Relationship between stimulus amplitude,stimulus frequency and neural damage during electrical stimulation of sciatic nerve of cat

The relation is investigated between stimulus frequency, stimulus pulse amplitude and the neural damage induced by continuous stimulation of the cat's sciatic nerve. The chronically implanted electrodes were pulsed continuously and the effects of the electrical stimulation were quantified as the amount of early axonal degeneration (EAD) present in the nerves seven days after the continuous stimulation. The primary effect of stimulating at 100 Hz rather than 50 Hz was to cause an increase in the slope of the plot of the amount of EAD versus stimulus lower. There was a small amount of EAD in three of the nerves stimulated at 20 Hz, but there was no detectable correlation between the amount of EAD and the stimulus amplitude. This suggests that continuous electrical stimulation of peripheral nerves at a low frequency induce little or no neural damage, even if the stimulus amplitude is very high. A preliminary presentation of the results has been made elsewhere (Agnew et al., 1993)     

Damage in peripheral nerve from continuous electrical stimulation: Comparison of two stimulus waveforms

The propensity for two types of charge-balanced stimulus waveforms to induce injury during eight hours of continuous electrical stimulation of the cat sciatic nerve was investigated. One waveform was a biphasic, controlled-current pulse pair, each phase 50 μs in duration, with no delay between the phases (‘short pulse’, selected to excite primarily large axons), whereas in the second type each phase was 100 μs in duration, with a 400 μs delay between the phases (selected to excite axons of a broader spectrum of diameters). The sciatic nerve was examined for early axonal degeneration (EAD) seven days after the session of continuous stimulation. With both waveforms, the threshold stimulus current for axonal injury was greater than the current required to excite all of the nerve's large axons. The correlation between simple stimulus parameters and the amount of EAD was poor, especially with the ‘short pulse’ waveform, probably due to variability between animals. When the stimulus was normalised with respect to the current required to fully recruit the large axons, a good association between damage and stimulus amplitude emerged. The damage threshold was higher for the ‘short pulse’ waveform. The implications for clinical protocols are discussed.     

An intrafascicular electrode for recording of action potentials in peripheral nerves

We are developing a new type of bipolar recording electrode intended for implantation within individual fascicles of mammalian peripheral nerves. In the experiments reported here we used electrodes fabricated from 25 μm diameter Pt wire, 50 μm 90% Pt-10% Ir wire and 7 μm carbon fibers. The electrodes were implanted in the sciatic nerves of rats and in the ulnar nerves of cats. The signal-to-noise ratio of recorded activity induced by nonnoxious mechanical stimulation of the skin and joints was studied as a function of the type of electrode material used, the amount of insulation removed from the recording zone, and the longitudinal separation of the recording zones of bipolar electrode pairs. Both acute and short term (two day) chronic experiments were performed. The results indicate that a bipolar electrode made from Teflon^™-insulated, 25 μm diameter, 90% Pt-10% Ir wire, having a 1–2 mm long recording zone, can be used for recording of peripheral nerve activity when implanted with one wire inside the fascicle and the other lead level with the first lead, but outside the fascicle. No insulating cuff needs to be placed around the nerve trunk.     

Estimation of the distribution of intramuscular current during electrical stimulation of the quadriceps muscle

Electrical stimulation is commonly used for strengthening muscle but little evidence exists as to the optimal electrode size, waveform, or frequency to apply. Three male and three female subjects (22–40 years old) were examined during electrical stimulation of the quadriceps muscle. Two self adhesive electrode sizes were examined, 2 cm × 2 cm and 2 cm × 4 cm. Electrical stimulation was applied with square and sine waveforms, currents of 5, 10 and 15 mA, and pulse widths of 100–500 μs above the quadriceps muscle. Frequencies of stimulation were 20, 30, and 50 Hz. Current on the skin above the quadriceps muscle was measured with surface electrodes at five positions and at three positions with needle electrodes in the same muscle. Altering pulse width in the range of 100–500 μs, the frequency over a range of 20–50 Hz, or current from 5 to 15 mA had no effect on current dispersion either in the skin or within muscle. In contrast, the distance separating the electrodes caused large changes in current dispersion on the skin or into muscle. The most significant finding in the present investigation was that, while on the surface of the skin current dispersion was not different between sine and square wave stimulation, significantly more current was transferred deep in the muscle with sine versus square wave stimulation. The use of sine wave stimulation with electrode separation distances of less then 15 cm is recommended for electrical stimulation with a sine wave to achieve deep muscle stimulation.     

