The current action potential paradigm considers that all components beneath the neuron membrane are inconsequential. Filamentary communication is
less known to the ionic signal transmission; recently, we have proposed that the
two are intimately linked through time domains. We modified the atom
probe-connected dielectric resonance scanner to operate in two-time domains,
milliseconds and microseconds simultaneously for the first time. We resonate the
ions for imaging rather than neutralizing them as patch clamps do; resonant
transmission images the ion flow 10
The imaging of neural events has been one of the hallmarks of neurophysiology,
either by chemically neutralizing the ion channels, doping fluorescent molecules.
Thus far, the single ion channels [1,2] are mapped using 10
The electromagnetic (em) resonance of proteins in the milliseconds and nanosecond time domains has been measured since the 1930s [12-17] as kHz and GHz resonance frequencies, respectively. The report of GHz em resonance is much older than the millisecond ionic resonance of proteins first observed experimentally in the 1980s—through ionic spike was theoretically proposed in 1907 . In the current neuroscience, milliseconds signal transmissions are measured by neutralizing the ions in the patch-clamp. There is another way to do it, resonantly vibrating the ions and measuring transmitted signal (S21, S12 coefficients) across resonating ions. These two methods are very different. We chose the second protocol, which is not popular among neuroscientists. We use primarily Dielectric resonance microscopy, DRM, for the measurement introduced in 1995 [19,20]. The DRM is wireless, scale-free, looks deep inside a biomaterial; no chemical or physical contact is necessary. However, its potential to image the cell’s internal structures interacting in different time domains has been speculated but not explored yet. Thus far, just like in conventional biology, DRM has been used as a tool that maps a biomaterial in different time domains separately , as if different time domains are not connected.
To correlate different time domains into a singular architecture of time or clocks, identifying a typical biomaterial from the dielectric image is required. Here, we find em resonance as a critical biomarker so that ions, helices, secondary structures of proteins, DNA interacting at different time domains are mapped simply by changing the resonance frequency. Mapping the discrete clocks alone does not provide integrated information architecture of a biological system. A resonance frequency drives a system like a clock, several clocks arranged in a 3D shape build information architecture. For 40 years, time crystal research in biology tried to map biological events as nested clocks . We have advanced it further in neuroscience [23,24]. Time crystal is an ordered architecture of clocks model a self-operating biological system. As the clocks run, biological properties are generated.
Thus far, we have succeeded in observing self-similar fractal-like operations in three-time domains, milliseconds, microseconds, and nanoseconds. Those could be achieved by pumping kilohertz, megahertz, and gigahertz signals respectively to the biomaterial and looking into its reflected and transmitted signals surface profile . Recently, we showed that neural network circuits that we see under a microscope are not absolute . The isolated clusters of filaments located in distant neurons could wirelessly link, build circuits neglecting the synaptic pathways. Moreover, using quantum optics with electromagnetic resonance, we showed that at least three ordered structures inside a neuron build electromagnetic vortices, regulating ionic bursts of a neuron . One-to-one correspondence between neuron substructures and the vortex hologram generated by a neuron showed that transformation of electromagnetic to electric potential could happen. However, these observations are fairly abstract to conventional biology that is comfortable to see neuron communications in terms of spikes since 1907  and strongly founded on the finding by Hodgkin and Huxley that filaments inside a neuron are silent. Though contested, a map of sub-structure firing must be presented for a fair evaluation in competition with the membrane spike.
