeated for RT, for which we again found no significant difference for RT of A versus B (p =
0.3173). Altogether, we found no systematic bias induced by the presentation mode of stimuli A and B for choice, confidence, or re-
action time.

Similarity and change of cognitive variables

For any variable under consideration, the change between experimental sets for each variable was computed using Spearman’s cor-
relation between the variable during the first experimental task and the same variable during the second experimental task. In addi-
tion, the similarity between different cognitive variables was assessed for each pair of behavioral variables using the magnitude of the
Spearman’s correlation between both variables at the experimental set level. The magnitude of rho was appropriate since for variable
selection procedures of neuronal subpopulations the direction of the correlation is irrelevant.

Firing rate estimation and windows of analysis

For all analyses of the neural activity, the instantaneous firing rates were estimated using a trial-wise Gaussian convolution with a
standard deviation of 200 ms. Windows of analysis were defined to quantify the activity of each signal. In order to avoid an overlap
between the transient stimulus-locked signals and the persistent activity, the analysis of the persistent activity was restricted to the
epoch starting after the Likert scale presentation. The duration of the window was 2350 ms, which matched the shortest inter-trial

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duration throughout all experimental sets. This ensured that no overlap with the neural activity of the subsequent trial was included in
the window of analysis. For the transient neural activity, the window was defined between the onset and offset of each of the four
stimulus presentations (valuation, A1, B and A2), during both the valuation and the 2AFC tasks. All analyses were performed as a
sliding-window over the convoluted instantaneous firing rates.

Identification of relevant variables

A sliding-window Spearman’s rank-correlation was performed for each unit and each variable of interest, with a step size of 10 ms. A unit
was considered significant at a specific analysis step if it showed a significant rank-correlation between the trial-by-trial instantaneous
firing rate and the cognitive variable of interest during the first experimental set and if it simultaneously showed a significant correlation
during the second set at exactly the same step. Moreover, only those cells with the same sign of the correlation coefficient were accepted.
In order to obtainpa chance level of 5% of significantly correlated cells, the individual significance level of each of both correlations was
ffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi
corrected to a = 230:05, thus recovering a = 0.05 for each cell when the additional conditions of simultaneity and equidirectionality were
imposed. This univariate approach was selected due to the high number of variables and their multi-collinearity, which would yield un-
stable estimation of the effect of single predictors in a multiple regression, an approach similar to previous studies in decision making [52].
A correction for multiple comparisons was carried out to assess whether the proportion of significantly correlated neurons was
higher than expected by chance. A one-tailed binomial test was performed with an expected false positive probability of 0.05, match-
ing the alpha level of the correlation analysis. The proportion of significant units before the trial onset remained around chance level
(5%) for all analyzed variables, confirming that the assumptions of the binomial test were fulfilled. Finally, cluster-based permutation
tests were used to correct for multiple comparisons in the temporal domain [61]: 1000 simulations of each behavioral variable were
created with pseudo-random permutation and subsequently the correlation between the neural activity and each simulated data was
tested (a = 0.05, as mentioned above). The significance of the fraction of significantly correlated neurons at each time point was quan-
tified using the above-mentioned one-sided binomial test (a = 0.001) [null hypothesis: the fraction of correlated neurons was less than
or equal to the expected by chance (5%), alternative hypothesis: the fraction of correlated neurons was higher than expected by
chance]. Clusters were subsequently defined as consecutive time points reaching significance. For each cluster, the fraction of neu-
rons of each time point of the cluster were added, so as to include both the duration of the cluster and the magnitude of the effect in a
single measure. Based on this measure, the biggest cluster of each of the 1000 simulations was identified to create a distribution of
clusters. Finally, those clusters in the original data that were bigger than the tenth best cluster in this distribution were considered
significant (a = 0.01, one-sided permutation test) [null hypothesis: the size of the cluster was less than or equal to the clusters in
the simulated data, alternative hypothesis: the size of the cluster was greater than the clusters in the simulated data].
Visual features (red intensity, green intensity, blue intensity, hue, saturation, luminance and contrast) were extracted from the im-
ages with the Image Processing Toolbox. Hue and saturation were extracted with the rgb2hsv function from MATLAB. Luminance
was defined as the weighted sum of the color intensities (Luminance = 0.2126 3 red + 0.7152 3 green + 0.0722 3 blue) [57]. All visual
features, except contrast, were calculated as the average across all pixels in the image. Local contrast was defined as the standard
deviation of all the pixel intensities [62]. Nutritional features were defined as the content in grams of each of the stimuli (salt, sugar,
carbohydrates, calories, fat, fiber, and protein).