Endoneurial extracellular matrix influences regeneration and maturation of motor nerve axons--a model of acellular nerve graft

Clinical results of functional reinnervation after application of autogeneous nerve grafts obtained from cutaneous nerves have not always been satisfactory. A foreign extracellular condition especially for regeneration of motor axons is assumed to be one of the reasons explaining these unsuccessful results. The role of endoneurial extracellular matrix in regeneration and maturation of motor axons was studied using acellular nerve segment prepared from muscular or cutaneous nerves applied between stumps of transected motor branch of the femoral nerve. No differences were found in the numbers of regenerated axons and related motoneurons through the motor and cutaneous nerve grafts 1 month after operation. Two months from grafting, however, the numbers of motoneurons and regenerated axons were increased significantly in the motor grafts while these were decreased after regeneration through the cutaneous grafts compared with 1 month. Axons' diameter and thickness of their myelin sheaths were similar in the cutaneous grafts 1 and 2 months after grafting. In comparison to 1 month, axons had larger diameter and thicker myelin in the motor than cutaneous nerve grafts 2 months from their application. Results of morphometric analysis indicate more beneficial extracellular conditions for regeneration and maturation of myelinated motor nerve axons in the acellular motor than cutaneous nerve graft. Generally, the results revealed that the endoneurial extracellular matrix of motor fibers has a positive effect on regeneration and maturation of motor nerve axons after lesion.     

Bioelectrical cochlear noise and its contralateral suppression: relation to background activity of the eighth nerve and effects of sedation and anesthesia

 The bioelectrical activity of the cochlea, without any ipsilateral acoustic stimulation, was recorded in awake guinea pigs (GPs) between electrodes chronically implanted at the round window (RW) and the skull. Measuring its power in the band centered around 1.0 kHz (0.5–2.5 kHz) provided an indirect measure of the ensemble background (EBA) activity of the eighth nerve. Contralateral white-noise (CLWN) stimulation reduced this EBA, presumably by activation of medial olivocochlear fibers. The aim of the investigation was to validate measurements of EBA and of its contralateral suppression in order to study the medial efferent function. The first goal was to find the best conditions for recording the EBA in the absence of ipsilateral stimulation and for studying its suppression by contralateral acoustic stimulation, which implies that no noise was generated by the experimental animal. Thus recordings were compared in normal, awake GPs and in GPs under sedation with xylazine, anesthetized with a combination of xylazine and ketamine, and with and without temperature regulation. In order to monitor the effects of sedation and anesthesia, the recordings were analyzed not only in the 0.5- to 2.5-kHz frequency band but also in the other frequency bands, 5–50 Hz, 50–150 Hz, and 150–500 Hz, which presumably include general central and neuromuscular contributions. The results show that sedation with xylazine accompanied by regulation of body temperature does not affect the EBA value nor its contralateral suppression. Nevertheless, anesthesia should be avoided, even with control of body temperature. The second goal of this study was to identify the specific cochlear contribution to the raw RW signal. Thus recordings were performed in normal and deafened animals and analyzed in the frequency band 0.5–2.5 kHz and also in the other frequency bands of 5–50 Hz, 50–150 Hz, and 150–500 Hz. The results indicate that most of the cochlear activity lies in the frequency band 0.5–2.5 kHz, with also some minor contribution coming from the 150- to 500-Hz band. Analysis and comparison of power values in the different conditions indicate that specific cochlear EBA power was about 60 μV². From a commonly accepted mean background discharge rate of 50 spikes/s (sp/s), the EBA power without CLWN should have been around 4.4 μV² if the fibers’ activity was random. This difference suggests that there is probably some degree of synchrony between individual fibers. There was a reduction of approximately 45% during CLWN stimulation. This suppression might correspond to a reduction in both discharge rate and synchrony of the fibers. Received: 28 January 1996 / Accepted: 10 April 1997     

Neuromuscular stimulation selectivity of multiple-contact nerve cuff electrode arrays

The feasibility of an electrical stimulation method selectively for activating skeletal muscles innervated by a common peripheral nerve trunk has been investigated. The method utilises ‘snugly’ fitting nerve cuffs that incorporate an array of 12 electrodes. These electrodes have been tested as four longitudinally aligned tripoles (located 90° apart on the cuff inner surface). In acute experiments on rabbit sciatic nerves, we have found that tripolar stimulation with this implant system is in general highly selective. ‘Field steering’, wherein a subthreshold transverse current is used in combination with a longitudinal tripolar current, tends to increase the selectivity of stimulation. On a rabbit sciatic nerve, a combination of adjacent longitudinal tripoles of the 12 electrode array generally yields a stimulation performance similar to that which would be expected if a 24 electrode array is used. This system may find a use in functional neuromuscular stimulation applications which require highly selective control over multiple muscles.     