The filaments dispersed in the cell fluids were known to be silent, i.e., they
do not contribute to the potential of the membrane. However, when filaments are
packed in neural branches, e.g., axon initial segment, AIS, they might vibrate
like synchronized dipoles [23,24]. Filaments vibrating collectively could
generate more than threshold energy to affect membrane potential as a scale-free
resonance band connects proteins, filaments, and membranes . However,
detailed studies were required to find the geometric structure, whose corner
points are resonance frequencies. It is those multiple time domains connected in
a geometric shape, which hold invariants of neuron firing. We reported that
triplet of triplet resonance band, or a triangle whose corner points hold a
triangle inside. If the ionic transmission is blocked between a pair of neurons,
they still communicate . The possibility for two distinct communication
channels, electrical & ionic, is often explored in cell . Ionic and
non-ionic  transmissions together may lead to nonsynaptic
firing [29,30], endogenous firing [31,32]. For two decades,
electromagnetic resonance has been reported on the ion channels and filaments
regulating the firing [33-43]. However, simultaneously reading the
associated events unfolding at different time scales in a nerve spike is not done
yet. Our objective is to invent a tool to characterize both filament conductivity
& ion channel dynamics at a time in a cell, as they differ by 10
Though plenty of works on the mechanical resonance of neural or cellular fibers, few reports measure electromagnetic communications through the cell. Cell fluid damps the mechanical resonance since a mechanical vibration requires tension & physical motion. In contrast, fluid alone cannot dampen most parts of the electromagnetic spectrum since it requires rearranging the dipole, i.e., a pair of charges. Combining milliseconds’ membrane response with the dipolar and functional group responses in the nanoseconds-picoseconds time domains means connecting the ionic resonance with the dipolar resonance. However, connecting two diametrically opposite mechanisms is not easy. We need to image the activation and de-activation of ion channels with suitable tools, which did not exist even lately. Since an ion channel  opens/closes in 10–20 nanoseconds [44,45] we need a scanner that rapidly records (~10 ns) signals simultaneously at multiple time domains. The protein signals would only be recorded at a rate of their natural vibration as the nerve impulse transmits.
Moreover, proteins resonate at different time domains, transmitting ions only in milliseconds. How does this temporal management happen between host protein and guest ions? Our current work on quantum optics to create optical vortices of different angular momentums suggested many periodic structures made of proteins in the neuron. Monochromatic light shined on neurons creates a large condensate of vortices as a hologram. The hologram maps all the clocks; thus, the optical vortex hologram represents a 3D spatial arrangement of clocks or polyatomic time crystal. Many such clocks inside a protein probably regulate a protein operation in different time domains.
Rapid-frozen, cross-section image of an axon suggests an axon initial segment or AIS is filled with densely packed filaments [46-49]. The popular notion that the filaments float randomly in a fluid in the AIS is incorrect. Some filaments like microtubule are in parallel [46,47], unidirectional [50,51] with a gap ~50 nm , and nearly continuous  in the AIS. Outside AIS, the highly ordered Golgi apparatus isolates the filament-bundles at the axonal or dendritic branch junctions, filamentary continuity breaks. So, each part of the axonal or dendritic branches may appear continuous when seen from outside as a membrane. Inside, discrete, isolated filament bundles deliver a different character. However, the Golgi apparatus assists the filaments to remain parallel inside a branch similar to AIS . Each isolated filamentary bundle or a branch could act as a distinct electromagnetic resonator, similar to a tuning fork with multiple distinct resonance frequencies. Unfortunately, the ability of these isolated, independent resonators to absorb electromagnetic signals of particular frequency domains and emit like a separate antenna has not been explored. Our current work is the first attempt to map, theoretically model intricately, and experimentally verify how each branch between two junctions acts as a unique information processing device.
There is another intermediate structure between the filamentary core and the uppermost membrane layer. Just 2–3 nm below the membrane, actin, beta-spectrin form a periodic rectangular lattice-like structure . It is ~200 nm cross-sectional rings of beta-spectrin (BS) connected by parallel actin (A) wires (BSA). Together they build a crossbar architecture similar to the one we observe in integrated computer chips. The BSA-ordered structures are found almost everywhere of the neural network, in the glial cells and all dendritic, axonal branches of a neural network [55,56]. The BSA is not a singular structure, and it hosts various protein superstructures [57-59]. Several proteins are found to anchor with the grid junctions and specific locations. So, those proteins also become part of the global crossbar architecture. However, the ordering of guest protein molecules disappears if the microtubules in the central region of an axon are dissolved chemically . This is a very important development because it means Hodgkin and Huxley when melting the filaments in the 1950s, did destroy the cylindrical crossbar architecture wrapping the axonal and dendritic branches. The interfacing between the filamentary core and the membrane by highly ordered global crossbar architecture is not an accident. Multiple recent reports suggest that the network of filaments covers 98% by volume of an axon [46-49]. The filamentary core is bonded to the molecularly thin periodic actin-beta-spectrin hollow cylindrical network [54-60], directly in contact with the membrane above. That is why if the core melts, the crossbar architecture melts spontaneously together.
