Ultrasound Guidance Combined with C-Arm Fluoroscopy in Selective Semilunar Ganglion Radiofrequency Thermocoagulation Through the Foramen Ovale for Trigeminal Neuralgia: A Randomized Controlled Trial.

OBJECTIVE: To explore the clinical value of ultrasound guidance combined with C-arm guidance during selective semilunar ganglion radiofrequency thermocoagulation via the foramen ovale for trigeminal neuralgia. METHODS: This study enrolled 48 patients diagnosed with trigeminal neuralgia between January 2021 and December 2021 in the Department of Pain Management at Xuanwu Hospital. Patients were randomly and equally divided into a C-arm-only group and an ultrasound-combined-with-C-arm (ultrasound+C-arm) group, according to a random number table. After exclusions, 42 patients were analyzed. Of these, 21 patients underwent selective semilunar ganglion radiofrequency thermocoagulation via the foramen ovale guided by the C-arm alone, whereas 21 patients underwent the same procedure guided by ultrasound combined with C-arm. The number of punctures, the amount of time elapsed until the target area of the semilunar ganglion was punctured, the cumulative dose of radiation exposure, and puncture-related complications were recorded during the operation. Numerical rating scale scores and radiofrequency thermocoagulation-related complications were evaluated preoperatively and at 1 day, 3 days, 7 days, 1 month, and 3 months after surgery. RESULTS: The number of punctures, the amount of time elapsed until the target area of the semilunar ganglion was punctured, and the cumulative dose of radiation exposure were all lower in the ultrasound+C-arm group than in the C-arm-only group (all P < 0.05). No significant differences were found in numerical rating scale scores and radiofrequency thermocoagulation-related complications between the two groups (P > 0.05). No puncture-related complications occurred in either of the groups. CONCLUSION: Ultrasound guidance combined with C-arm guidance could be safely used for puncturing the semilunar ganglion via the foramen ovale, with more efficiency and less radiation exposure than C-arm guidance alone.

Clinical study of puncture technique in single division of the trigeminal ganglion intumescentia.

OBJECTIVE: We explored the feasibility of single-division puncture in the ophthalmic division, maxillary division, and mandibular division of the trigeminal ganglion intumescentia (TGI) and the feasibility of radiofrequency treatment of trigeminal neuralgia. METHODS: According to the previous anatomical image studies, 3D Slicer software was used to analyze the CT images of the patients. The trigeminal ganglion fossa (TGF) was used as the imaging sign. TGI was identified in the sagittal plane along the fiber. The puncture path starts from the TGI center-foramen ovale line, extending outward to the epidermis as the needle insertion point, and extending inward to the division boundary. For lateral puncture, which is blocked by the mandible, the positions of closed mouth, open mouth, and over-open mouth were used. Multiple targets were generated using straight electrodes and curved electrodes to achieve full coverage of TGI. According to the preoperative design, general anesthesia surgery was performed. Xper CT was used for imaging, and the puncture was guided by Xper Guide. Radiofrequency treatment of TGI was conducted. RESULTS: In total, 45 patients with trigeminal neuralgia underwent 50 single-division TGI punctures. The procedure was smooth and the compliance with the design was good. Continuous radiofrequency (CRF) was performed, the VAS scores were 25 times at 70°C, 19 times at 65°C, two times at 60°C, and two times at 50°C (both in the ophthalmic division). Pulsed radiofrequency (PRF) was conducted two times. Within 24 h after the procedure, the VAS scores were all 0. From 1 to 7 days after the procedure, pain recurrence was found in three cases, of whom two cases received pulsed radiofrequency treatment. Patients were followed up for 1-24 months and there were no recurrence. After continuous radiofrequency at 65-70°C, the moderate tactile loss was observed, and nearly half of the patients had food residues on the surgical side after 6 months. After continuous radiofrequency at 60°C, there was mild tactile loss and no food residue. The tactile sensation was slightly decreased after continuous radiofrequency at 50°C, and the tactile sensation was normal the next day. CONCLUSION: Trigeminal ganglion intumescentia single-division radiofrequency is effective and feasible for the treatment of trigeminal neuralgia.

The impact of needle location on clinical outcome of radiofrequency rhizotomy for trigeminal neuralgia.