Electrical activation of the human vestibulo-sympathetic reflex

Muscle sympathetic nerve activity (MSNA) is modulated on a beat-to-beat basis by the baroreflex. Vestibular input from the otolith organs also modulates MSNA, but characteristics of the vestibulo-sympathetic reflex (VSR) are largely unknown. The purpose of this study was to elicit the VSR with electrical stimulation to estimate its latency in generating MSNA. The vestibular nerves of seven subjects were stimulated across the mastoids with short trains of high frequency, constant current pulses. Pulse trains were delivered every fourth heartbeat at delays of 300–700 ms after the R wave of the electrocardiogram. Vestibular nerve stimulation given 500 ms after the R wave significantly increased baroreflex-driven MSNA, as well as the diastolic blood pressure threshold at which bursts of MSNA occurred. These changes were specific to beats in which vestibular stimulation was applied. Electrical stimulation across the shoulders provided a control condition. When trans-shoulder trials were subtracted from trials with vestibular nerve stimulation, eliminating the background baroreflex-driven sympathetic activity, there was a sharp increase in MSNA beginning 660 ms after the vestibular nerve stimulus and lasting for about 60 ms. The increase in the MSNA produced by vestibular nerve stimulation, and the associated increase in the diastolic blood pressure threshold at which the baroreflex-driven bursts occurred, provide evidence for the presence of a short-latency VSR in humans that is likely to be important for the maintenance of blood pressure during rapid changes in head and body position with respect to gravity.     

Induced hypothermia in peripheral nerve: Electron microscopic and electrophysiological observations

Summary Localized cooling of the cat sciatic nerve was achieved by application of a thermoelectric device to the exposed nerve at mid-thigh. The temperature of the nerve was maintained at 5°C for 2 h. Before, during, and after cooling, the response of the nerve to electrical stimulation was monitored, and compared with responses obtained in normothermic control preparations. In all cases, dropping the temperature of the nerve to 5°C resulted in total nerve block, and in all cases, function was restored when cooling stopped. Light and electron microscopic analysis of the cooled nerves revealed that despite functional recovery, a lesion affecting large myelinated axons developed over the course of 7 days. Both Waherian degeneration and segmental demyelination occurred. Unmyelinated fibres were not affected.     

Rapid axoplasmic transport of labelled proteins in the vagus and hypoglossal nerves of the rabbit

Summary Axoplasmic transport of labelled proteins was studied in the vagus and hypoglossal nerves of the adolescent rabbit after an intramedullary injection of ³H-leucine.As demonstrated by radioautography, the incorporated activity found in the intramedullary axons had a perikaryal origin and was transported distally at a rate of several mm per hour.The migration of labelled proteins along the axons outside the medulla was followed by measuring the specific radioactivity of the proteins in the hypoglossal and vagus nerves. A rapid transport of labelled proteins in proximo-distal direction was demonstrated at rates of 16–17 mm/h in the vagus nerve and 10–15 mm/h in the hypoglossal nerve. The effect of puromycin and nerve ligature on the rapidly moving component is described.     

Inhibition of midbrain-evoked tonic and rhythmic motor activity by cutaneous stimulation in decerebrate cats

The effect of mechanical and electrical stimulation of cervical cutaneous afferents was analysed on both the centrally induced tonic and rhythmic activities in hindlimb antagonist muscle nerves of 16 decerebrate paralysed cats. Electrical stimulation of dorsal midbrain evoked in the nerve to the tibialis anterior muscle (TAn) either rhythmic discharges (n=14), associated with tonic discharges in ten cats, or only tonic discharges (n=4). Centrally induced activity in the ipsilateral nerve to gastrocnemius medialis (GMn) occurred in fewer cats (n=12) and displayed similar patterns as in TAn. Manual traction of the scruff of the neck reduced the TAn tonic and rhythmic discharges (n=6) by 73% (P<0.05) and 71% (P<0.05), respectively, and reduced only the tonic component of GMn discharges (by 41%, n=3). Electrical stimulation (impulses 0.1–0.5 ms, 50 Hz) of cervical nerves belonging to C5 or C6 dermatomes, the intensity (0.4–4 mA) of which induced minimal inhibition of both TAn and GMn discharges, reduced significantly the tonic component of TAn discharges (by 39%, n=4). At higher intensities of electrical cervical nerve stimulation (2–6 mA) inducing maximal inhibitory effect, both tonic and rhythmic activities in TAn and GMn were both significantly reduced by, respectively, 81% and 94% in TAn (n=7), and by 49% and 43% in GMn (n=7). Electrical cervical nerve stimulation consistently reduced the isolated tonic discharge in TAn by 66% (n=4, P<0.05) and in GMn by 23% (n=3) when present. Thus the tonic component was more sensitive to inhibition than the rhythmic component of hindlimb muscle nerve activity. Electronic Publication     