To this context, our recent work on probing the hippocampal neuron using polarized monochromatic light probed structural symmetry particulars of three distinct regions is important . Each structural symmetry provided a unique ring of light. However, our most important observation was imaging the energy transmission by filaments ignoring the membrane and other architectures across the neural network . The electromagnetic resonance field that emitted and absorbed energy from other non-connected neurons also fed the crossbar architecture. The crossbar architecture has 200 nm wide rings of proteins covering the cylindrical shape of the protein grid. A pair of such rings brighten up in the dielectric resonance image, which controls the ion channels’ opening and closing. Therefore, three layers create a triplet of triplet symmetry of clocks , and it’s a time crystal that we read using optical photon condensate .
Our earlier investigations did not isolate the contribution of each component; joint and superficial accounts were measured. Here, we have built a complete theoretical model of the three structures for the first time, matching theoretically predicted isolated and collective contributions using rigorous experiments. The most important finding reported here is that all prime contributors have threshold resonance frequencies that burst energy. So, we found that the dc potential burst of the membrane is the last or final event in a sequence of ac electromagnetic energy bursts. Membrane firing is not alone.
Based on the findings noted above, we build a model structure of AIS in the theoretical simulator computer science and technology, CST, particularly to justify that our scanner (Fig. 1a,b) could genuinely measure the signals from filaments & ion channels together. The core filament region is made of microtubules, and the neurofilaments have three parts. The first element of the dielectric structure is He, derived from the extended range of filaments throughout the axon . The second element is B, the unidirectional polarity of all filaments [50,51]. Third, Hi, an equidistant (~50 nm) lattice-like arrangement of parallel filaments [46-49] in the central core. Then we wrap the three types of dielectrics with the crossbar grid structure as the fourth component. The fourth component is Z, a periodic actin-beta-spectrin lattice coupled to the filamentary core [54-60]. Thus theoretically assembled dielectric material model, HeBHiZ representing an AIS. We cultured neurons and experimentally verified predicted energy exchange between distinct ordered structures of AIS. The tiny filaments vibrate at THz-GHz frequencies as a cascade of resonant oscillations builds the MHz periodic oscillations to assist the membrane’s kHz ionic spikes. HeBHiZ is the dielectric foundation of ionic bursts.
Basic concepts of SDIM used in the experiment. (a) Double piezo
scanner (Piezo1, Piezo2) based Scanning Dielectric Ion Microscope (SDIM) set up
for high-resolution non-contact imaging of a neuron membrane & its internal
structure. Diff. Amp., Differential amplifier; Inv Micro, Inverted Microscope.
(b) Basic scanning dielectric microscope SDM, which can perform as SDIM, an
easy-to-use nano surf STM, was converted to build 64 grid-based simultaneous
pixel data capture hardware. (c) He-ion microscopy of coaxial atom probe and its
schematic to the right, scale bar 250 nm (left). Schematic of a coaxial probe. (d)
An accurate contact is made to a rat hippocampal neuron using a coaxial probe,
scale bar 30
Now we describe the model construction in CST  in detail. The cytoskeleton has three kinds of filaments, microtubule, neurofilaments, and microfilaments (actin). We make all three components using pieces of dielectric materials of various elementary shapes. The elementary functional module’s geometric peculiarities are considered when choosing the elementary shapes to build a larger structure. For example, the spirally twisted cylindrical tube is used to emulate the shape of filaments and tube-based spiral loops to replicate the alfa-helices. Tape-like sheets are used to replicate beta-sheet protein-like structures. Then, by assembling these proteins, we build three kinds of filaments, actin, microtubule, and neurofilaments, of various diameters (5–10 nm). We also build ion channels of the membrane and lipid bilayer with similar dielectric properties.