BACKGROUND: Radiofrequency thermocoagulation trigeminal rhizotomy (RT-TR) through the foramen ovale is a minimally invasive treatment for trigeminal neuralgia. Navigation of magnetic resonance imaging (MRI) and CT fusion imaging is a well-established method for cannulation of the Gasserian ganglion. In this study, we use the inline measurements from fusion image to analyze the anatomical parameters between the actual and simulation trajectories and compare the short- and intermediate-term outcomes according to determinable factors. METHODS: The study included thirty-six idiopathic neuralgia patients who had undergone RT-TR with MRI and CT fusion image as a primary modality or repeated procedures. RESULTS: Among thirty-six treated patients, the inline length of the trigeminal cistern was longer for the simulated trajectory (8.4 ± 2.4 versus 6.5 ± 2.8 mm; p < 0.05), and the predominant structure at risk extrapolated from the inline trajectory was the brainstem, which signified a more medially directed route, in contrast with the equal weighting of temporal lobe and brainstem for the actual trajectory. The preoperative visual analogue scale (VAS) was 9.3 ± 1.0, which decreased to 2.5 ± 2.6 and 2.9 ± 3.1 at first (mean, 3 months) and second (mean, 14 months) postoperative follow-up, respectively. The postoperative VAS scores at the two follow-ups were not statistically significant without a covariate analysis. After adjustment for covariate risk factors, the second follow-up sustained therapeutic benefit was evident in patients with no prior history of related treatment, an ablation temperature greater than 70 °C, and needle location within or adjacent to the trigeminal cistern. CONCLUSIONS: This preliminary study demonstrated that the needle location between cistern and ganglion also plays a significant role in better intermediate-term results.

Percutaneous Foramen Ovale Puncture: Usefulness of Intraoperative CT Control, in the Eventuality of a Narrow Foramen.

BACKGROUND: Balloon compression of the gasserian ganglion has been a well-established percutaneous treatment of trigeminal neuralgia since the 1980s. However, puncture of the foramen ovale by conventional single-plane fluoroscopy can be difficult in cases of local anatomic abnormalities. CASE PRESENTATION: We present the case of a 49-year-old woman diagnosed with idiopathic trigeminal neuralgia refractory to pharmacological treatment. After failure of puncture by conventional fluoroscopy for percutaneous gasserian ganglion balloon compression due to a narrow foramen ovale, the patient was submitted to puncture guided by computed tomography. CONCLUSION: Alternative imaging methods, such as computed tomography, should be considered when puncture of the foramen ovale by conventional single-plane fluoroscopy fails, to minimize the risk of potential complications triggered by frustrated puncture attempts.

Unconventional Facial Entry Points Confirmed Using a 3D CT Reconstruction-Guided Stereotactic Approach to Radiofrequency Thermocoagulation for the Treatment of Trigeminal Neuralgia: A Series of Case Reports.

OBJECTIVES: Patients with trigeminal neuralgia who are refractory to medical therapy may choose to undergo Gasserian ganglion percutaneous radiofrequency thermocoagulation. However, in cases where the foramen ovale is difficult to access due to various anatomical anomalies, the typical estimation of the facial entry point is suboptimal. METHODS: Three-dimensional computed tomography reconstruction imaging performed before surgery revealed anatomical variations in each of the four adult patient cases that made it more difficult to successfully access the foramen ovale (FO) for percutaneous radiofrequency thermocoagulation. Using measurements collected from preoperative imaging that showed each specific anatomical variation in the FO, researchers marked alternate facial entry points that would allow successful probe placement into the FO and recorded the arc angle data in the stereotactic instrument. RESULTS: Patients were evaluated during follow-up visits ranging from seven to 26 months after surgery and asked to rate postoperative pain using a visual analog scale. These scores decreased from 10 to 3 in all four patients by the third day after the procedure. There were no permanent complications or morbidities from the surgery. One patient experienced mild facial numbness; however, this side effect subsided within three months after surgery. During the follow-up period, no patient reported pain recurrence. CONCLUSIONS: The expectation for clinicians approaching trigeminal nerve block using a peri-oral approach should be to expect a great degree of potential variability in terms of both distances from the corner of the mouth and needle angle taken to successfully navigate the anatomy and access the foramen ovale.

Pro-nociceptive migraine mediator CGRP provides neuroprotection of sensory, cortical and cerebellar neurons via multi-kinase signaling.