Activity-dependent intracellular Ca2+ transients in unmyelinated nerve fibres of the isolated adult rat vagus nerve

 Confocal laser scanning microscopy was used to follow changes in the free intracellular calcium concentration ([Ca²⁺]_i) in nerve fibres and adjacent Schwann cells in isolated rat vagus nerves. [Ca²⁺]_i was monitored by the Ca²⁺-sensitive fluorescent dyes Calcium Green-1 and Fura Red. Intracellular Ca²⁺ transients were observed during repetitive (1–50 Hz) supramaximal electrical stimulation or by bath application of ATP. Trains of action potentials were more effective at elongated, fibre-like structures of the vagus nerves, whereas ATP-induced Ca²⁺ transients were found predominantly in regions of Schwann cell bodies. Activity-induced Ca²⁺ signals were unaffected by pharmacological manipulation of intracellular Ca²⁺ stores, during long-lasting application of purinergic receptor agonists, or by substitution of extracellular Na⁺ with Li⁺. However, they were abolished in the presence of Ca²⁺-free bathing solution or after the blocking of Ca²⁺ channels with Cd²⁺. Ca²⁺ transients were also observed during Ca²⁺ action potentials. Such ”Ca²⁺ spikes” were elicited by electrical stimulation in the presence of a combination of tetrodotoxin and K⁺ channel blockers. These data suggest that voltage-dependent Ca²⁺ channels, activated during short trains of Na⁺ action potentials, produce an increase in intra-axonal [Ca²⁺] of rat vagus nerves. We did not find evidence for activity-dependent Ca²⁺ transients in the Schwann cells surrounding the unmyelinated axons. Received: 16 September 1997 / Received after revision: 25 November 1997 / Accepted: 27 November 1997     

Schwann cells and endoneurial extracellular matrix molecules as potential cues for sorting of regenerated axons: A review

Besides very well elaborated microsurgical management of severed peripheral nerves, the clinical results of functional recovery following surgical repair of mixed nerves are disappointing. An improvement of functional recovery after peripheral nerve lesion requires the accurate regeneration of axons to their original target tissues and structures. Therefore, better clinical results could be obtained by a greater understanding of the cellular and molecular biology of selective nerve regeneration. The studies concerning Schwann cells and their endoneurial extracellular matrix as potent cues for selective promotion and influence of regenerating motor and sensory axons are reviewed. Knowledge of the sorting mechanisms of regenerated motor and sensory axons is needed not only for improvement of functional recovery, but also for the development of biocompatible nerve prostheses.     

Quantitative analysis of muscle hardness in tetanic contractions induced by electrical stimulation in rats

The purpose of this study was to investigate the in vivo relation between muscle hardness during an electrically induced contracting state and neuromuscular functions (M-wave and developed tension). Sixteen Sprague-Dawley rats were deeply anesthetized with urethane. Muscle hardness was measured quantitatively at the mid-portion of the gastrocnemius (GS) muscle during tetanic contractions induced by electrical stimulation (50 Hz, 100 μs duration) of the sciatic nerve or of the muscle directly. The M-wave was recorded with a pair of wire electrodes inserted into the muscle, and the developed tension was monitored with a push–pull gauge. Muscle hardness, M-wave amplitude and developed tension increased rapidly with the onset of nerve stimulation. Similar but intensity-dependent increases in muscle hardness and tension were observed following direct tetanic stimulation of the muscle. The hardness measured during nerve stimulation was correlated with the amplitude of the M-wave (r = 0.62, P < 0.0001) and the developed tension (r = 0.85, P < 0.0001). These phenomena were suppressed by pancuronium treatment (2 mg/ml, i.v.). These results suggest that muscle tension might be the most important factor for transcutaneously measured muscle hardness induced by tetanic muscle contraction.     