Modeling of electromagnetic resonance of dielectric structures is a common
study. Our methodology of creating an artificial theoretical structure is novel
and was never attempted before. We do not create a replica of proteins by putting
blocks. Rather, we collect the protein structures from the PDB protein database
so that when we load them as dielectric helices and tapes in CST, we use an
accurate biomaterial structure. Here actin G, ankyrin, beta-spectrin, and tubulin
proteins build filaments. Using tubulin, we build a microtubule (25 nm
diameter). Using actin G, we build actin F filament or microfilament. Finally, to
build neurofilaments, five proteins, namely Peripherin, Internexin, Neurofilament
protein light (NPL), Neurofilament protein medium (NPM), and Neurofilament
protein-heavy (NPH), were used. The crystal structures of the neurofilaments do
not reveal an absolute composition of the five proteins. Therefore, we followed a
randomly chosen composition (1:1:3:3:3) to create 10 nm wide cylinders of various
stoichiometry. Neurofilaments have no definite composition & stoichiometry ,& they are not continuous like microtubules. Beta-spectrin size is kept
larger than the actin filament since the single-molecule resonance measurement
showed that it absorbs 10
In Fig. 2b, blue-colored beta-spectrin hetero dimers align end-to-end to form a complex tetramer that rolls around the central filamentary core like a ring. These beta-spectrin rings are cross-linked by short actin microfilaments extended along the length. This beta—spectrin—actin crossbar-grid binds directly to the plasma membrane; ankyrin protein resides in the grid but binds to the membrane proteins separately. So, the crossbar grid serves as a matrix of thin layer architecture holding many proteins in an ordered arrangement. The central core is made of continuous microtubules and discrete neurofilaments. Continuous microtubules mean that we find a single microtubule from the starting point to the end of AIS. Microtubules do not break in between, but neurofilaments do. The membrane is not part of our axon structure shown in Fig. 2b. We could remove different parts of AIS and simulate preferred frequencies for electromagnetic energy absorption, reflection, and transmission. Then we integrate filaments and proteins into the HeBHiZ structure, as shown in Fig. 2b, accurately following cryo-TEM derived parameters in computer simulation and technology (CST) simulator .
Measuring natural oscillations deep inside a neuron during
firing. (a) Schematic of a pair of coaxial probes entering synchronously first by
making contact with the membrane (7 nm wide) (top panel). Then, the coaxial probes
are inserted together using a piezo motor to contact the actin-beta-spectrin
periodic lattice (middle panel). The bottom panel shows the
microtubule-neurofilament core. A donut-shaped nerve spike (green) and a linear
plot (red) flow left to right as one surfs towards the bottom panel. (b) A
schematic of the simulated axon structure (8 rings, ~1.6
The final AIS structure (HeBHiZ) is shown in Fig. 2b. We added two energy supply ports at the two ends of the AIS structure for in silico measurement of reflection and transmission. Only one port is used for simulating the reflection coefficients. In this simulator, we solve Maxwell’s equation to derive the reflectance transmittance coefficients (S11 and S21) as a function of frequency. It is called the resonance spectrum.
We fused two types of already proven scanning microscopes used for over two decades into a new kind of microscope so that we could measure dipolar and ionic energy exchanges simultaneously. Two microscopes are the scanning ion conductance microscope (SICM, 1989) [9-11] and the scanning dielectric microscope (SDM, 1995) . We call it scanning ion dielectric microscope (SICM + SDM = SIDM, see Fig. 1a,b). One major change we made is to use the signal capture probe. We invented the world’s smallest patch-clamp to measure differential ac and dc conductance using three simultaneous channels at an extremely high signal-to-noise ratio . Since 2016, we have been reporting filamentary firing using these atomic-scale probes with an atomic-resolution coaxial electrode [64,65]. We must measure both neutralization of ions and dielectric resonance of ions because neutralization of ions has been a technique used since the early days of patch clamp-based action potential measurements. We have been arguing to replace the ion neutralization technique that measures neutralized ion content difference between the Soma’s cellular fluid and the buffer solution outside the neuron cell [63-65]. Instead, it is better to resonate the ions, keep them as the active component of the system, thus, do not interfere with the relative ionic balance of the cell and at the same time remotely measure reflectance or transmittance through the ionic clusters in a particular path using resonance spectroscopy. Another advantage of our method is that we could study wide ranges of materials simply by tuning and detuning probe ac signal frequency since each distinct biological material has a distinct resonance frequency peak distribution.
