1. Reduced Repertoire of Cortical Microstates and Neuronal Ensembles in Medically Induced Loss of Consciousness 
Michael Wenzel, Shuting Han, Elliot H. Smith, Erik Hoel, Bradley Greger, Paul A. House, Rafael Yuste 
Cell Systems 
DOI: /j.cels
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2. Cell Systems DOI: (10.1016/j.cels.2019.03.007)
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3. Figure 1 Reduced Repertoire of Microstates in Mice during mLOC
(A) Awake, head-restrained mouse on a running wheel. Locomotion was measured by an infrared sensor. For seamless transitions across conditions, isoflurane was delivered through a custom tube placed right in front of the mouse. For LFP recordings, a pulled glass micro-electrode was carefully inserted into the cortex at around 250 μm depth through a small burr hole next to an implanted glass cover. Through the glass, two-photon calcium imaging was performed.
(B) Image of typical field of view and registered neuronal somata outlines (orange).
(C) Upper panel: representative brief raw LFP traces across five conditions. Lower panel: avg LFP delta range [1–4 Hz] spectral power across all five 10 min long conditions (n = 7 animals). Note the continuously increased delta power during surgical anesthesia.
(D) Superimposed locomotion of all 7 mice across conditions. Note that movement is absent in surgical and burst-suppression anesthesia.
(E) Calcium transients of 5 representative registered neurons across all five conditions.
(F) Corresponding raster plot of all registered neurons.
(G) Density map of microcircuit states, visualized by t-SNE for the entire experiment, displayed per condition. Vectors representing the population activity at each time point were transformed into a two-dimensional space while preserving local structures, and a density map was generated from the scatter plot.
(H) Quantification of locomotion across the entirety of each experiment (exp.), displayed as % locomotion per condition (10 min each); anesth., anesthesia.
(I) Neural activity level across conditions, quantified as probability of detecting activity within a moving 10 s window in a yes or no fashion; conditions colored as in Figure 1H.
(J) Relationship between LOC and neural activity during mild anesthesia versus recovery. Shown are the mean lags (n = 7 mice) of events (motion or spiking) with respect to the first (5 min) and second halves (5 min) of mild anesthesia or recovery. If motion or activity were evenly distributed throughout conditions mean lags would be 0. Note how locomotion consistently stopped in the first half of the mild anesthesia condition, while it re-emerged exclusively in the second half of the recovery condition (−90.6 ± 77.6 s versus 205 ± 48.6 s, Mann Whitney test [n = 7]; p = ). Importantly, in both conditions neural activity levels were much more evenly distributed across both conditions (−6.2 ± 7.8 s versus 14.2 ± 18.3 s, Mann Whitney test [n = 7]; p = 0.62).
(K) Boxplots of number of unique microstates (t-SNE/WS) across conditions as % of all identified unique microstates in a given experiment; line plots are individual experiments (grayscale).
(L) Total number of observed unique microstates (dashed lines) during wakefulness (red) or surgical anesth. (blue) versus corresponding distributions of values from 100 randomized datasets. (n = 7 mice, each exp. max-normalized for purpose of visualization). No overlap of observed versus random data (p < 0.01).
(M) Number of co-active neurons (in %, for purpose of visualization) versus the relative time spent by imaged population containing such co-activity, displayed per condition. Thick lines represent means, thin lines individual exp. conditions (N) Maximum number of co-active neurons (as % of all neurons per exp.) participating in a specific microstate, across conditions (colors as in M). Borderline statistical significance mild vs. surgical anesthesia p=0.062.
Figures 1H–1N show data from n = 7 mice; all error bars represent mean ± SEM; all boxes in boxplots represent 25–75 percentile of the data, bars within boxes represent means. Except comparison between observed and randomized data (Figure 1L) and comparison of two groups (Figure 1J), all statistical analyses represent one-way ANOVA with Bonferroni post-test. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p <
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4. Figure 2
Reduced Repertoire of Cortical Microstates and Ensembles during mLOC in Humans
All error bars in Figure 2 represent mean ± SEM (n = 2 patients).
(A) Photo of the craniotomy carried out on one patient in this study.
(B) Left: drawing of craniotomy in (A). Craniotomy (gray); arteries (red); veins (blue), multielectrode micro-array (MEA, black). Right: close-up photo of micro-electrode array (4 x 4 mm, 96 electrodes).
(C) Upper panel: representative brief raw LFP traces across three anesthetic conditions in one patient. Lower panel: avg LFP delta range [1–4 Hz] spectral power across conditions and patients.
(D) Raster plot of all single units in one patient.
(E) Two-dimensional density plot of same data after t-SNE, displayed per condition. Every dot represents a time point containing neural activity.
(F) Neural activity level across conditions, quantified as probability of detecting activity within a moving 10 s window in a yes or no fashion; conditions colored as in (C).
(G) Number of unique microstates (t-SNE/WS) across conditions as % of all identified microstates per exp.; line plots are individual patients.
(H) Number of unique microstates (APC) across conditions as % of all identified microstates per exp.; line plots are individual patients.
(I) Number of PCA components across conditions as % of all identified components per exp.; line plots are individual patients.
(J) LZW complexity across conditions; line plots are individual patients.
(K) LZW dictionary size across conditions; line plots are individual patients.
(L) Number of observed unique microstates (dashed lines) during mild anesth. (light blue) or surgical anesth. (dark blue) versus corresponding distributions of values from 100 randomized datasets, max-normalized for purpose of visualization. No overlap of observed versus random data, equal to p < 0.01; p1 and p2 represent patient 1 and 2.
(M) Number of co-active neurons versus the time spent by population in time points containing such co-activity per condition, displayed as mean across patients (thick lines). Thin lines represent conditions in individual patients.
(N) Maximum number of co-active neurons (as % of all neurons per exp.) participating in a specific microstate across conditions.
Cell Systems DOI: ( /j.cels )
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