Facilitation of the withdrawal reflex by repeated transcutaneous electrical stimulation: an experimental study on central integration in humans

In the present human study, we aimed to investigate the facilitation of both the subjective pain responses, and the withdrawal reflex to consecutive transcutaneous electrical stimuli as measures of temporal summation. The frequency (0.5–20 Hz) and intensity (0.4–0.8 times the reflex threshold,  ×RT) of the electrical stimuli were systematically varied. When using repeated stimulation, the stimulus intensity that evoked pain was lower than that required by a single stimulus (temporal summation). Temporal summation leading to pain was found to depend significantly upon both frequency and intensity (e.g. stimulation at 1 Hz caused summation at 0.8 × RT, whereas stimulation at 20 Hz caused summation at 0.6 × RT). The strongest reflex facilitation, and hence the strongest pain intensity was obtained for stimulation at 10–20 Hz at an intensity of 0.8 × RT. In conclusion, the results of the present human study demonstrate clearly that a stimulus that is perceived as a localised, repetitive tactile tap can be integrated and cause severe pain. This suggests that pathologically generated sparse nociceptive afferent activity causes strong pain by central integration. This might be one mechanism to explain why clinical conditions can become excruciatingly painful despite the fact that the pathophysiological changes seem to be marginal (e.g. minor nerve trauma). Received: 26 April 1999 / Accepted: 12 August 1999     

Survival and regeneration of cutaneous and muscular afferent neurons after peripheral nerve injury in adult rats

Peripheral nerve injury induces the retrograde degeneration of dorsal root ganglion (DRG) cells, which affects predominantly the small-diameter cutaneous afferent neurons. This study compares the time-course of retrograde cell death in cutaneous and muscular DRG cells after peripheral nerve transection as well as neuronal survival and axonal regeneration after primary repair or nerve grafting. For comparison, spinal motoneurons were also included in the study. Sural and medial gastrocnemius DRG neurons were retrogradely labeled with the fluorescent tracers Fast Blue (FB) or Fluoro-Gold (FG) from the homonymous transected nerves. Survival of labeled sural and gastrocnemius DRG cells was assessed at 3 days and 1–24 weeks after axotomy. To evaluate axonal regeneration, the sciatic nerve was transected proximally at 1 week after FB-labeling of the sural and medial gastrocnemius nerves and immediately reconstructed using primary repair or autologous nerve grafting. Twelve weeks later, the fluorescent tracer Fluoro-Ruby (FR) was applied 10 mm distal to the sciatic lesion in order to double-label sural and gastrocnemius neurons that had regenerated across the repair site. Counts of labeled gastrocnemius DRG neurons did not reveal any significant retrograde cell death after nerve transection. In contrast, sural axotomy induced a delayed loss of sural DRG cells, which amounted to 22% at 4 weeks and 43–48% at 8–24 weeks postoperatively. Proximal transection of the sciatic nerve at 1 week after injury to the sural or gastrocnemius nerves neither further increased retrograde DRG degeneration, nor did it affect survival of sural or gastrocnemius motoneurons. Primary repair or peripheral nerve grafting supported regeneration of 53–60% of the spinal motoneurons and 47–49% of the muscular DRG neurons at 13 weeks postoperatively. In the cutaneous DRG neurons, primary repair or peripheral nerve grafting increased survival by 19–30% and promoted regeneration of 46–66% of the cells. The present results suggest that cutaneous DRG neurons are more sensitive to peripheral nerve injury than muscular DRG cells, but that their regenerative capacity does not differ from that of the latter cells. However, the retrograde loss of cutaneous DRG cells taking place despite immediate nerve repair would still limit the recovery of cutaneous sensory functions.     

双向脉冲可实现的神经纤维选择性兴奋

背景：研究证明利用电刺激外周神经纤维可恢复一些因失去中枢神经控制的肌肉的功能。 目的：验证双电极1 mm较近距离下双向方波脉冲实现神经选择性兴奋的正确性,并基于此实现神经的选择性兴奋。 方法：成年Wistar大鼠8只,麻醉后暴露大鼠坐骨神经,将电极小心放于坐骨神经干,建立神经选择性刺激模型。实验用电极为自制Cuff双极性电极,刺激器采用的是Grass S88刺激器和AWG2005任意波形信号发生器。采取双电极双向刺激方式,两个电极之间距离为1 mm,刺激波形选用脉宽为0.2 ms的对称双向脉冲,其输出脉冲的幅度、脉宽和延时均可调。调节刺激强度,研究双电极双向刺激下神经兴奋性的规律,以此实现神经的选择性兴奋,并利用“碰撞法”原理验证利用双电极双向刺激方法实现神经选择性兴奋的可行性。 结果与结论：实验过程中神经动作电位的变化将经P511放大器放大后接入示波器显示,双电极刺激波形为脉宽为0.2 ms的对称双向脉冲。随着刺激幅度的增大,实现神经的选择性兴奋。说明用距离很近(1 mm)的双电极双向对称脉冲的方法实现了神经的选择性兴奋,并利用“碰撞法”原理证实了此种方法的有效性和可行性。     