Therefore, in a SIDM, the signal source is a mixture of two frequencies: a millimeter-wave (GHz) to resonate with the measuring protein/molecule and a kHz wave to resonate with its released ions. When kHz resonance frequency syncs to read ions released from 0.4 nm hole of a 2 nm wide ion channel protein, the second frequency restricts the protein into a single conformer. Thus, the scan resolution increases from 100 nm [9,10] to 0.2 nm since the sharp atomic needle makes contact using a piezo motor (Fig. 1c,d). The atomic resolution has already been achieved using scanning dielectric microscopy , but not simultaneous recording in multiple time domains. In the SDIM setup (Fig. 1a,b), we use one coaxial probe where a Pt (0.1 nm) covered with a dielectric (e.g., glass) is wrapped by a cylindrical Au/Cr (100 nm) layer. The ratio of diameters between starting (d1) and endpoints (d2) of dielectric regulates the probe’s sensitivity. Two feedback loops run parallel to guide a probe’s piezo motor to make an atomic resolution contact with a single ion channel protein or a 2D surface scan. It records two signals simultaneously in two-time domains (500 ns and 50 ns) at a 10 ns time gap during approach or scan. So, we visualize ions that vibrate in the milliseconds time domain and dipoles that release ions in the nanoseconds time-domain together using a special setup (Fig. 1e; Fig. 2a).
Two significant technological developments were made, one in electronics and another in the tip. We also changed the ion density measurement protocol since we can truly measure a single ion channel or make contact with the membrane precisely.
Data capture rate: preamplifiers or lock-in amplifiers amplify a very low current ~1 pA but have an integration time of tens of microseconds. So, a firing event is recorded at a gap of 0.2 ms, misses any event happening in-between. If any event regulating a nerve spike begins and ends in microseconds, the existing neurophysiology characterization setups simply cannot detect such events. To achieve a real-time nanoseconds data acquisition, we use a 178 GHz (5.6 ps, ~0.2 THz) function generator connected to a monolithic chip MMIC operating at a THz speed (MMIC-THz). As a result, the integration of response pulses limits the data capture resolution to 10 ns (see below for details).
Tip dimension: The nanotubes (~20 nm diameter) are not
fit to read a single ion channel. 1
Current mode & voltage mode: When a glass tube clamps with a cell by removing patches from the membrane (Fig. 2e up), the measured current is due to the neutralization of ions. The measured voltage is a potential difference of cell fluid concerning the culture solution connected to the electrical ground. Here, we do not need to patch the probe with the membrane. We can truly measure the membrane. Moreover, our probe is so small that we can truly touch a single ion channel at the junction. Ions resonate with probe frequency, kHz (Fig. 2e). Therefore, we measure resonance current intensity as ionic current density passing through a single ion channel. Since there is no clamp, we use the term “current mode” and “voltage mode” instead of “current-clamp mode” and “voltage-clamp mode”, respectively. Coaxial probes driven by piezo motors are used in the SDIM in the voltage mode to read an ion channel on a ~7 nm thick membrane. In this mode, SDIM holds the constant current (Fig. 3b,c; Fig. 4a,b,c). The SDIM holds a fixed voltage (Fig. 2d; Fig. 4b—f). By varying the set frequency or editing the search frequency at the SDIM tip, one can contact the invisible & inaccessible components deep below a membrane.