Background Blocking the pro-nociceptive action of CGRP is one of the most promising approaches for migraine prophylaxis. The aim of this study was to explore a role for CGRP as a neuroprotective agent for central and peripheral neurons. Methods The viability of isolated rat trigeminal, cortical and cerebellar neurons was tested by fluorescence vital assay. Engagement of Nrf2 target genes was analyzed by qPCR. The neuroprotective efficacy of CGRP in vivo was tested in mice using a permanent cerebral ischemia model. Results CGRP prevented apoptosis induced by the amino acid homocysteine in all three distinct neuronal populations. Using a set of specific kinase inhibitors, we show the role of multi-kinase signaling pathways involving PKA and CaMKII in neuronal survival. Forskolin triggered a very similar signaling cascade, suggesting that cAMP is the main upstream trigger for multi-kinase neuroprotection. The specific CGRP antagonist BIBN4096 reduced cellular viability, lending further support to the proposed neuroprotective function of CGRP. Importantly, CGRP was neuroprotective against permanent ischemia in mice. Conclusion Our data show an unexpected 'positive' role for the endogenous pro-nociceptive migraine mediator CGRP, suggesting more careful examination of migraine prophylaxis strategy based on CGRP antagonism although it should be noted that homocysteine induced apoptosis in primary neuronal cell culture might not necessarily reproduce all the features of cell loss in the living organism.

A novel technique of delivering viral vectors to trigeminal ganglia in rats.

The objective of this study was to develop a viable and reliable technique of delivering viral vectors to rat trigeminal ganglia. Adult Sprague-Dawley rats (200-300 g) were used, and lentiviral vectors containing enhanced green fluorescence protein and calcitonin gene-related peptide short hairpin RNA (shRNA) were generated. Following general anesthesia, viral vectors were delivered to rat trigeminal ganglia using the technique described in this study. Both X-ray and micro-computed tomography (micro-CT) were employed to verify the position of the needles when injecting the vectors. In vivo fluorescence imaging and immunostaining against enhanced green fluorescence protein were performed to determine the success of viral transduction.The levels of calcitonin gene-related peptide in trigeminal ganglia were determined using real-time PCR, and pain levels following injections were evaluated using the Rat Grimace Scale. Our results show that injection needles can be advanced precisely at the trigeminal fossa and that viral vectors can successfully transduce trigeminal ganglia. Moreover, the levels of calcitonin gene-related peptide at trigeminal ganglia were down-regulated on day 7 after viral transduction. Pain levels returned to baseline by day 7 following injection. Therefore, we suggest that our trigeminal ganglion-targeting technique could be used for delivering genes or drugs to rat trigeminal ganglia.

Three-Dimensional Reconstruction of the Distribution of Neurons Contributing to Ophthalmic, Maxillary, and Mandibular Nerves in the Trigeminal Ganglion of Experimental Model.

The authors introduce a new method to build a three-dimensional (3D) model of the 3 branches of the trigeminal nerve in the trigeminal ganglion of rabbit with 3 different kinds of fluorescence. Ten adult New Zealand rabbits of both sexes weighing between 2.0 and 3.0 kg were used in the experiment. Then through an operation under general anesthesia, the maxillary and mandibular nerves were exposed, and red and gold fluorescence were applied to investigate the neurons of the maxillary and mandibular nerves. Subsequently, DiI was used as a marker for the ophthalmic neuron for the other side of the same rabbit. After receiving images of the 3 branches under a fluorescence microscope, a 3D model of the 3 branches of the trigeminal nerve could be built. The authors obtained an image of the 3 branches of neurons in the trigeminal ganglion, and a 3D model of the 3 branches of the trigeminal nerve in the trigeminal ganglion was reconstructed. In the trigeminal ganglion, ophthalmic neurons were concentrated in the anteromedial section, the maxillary division in the middle, and the mandibular division posterolaterally. Overlap was observed between the ophthalmic and maxillary neurons, and also for the maxillary and mandible neurons.

Gasserian Ganglion and Retrobulbar Nerve Block in the Treatment of Ophthalmic Postherpetic Neuralgia: A Case Report.

OBJECTIVES: Varicella zoster virus reactivation can cause permanent histological changes in the central and peripheral nervous system. Neural inflammatory changes or damage to the dorsal root ganglia sensory nerve fibers during reactivation can lead to postherpetic neuralgia (PHN). For PHN of the first division of the fifth cranial nerve (ophthalmic division of the trigeminal ganglion), there is evidence of inflammatory change in the ganglion and adjacent ocular neural structures. First division trigeminal nerve PHN can prove to be difficult and sometimes even impossible to manage despite the use of a wide range of conservative measures, including anticonvulsant and antidepressant medication. Steroids have been shown to play an important role by suppressing neural inflammatory processes. We therefore chose the trigeminal ganglion as an interventional target for an 88-year-old woman with severe ophthalmic division PHN after she failed to respond to conservative treatment. METHODS: Under fluoroscopic guidance, a trigeminal ganglion nerve block was performed with lidocaine combined with dexamethasone. A retrobulbar block with lidocaine and triamcinolone settled residual oculodynia. RESULTS: At 1-year follow-up, the patient remained pain free and did not require analgesic medication. CONCLUSION: To our knowledge, this is the first reported case of ophthalmic division PHN successfully treated with a combination of trigeminal ganglion and retrobulbar nerve block using a local anesthetic agent and steroid for central and peripheral neural inflammatory processes.