时间相干电场的外周神经无损刺激

外周神经电刺激可用于运动康复和慢性神经痛治疗,但目前具有空间选择性的无损刺激仍是一个有待解决的问题。提出一种基于时间相干(TI)电场的外周神经选择性无损电刺激方法,对大鼠坐骨神经进行实验,在其大腿腹侧与背侧皮肤上以平行于神经的方向布置刺激电极,通过相干电场扫描,将TI刺激峰值定位到神经上进行选择性刺激。结果表明,该方法可以在预先不知道神经确切位置的情况下通过扫描得出将刺激电场作用到神经的最佳电参数,从而实现对神经的选择性无损刺激,而且在刺激作用点不变的前提下实现刺激强度的控制。在此基础上研究TI电场对大鼠坐骨神经的刺激阈值I_T,测量固定频差Δf=0.5 Hz(n=12),改变频率f=1～6 kHz与固定f=5 kHz(n=11),改变频差Δf=0.5～10 Hz下的I_T,并将其和等幅kHz电场(n=7)的I_T进行比较。结果表明,等幅kHz电场的I_T显著高于TI电场(P<0.05),而且不同频率f下的I_T也有显著性差异(P<0.05),而不同频差Δf下的I_T却没有显著性差异(P>0.05),说明TI电流对大鼠坐骨神经的I_T受f影响而不受Δf影响,且刺激阈值I_T与频率f成正比关系。     

Regeneration of the rat spinal cord after thoracic segmentectomy: Growth and restoration of nerve conductors

The results presented in our previous report (Morfologiya, 127, No. 2 (2005)) provided evidence that consolidation of the spinal cord (SC) after thoracic segmentectomy in rats occurs as a result of the formation of a connective tissue scar, which is quicker when the defect is filled with collagen gel. The present report describes analysis of semithin sections and transmission electron microscopy studies demonstrating that regenerating nerve conductors traverse connective tissue in structures whose organization is identical to that of peripheral nerves. In the zone of SC rarefaction caudal to the trauma site, myelination of growing axons is mediated by glial cells without the formation of nerve trunks. Large numbers of fine regenerating conductors were seen at the sites of degenerated myelin fibers in the fasciculi of the white matter in the lumbar segments of the SC. __________ Translated from Morfologiya, Vol. 129, No. 1, pp. 30–38, January–February, 2006.     

Bands of Fontana are caused exclusively by the sinusoidal path of axons in peripheral nerves and predict axon path; evidence from rodent nerves and physical models

The precise cause of the bands of Fontana, striations on peripheral nerves visible to the naked eye, has been the subject of debate for hundreds of years. Some researchers have described them as reflecting the sinuous course of nerve fibres passing through nerves, and others have proposed that endoneurial collagen and sheaths surrounding nerves play a role in their appearance. We hypothesised that the bands are caused exclusively by reflection of light from the surfaces of nerve fibres travelling in phase in sinusoidal waveforms through peripheral nerves. We aligned images of obliquely illuminated nerves with confocal images of axons in those nerves, and the numbers and positions of the bands precisely matched the axonal waves. We also developed three‐dimensional models of nerves with representations of the sinusoidal path of axons at their surface. We observed patterns resembling the bands of Fontana when these models were obliquely illuminated. This provides evidence that the bands of Fontana can be caused by light reflected sinusoidal path of axons alone. We subsequently describe a mechanism of band production based on our observations of both nerves and models. We report that smaller diameter nerves such as phrenic nerves and distal branches of sciatic nerves have shorter band intervals than larger nerves, such as proximal trunks of sciatic nerves, and that shorter band intervals correlate with longer axons per unit length of nerve, which suggests a greater tolerance to stretch. Inspection of banding patterns on peripheral nerves may permit prediction of axon length within nerves, and assist in the interpretation of nerve conduction data, especially in diseases where axon path has become altered.