Measuring natural oscillations deep inside a neuron during
firing. (a) Five electrodes are connected to an 8-day old rat hippocampal neuron
cultured from embryonic cells, S, Soma; D, Dendrimer; SA, Start of the axon; AIS, Axon
Initial Segment; EA, End of axon; AB, Axonal branch. Bottom, a microscope
image of a neuron, probes faded as the neuron is focused, scale bar is
Imaging & recording of single ion channel. (a) STM image of an
isolated membrane captured by rupturing a live neuron cell, at 2.3 V, 6 pA. The
scale bar is 7 nm. (b) SDIM image of a membrane captured by scanning a live neuron
cell at 3.3 V, 2 pA, 130 GHz. The scale bar is 16 nm. (c) For Na
A set of coaxial atom probes were connected to an active millimeter-wave monolithic integrated circuit (MMICs) based receiver and a transmitter module. Coaxial probes Au outer shell is fed with 178 GHz pulses, width 5.6 picoseconds, 120 pulsed responses are integrated for each value spending only 0.5 ns; we lose most time afterward. When 120 data was sampled at 220 GHz at the receiver, the integration for signal amplification by 3.4 dB reduced the resolution to 30 GHz, and noise filtering reduced the sampling resolution to 6 GHz. Thus, one ns time resolution in real-time data capture was obtained. A sub-harmonic 178 GHz local oscillator drove the Au tips of both probes, assembled with a broadband quadrature I/Q IF terminals, difference amplifier reduced I/Q imbalance below 1.8 dB, increasing output power above 3.5 dB. We read the change in potential at a gap of 10ns using a difference amplifier, and we get 100 more data between two consecutive 1ms readings.
Millisecond time-domain data were sampled at 40 kHz, amplified 300 times using a
low noise lock-in amplifier, and a multi-clamp 700B with digidata 1440A
(molecular devices A) data acquisition system, filtered using 10 Hz to the 50 kHz
bandpass filter. The peak-to-peak noise level of the device was (10
The coaxial probe used in this scanner reads an average signal from the
We followed the already reported neuron culture with minor changes [64,65].
The neuron culture plate was sterilized using 0.01% poly-l-lysine solution (5%
The neuron culture solution is prepared as follows: (1) First, we thaw the Single
Quots obtained from Lonza Inc at room temperature 300 K. (2) The basal medium is
added to the L-Glutamine and GA vials. (3) We add NSF-1 to the media to reach a
final concentration of 2%. Then aliquot the remaining NSF-1 to the desired
volume (e.g., 3
We removed a vial of embryonic neuron cells from the liquid Nitrogen chamber
We simulated the electromagnetic resonance of an artificial axon (Fig. 2b) by
varying its length. Dielectric resonance studies using Maxwell’s equations
delivered reflectance and transmittance. Anisotropic biomaterials split the
electromagnetic signal into asymmetric electric and magnetic field distributions
at resonance. Using this property, we provide evidence that if the past
experimental findings are true and follow them accurately, the axon structure
generates an MHz periodic oscillation naturally, by theory. At resonance
frequencies 3.8 THz, 5.5 THz, 7 THz, we simulate the spatial distribution of
electric and magnetic fields that periodically oscillates 360
All neuron data in this work are recorded on the 7–8 days old rat hippocampus neuron cells  grown by culturing the embryogenic cells on various substrates described earlier (Fig. 3a) [64,65]. The schematics below Fig. 3a show two types of contacts between the probe and the material. First, soft physical contact with the membrane surface (left) is shown. Second, the probe is inserted deep below the membrane (right). The potentials at different locations are plotted as a function of time (Fig. 3b,c). The locations are: (i) in the microtubule& neurofilament bundle in the central part of the axon tube (SA, EA), (ii) on the membrane of Soma (S), (iii) on the membrane of synaptic axon bouton (AB), (iv) actin-beta spectrin cylindrical net, located a few nm (3–5 nm) below the membrane (depicted as AcS) and (v) at the dendritic synaptic bouton (D). Fig. 3c shows four threshold potentials. First, AcS is the filament potential (FP). Second, SA and EA are the action initial segment’s potentials (AISP). Third, AB is the synaptic feedback potential (SP). These four potentials set the time of rising, sustain & fall of an action potential (AP). Here we depict AP as S. The SA & EA signals in Fig. 3b show two failed attempts to trigger a nerve impulse (depicted as search 1, 2) before their potentials reach a threshold to hold a flat potential. When SA and EA potentials are flat, energy flow reaches an equilibrium, and that causes the action potential to rise. It is an important observation. Since the technology does not permit us to observe events faster than a millisecond in the membrane-spike view of neuron firing, such signals remained unnoticed. The feedback potential AB acquires a more organized behavior under the wave train (Fig. 3c) than an isolated discrete pulse (Fig. 3b). Fig. 3c suggests that quantized energy transfer reveals multi-modal communications deep inside a neuron before a neuron fires. Pre-polarization is a fast spike with reverse polarity. It is followed by a nerve spike and is naturally observed in a patch-clamp (90 mV)  measurement. A reverse polarization is (~20 mV) not observed when a coaxial atom probe reads a nerve impulse. The observation is expected because the measurement mechanism of action potential here is not neutralization of cell fluids but measurement of dielectric property of ions or dipoles. Thus, observation is not limited by fluid flow.