Estimates of subjective value were consistent

As an estimate of the valuation consistency within an experimental set, we computed the standard deviation (SD) of the valuation
ratings reported by the subjects during the three trials of each experimental set of the valuation task, and subsequently averaged
this SD across the 20 stimuli for each subject. The average valuation consistency across subjects was 14.67 ± 8.47 SD. Since
each discrete step in our Likert scale was of 25, this shows that subjects provided rather consistent valuation ratings on average.
Before the satiation break, the mean valuation consistency across subjects was 16.07 ± 9.68 SD, and after satiation 13.26 ± 7.22
SD. A signed-rank test of the mean valuation consistency before and after the satiation break yielded a p = 0.2661 (two-sided
signed-rank test). Thus, we assumed that these were fair estimates of value for the computation of decision variables. Moreover,
we also estimated the subjective values from the binary decisions in the two-alternative forced-choice task by ranking the times
that each item was chosen, thus reflecting the subjective values during the 2AFC task.

Variable update analysis

We tested whether the change in a behavioral variable from set to set was accompanied by correlated changes in neural activity. In
this analysis, the magnitude of the Spearman correlation coefficient between the actual variable and the firing rates (across trials) was
computed for each unit and averaged across both experimental sets (Actual). A surrogate version of the variable was used for which
the values of each tested variable were swapped between the experimental sets, meaning that firing rates from one set were now
correlated with the variable of the corresponding trials from the other set, i.e., the firing rates before satiation with the cognitive var-
iable after satiation and vice versa (Swapped).

Firing Rate 1st-Set Firing Rate 2nd-Set

Variable 1st-Set r11 r12
Variable 2nd-Set r21 r22

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where r11 and r22 are the actual correlation coefficients and r12 and r21 are the swapped terms. The magnitude of the correlation
was computed for each unit as
 
r11 ðjÞ + r22 ðjÞ

ActualðjÞ =   (Equation 1)
2 

where j is the neuron identity, and the magnitude of the surrogate as

 
r ðjÞ + r21 ðjÞ
SwappedðjÞ =  12 
 (Equation 2)
2
The idea behind this analysis is the following: for the units whose firing rate was correlated with a cognitive variable, e.g., subjective
value, if the variable change between experimental sets is reflected in the neural activity of the unit, the magnitude of the Actual cor-
relation (Equation 1) will be expected to be bigger than the magnitude of the Swapped correlation (Equation 2). Moreover, the covari-
ation of the variable change and the firing rate change should be reflected during the same time period during which the variable was
significantly correlated with the activity. Therefore, a sliding-window one-tailed Wilcoxon signed-rank test was performed to test
whether Actual versus Swapped [null hypothesis: Actual was less than or equal to Swapped, alternative hypothesis: Actual was
greater than Swapped). This analysis was first performed for all recorded units, thus avoiding circularity in the conclusion that could
arise from a selection of units to be analyzed. This analysis was then repeated with a subset of selected neurons. For the selection, all
neurons significantly correlated with the analyzed variable were included. Moreover, to avoid/reduce a selection bias, all cells signif-
icantly correlated with the swapped surrogate were also included. Both Actual-correlated and Swapped-correlated neurons selected
for the analysis were required to be simultaneously significant in both sets and equidirectional. Thus, this analysis tested for variable
update specifically for the variable-correlated units, excluding the non-related units from the analysis. Finally, cluster-based permu-
tation tests were once again used to correct for multiple comparisons in the temporal domain [61]: 1000 simulations of each behav-
ioral variable were created with pseudo-random permutations. The correlation between the neural activity and each simulated data-
set was then tested for both ‘‘Actual’’ and ‘‘Swapped’’ data. A Wilcoxon signed-rank test was performed as previously described (a =
0.05). Subsequently, clusters, i.e., consecutive time points reaching significance, were identified. For each cluster, the magnitude of
the Z-value of signed rank tests of each time point of the cluster were summed up, so as to include both the duration of the cluster and
the magnitude of the effect in a single measure. Based on this measure, the biggest cluster for each of the 1000 simulations was
identified to create a distribution of clusters. Finally, those clusters in the original data that were bigger than the tenth best cluster
in this distribution were considered significant (a = 0.01, one-sided permutation test).
As an alternative analysis, we analyzed for each neuron whether the change in firing rate between experimental sets correlated with
the change in a behavioral variable on a trial-by-trial basis. This analysis was again performed with a sliding-window Spearman’s
correlation (a = 0.05) and subsequently averaged across the time steps included in the window of analysis of the persistent activity
and the transient activity as defined above. The significance of the fraction of significantly correlated neurons was quantified using the
above-mentioned one-sided binomial test (a = 0.001) [null hypothesis: the fraction of correlated neurons was less than or equal to that
expected by chance (5%), alternative hypothesis: the fraction of correlated neurons was higher than expected by chance].