Clinical value of a self-designed training model for pinpointing and puncturing trigeminal ganglion.

OBJECTIVES. A training model was designed for learners and young physicians to polish their skills in clinical practices of pinpointing and puncturing trigeminal ganglion. METHODS. A head model, on both cheeks of which the deep soft tissue was replaced by stuffed organosilicone and sponge while the superficial soft tissue, skin and the trigeminal ganglion were made of organic silicon rubber for an appearance of real human being, was made from a dried skull specimen and epoxy resin. Two physicians who had experiences in puncturing foramen ovale and trigeminal ganglion were selected to test the model, mainly for its appearance, X-ray permeability, handling of the puncture, and closure of the puncture sites. Four inexperienced physicians were selected afterwards to be trained combining Hartel's anterior facial approach with the new method of real-time observation on foramen ovale studied by us. RESULTS. Both appearance and texture of the model were extremely close to those of a real human. The fact that the skin, superficial soft tissue, deep muscles of the cheeks, and the trigeminal ganglion made of organic silicon rubber all had great elasticity resulted in quick closure and sealing of the puncture sites. The head model made of epoxy resin had similar X-ray permeability to a human skull specimen under fluoroscopy. The soft tissue was made of radiolucent material so that the training can be conducted with X-ray guidance. After repeated training, all the four young physicians were able to smoothly and successfully accomplish the puncture. CONCLUSION. This self-made model can substitute for cadaver specimen in training learners and young physicians on foramen ovale and trigeminal ganglion puncture. It is very helpful for fast learning and mastering this interventional operation skill, and the puncture accuracy can be improved significantly with our new method of real-time observation on foramen ovale.

Development of a novel three-dimensional injection guide for trigeminal ganglia.

BACKGROUND: Trigeminal ganglion (TG) plays an important role in the process of orthodontic pain. It's necessary to design an accurate, precise and minimally invasive trigeminal ganglion injection guide plate to study TG. METHODS: Micro-CT was used to obtain the Dicom format data, and three-dimensional (3D) software (mimics and magics23.03) was used to reconstruct 3D head models. Design and modifications of the TG injection guide plate were performed in Magic 23.03 software, and the guide plate was produced by a 3D stereolithography printer. X-ray, micro-CT, Evans blue, and virus transduction were used to demonstrate the accuracy of the guide-assisted injection. Pain levels were evaluated after using the injection guide by a bite force test and Von Frey test. RESULTS: X-ray and micro-CT tests confirmed that the injection needle reached the bilateral trigeminal ganglia fossa. The Evans blue test and virus transduction proved that the injected drug could be accurately injected into the bilateral trigeminal ganglion and the lentivirus could be successfully transfected. The percentage of accurate injection was 10/10 (bilateral trigeminal ganglia). Orofacial pain induced by the trigeminal ganglion injection was mild and returned to baseline within seven days. CONCLUSION: The injection guide described in this study is viable and reliable for the delivery of drugs and virus transduction into the trigeminal ganglia.

Technique of Awake Computed Tomography-guided Percutaneous Balloon Compression of the Gasserian Ganglion for Trigeminal Neuralgia.

BACKGROUND: The classic percutaneous balloon compression (PBC) technique is used to complete an operation under the guidance of C-arm radiography under general anesthesia, making communication with patients during the operation impossible. It is not accurate or objective to predict the classic technique's curative effect solely by determining whether the projection of the x-ray lateral image of the filled balloon is pear-shaped. OBJECTIVES: This study aimed to upgrade classic PBC to awake computed tomography (CT)-guided PBC technology under conscious local anesthesia and analgesia monitoring. STUDY DESIGN: Prospective clinical study. SETTING: Department of Anesthesiology and Pain Medical Center, Jiaxing, People's Republic of China. METHODS: Puncture was designed and guided by CT scanning, and the curative effect was assessed by asking the patients about what they are feeling during the operation. RESULTS: CT can design the puncture path and accurately guide puncture, observe the position and shape of the balloon through 3-dimensional reconstruction during the operation, and judge the curative effect according to the patient's chief concern. LIMITATIONS: Local anesthetic analgesia is not perfect, resulting in some patients experiencing pain during surgery. CONCLUSIONS: PBC can be completed under conscious local anesthesia and analgesia. Its curative effect and operative end standard can be determined according to the patient's chief concern. Under CT guidance, the puncture path can be designed to complete an accurate puncture and to intuitively understand the position and shape of the balloon.