Ion channels of the membrane are connected to a few proteins beneath, which link to the actin-beta spectrin cylindrical net that links the microtubule-neurofilament core. If we calculate the spatial ratio, 95% of a neural branch is densely packed with microtubule-neurofilament structure, i.e., HeBHiZ. Chemically, Hodgkin-Huxley has shown earlier in their Nobel-prize winning work  that neuron fires even if HeBHiZ does not exist. HeBHiZ is not essential for firing. However, our theory and experiment both argue that it edits the time gap between spikes, essential for brain cognition. If the microtubule-neurofilament structure is dissolved, then the membrane attached actin-spectrin periodic lattice-like architecture disappears. Their physical relationship was never measured before. Fig. 2d shows the normalized current (~10 pA) at 100 mV(RMS) ac signal (time = 1/frequency) for tubulin protein, microtubule-neurofilament core, actin-beta spectrin net, and the membrane —measured deep inside a neuron cell. The Gaussian distribution of around 80 measurements suggests that physically isolated neuron components are also temporally isolated. Since many measurements are averaged, we could conclude that five temporal regions exist. We reported the measurement of biomaterial’s triplet of triplet resonance bands in these materials earlier . Here, we measure the current response, not the reflected or transmitted pulse train, so it is an average current output of several proteins and complexes. By modifying the probe design, we could extend the operational time limit to record a wide range of elements in the axon. Since in the last 80 years, hundreds of publications reported dielectric resonance of many proteins, we could write a database of frequencies in the control software and detect proteins & structures if touched during a blind motion through the forest of proteins in an axon. We call it a protein hunt.
Ion channels do not need ATP (ambient temperature and pressure), so even an
isolated membrane slice open/close ion channels release ions. We scanned the
potential distribution for natural ion emissions from a membrane slice using a
Scanning Tunneling Microscope (STM) [23,24] (Fig. 4a). In SDIM imaging, the
lipid molecules of the membrane are not visible, which improves the ion channel
mapping (Fig. 4b). Thus far, a patch-clamp always recorded that a single ion
channel remains open or close for random durations. For a coaxial probe at tip
We measured the ion channel density for various lengths and diameters of the
axonal branches to confirm the unique ratio of the density of ion channels
Is it possible to regulate an ionic nerve impulse using the resonance frequencies of the components in the filamentary core of an axon that disrupts the three additional signals in Fig. 2b,c? Heating & cooling of a neuron is the EM effect , and heating changes the membrane potential . Infrared THz electromagnetic signals could edit the nerve impulse as an alternative tool to the ion channels [33-35]. A conduction failure could occur in an ion channel as a function of temperature. Neuron’s electromagnetic response is not limited to the infrared region only. A nerve emits MHz radio waves  and is extremely sensitive to microwave [41,42]. Fig. 5a shows that we apply a sub-threshold pulse to the Soma, not firing. We apply an electromagnetic signal of 22 MHz into the filament bundle of an axon core by bringing a Yagi antenna to its vicinity. The electromagnetic signal is applied in addition to the sub-threshold pulse train. Much to our surprise, the neuron exhibits a continuous above threshold ionic firing under a wave train of sub-threshold pulses.