Quantitative comparison of variables

In order to find out which variables explained the highest degree of variance in the neural activity, quantitative model comparisons of
the persistent and the transient neuronal activity were performed. For this, variables were compared against the alternative models
using a post hoc binomial test. Only two variables were compared at each time step, e.g., confidence versus difference in value. The
binary comparisons were independent of other variables to ensure that adding more variables would not create complex patterns of
multi-collinearity during the variable comparison. If a third or more variables were simultaneously compared, these could share a
different amount of explained variance among them, thus biasing the selection toward those variables that shared less explained
variance, independently of their zero-order correlations. For units exhibiting a significant correlation with both variables under com-
parison simultaneously, each unit was assigned to the variable with the strongest effect size, defined again as the unsigned mean
correlation coefficient averaged across both experimental sets. The rationale behind this model selection was to sort the shared vari-
ability: if two variables showed an overlap in the identity of the correlated cells, we would expect that the better variable would have a
stronger correlation with more of these ambiguous neurons than the other would. This model selection was performed independently
for each time step, and subsequently the mean proportion of correlated neurons was averaged across all time steps in the window of
analysis and then rounded off to the next lower integer to yield an exact unit count for subsequent comparison. Finally, a one-tailed
binomial test was performed with an expected proportion of 0.5 (no preference) to determine whether the proportion of cells was
significantly higher for the variable of interest versus the control variable [null hypothesis: the fraction of correlated neurons with
the variable of interest was less than or equal to the fraction for the control variable, alternative hypothesis: the fraction of correlated
neurons with the variable of interest was greater than the fraction for the control variable].

Subpopulation and independence analyses

To elucidate whether the neural correlates of two cognitive variables were independent and/or correlated with segregated subpop-
ulations of neurons, three different analyses were performed.

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First, a model selection of each cell and each step of the sliding window was performed exactly as described in the previous sec-
tion: if a cell was significantly correlated with both cognitive variables simultaneously, the neuron was only counted for the variable
exhibiting stronger effect size, as described above. A one-tailed binomial test with an expected proportion of 0.5 (no preference) was
then performed to determine whether the proportion of cells was significantly higher for one variable versus the other.
In a second analysis, one of the variables was treated as a control variable. In a first step, all the units significantly correlated with
this control variable (e.g., confidence), were removed from the analysis. For remaining units, the proportion of units significantly corre-
lated with the other variable (e.g., RT), was calculated at each step. Importantly, units correlated with the control variable were not
removed from the denominator of the proportion. This allowed maintaining a common reference in the denominator, since the num-
ber of removed correlated cells changed over time steps. Moreover, if a control variable had a high proportion of correlated units,
removing these neurons not only from the enumerator, but also from the denominator could facilitate the statistical significance of
behavioral variables that were only weakly correlated with the control variable, similar to a suppressor effect in multivariable regres-
sion. Thus, with this procedure, we used a rather conservative criterion to establish statistical significance.
Third, we performed a sliding-window multivariate regression of the instantaneous firing rate for each cell and time point using the
two cognitive variables of interest (e.g., confidence and RT) as predictors for each neuron and time step. Both dependent and inde-
pendent variables were rank-transformed and Z-scored. The variable update analysis previously described was repeated using the
standardized regression coefficients.
The significance of the proportion of cells in the three analyses was tested using a binomial test during the windows of analysis, as
described above.

Changes of mind
Across all subjects, only in 12 trials out of a total of 1440 trials did a subject change their mind during the valuation task (0.83%). For
the 2AFC task, this was the case in 22 trials out of 4560 (0.48%). We therefore decided not to analyze them separately due to the lack
of sufficient data.

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