Electromagnetic editing of a stream of the nerve impulse. (a)
20 mV stream of sub-threshold pulses applied to the neuron membrane (black). Two
output membrane readings are plotted with (red) and without (blue), applying an
additional ac frequency. Here, 12 MHz + 35 MHz + 7 GHz + 13 GHz is a set of ac
frequencies applied to the microtubule-neurofilament core using a coaxial probe
inside an axon core. (b) Above threshold 97mV pulse stream is applied along with a
set of ac frequencies (12 MHz + 35 MHz + 7 GHz + 13 GHz) to the
microtubule-neurofilament core (blue). The potential response on the membrane is
recorded by contacting the coaxial probe at Soma. (c) 5
Bidirectional electromagnetic control of the hypothalamus regulates feeding and
metabolism . Ca
Additional three signals that pass through the filaments are not essential for
the firing process, but they regulate the frequency modulation & the
multiplexing ability of a neuron via HeBHiZ. We reproduce the previous reports of
electromagnetic tuning of neural processes wirelessly with our new tool. The only
difference is that now we can say how the previous reports work in reality.
Finally, in Fig. 5c, the coaxial probe detects that a membrane’s internal
structure HeBHiZ has dipoles that oscillate and sends signals, even if there is
no firing. We placed two probes at two locations in the axon core and two on the
membrane (Fig. 2a), and by synchronizing the four probes as shown in Fig. 1e, we
determined that the natural pulses propagate as shown in Fig. 5c reaches the
In summary, we triggered a millimeter-wave to fire a neuron even using a
sub-threshold pulse where it should not fire by conventional wisdom 
(Fig. 5a). We could even stop the inevitable firing under an above threshold pulse
when mixed with a suitable millimeter-wave [84,85] (Fig. 5b). A membrane
fires even without the filaments inside. The filaments only modulate the spike
frequency . That is why even when the neuron does not fire, a natural
wave flows through the filaments (Fig. 5c). The whole neuron turns to an
integrated vibrating system where distant ion channels, irrespective of their
separation, are coupled to signal each other 10
Using a newly developed microscope SDIM, we could image even those ion channels
which are not transmitting ions. Our microscope read the dielectric resonance
signature by artificially sending signals and reading the reflected and
transmitted signals from the materials. Unlike light or infra-red waves, we use
the microwave and radio waves to bridge the milliseconds (10
The HH model  is part of the dynamics and interaction of electric charges of different temporal scales inside the neuron shaped by molecular interactions, regulating genes, protein expression. These phenomena remodel how electric interactions are performed and implicitly how information is processed and stored inside the neuron. They are required to achieve real-time information processing and bridge between the electrical nature of the brain (including AP generation) and intrinsic information processing at a molecular level. More importantly, the HH model does not describe the process of physical interaction when information is ‘read’,‘written’ or processed within the neuron. Our series of experiments indicate that signals and signs in the nervous system are information-rich . A single neuron is nested within and above the network of clocks, and geometry made by differential clocks holds the information as a polyatomic time crystal, suggesting that the brain is not a linear Turing tape but rather a fractal tape. The two concepts are orthogonal to each other.
AB designed the work; SG did the neuron work; PSi, SDK and KR did the axon theory; KS helped in the experimental set up of dielectric resonance; PSi, AB and PSa wrote the paper; JSM assisted in experiment and data analysis; RRP critically discussed the work.
Thank Dave Sonntag and Martin Timms for our device’s independent test & verification as part of patent US9019685B2.
Authors acknowledge the Asian office of Aerospace R&D (AOARD), a part of the United States Air Force (USAF), for Grant no. FA2386-16-1-0003 (2016–2019) on the electromagnetic resonance-based communication and intelligence of biomaterials.
The authors declare no conflict of interest.