A feasibility study on pinpointing the branches of trigeminal nerve in radiofrequency ablation.

OBJECTIVE: To propose a pinpointing method and to obtain technique parameters for puncture of the branches of the trigeminal nerve through anatomical radiological study. DESIGN: Trigeminal ganglions and intracranial branches of 25 pieces (50 sides) of adult skull wet-specimens were dissected and coated with contrast agent. X-ray images of the skull in lateral cranial position were collected with the tube inclining towards head at 15°, 20° and 25°. 'Porus-Clinoid Line' and 'FO-PC axis' were delineated on the images. The latter set as the base line, Point A, B and C were settled separately as the upper rim of the trigeminal ganglion, the axis of Brach V2 and the junction of the extended border lines of Branch V2 and V3 all intersected with it. The collected data was processed afterwards. RESULTS: In the cases of 50 sides, the maximum value of the 'FO-PC Distance' was 17.8 mm; Distance A, 6.6 mm; Distance B, 10.1 mm; and Distance C, 6.6 mm, while the minimum of each was 9.4 mm, 0.3 mm, 4.4 mm and 6.6 mm respectively. Ratios of the 'FO-PC Distance' to Distance B were respectively 2.00 ± 0.15 mm, 1.98 ± 0.15 mm and 1.95 ± 0.16 mm when tube inclined towards head at angles of 15°, 20° and 25°; to Distance C were 3.06 ± 0.53 mm, 3.36 ± 0.60 mm and 3.75 ± 0.96 mm and to Distance A were 10.65 ± 9.17 mm, 7.33 ± 5.28 mm, 5.16 ± 2.30 mm under the same condition. CONCLUSION: The results showed that Distances from each branch of trigeminal nerve to the medial rim of foramen ovale vary on different individuals while the proportional relationship between each branch and 'FO-PC Distance' has regularity.

In vivo optical recordings of ion dynamics in mouse corneal primary nociceptive terminals.

This protocol aims to measure ion dynamics in nociceptive terminal endings in intact mice in vivo. We describe viral injection of GCaMP6s + RFP into trigeminal ganglia (TG) of mice, followed by calcium imaging of corneal nociceptive terminals that express GCaMP6s and RFP. This fast and high-resolution optical recording technique enables studying a nociceptive terminal's functional molecular network in physiological and pathological conditions. This platform can be applied to studying the physiology of terminals of other neurons. For complete details on the use and execution of this protocol, please refer to Goldstein et al. (2019).

Ultrasound localization microscopy and functional ultrasound imaging reveal atypical features of the trigeminal ganglion vasculature.

The functional imaging within the trigeminal ganglion (TG) is highly challenging due to its small size and deep localization. This study combined a methodological framework able to dive into the rat trigeminal nociceptive system by jointly providing 1) imaging of the TG blood vasculature at microscopic resolution, and 2) the measurement of hemodynamic responses evoked by orofacial stimulations in anesthetized rats. Despite the small number of sensory neurons within the TG, functional ultrasound imaging was able to image and quantify a strong and highly localized hemodynamic response in the ipsilateral TG, evoked not only by mechanical or chemical stimulations of corneal nociceptive fibers, but also by cutaneous mechanical stimulations of the ophthalmic and maxillary orofacial regions using a von Frey hair. The in vivo quantitative imaging of the TG's vasculature using ultrasound localization microscopy combined with in toto labelling reveals particular features of the vascularization of the area containing the sensory neurons, that are likely the origin of this strong vaso-trigeminal response. This innovative imaging approach opens the path for future studies on the mechanisms underlying changes in trigeminal local blood flow and evoked hemodynamic responses, key mechanisms for the understanding and treatment of debilitating trigeminal pain conditions.

Case report: computed tomography scan-guided Gasserian ganglion injection of dexamethasone and lidocaine for the treatment of recalcitrant pain associated with herpes simplex type 1 infection of the ophthalmic division of the trigeminal nerve.

We describe the case of a 17-year-old boy with dermatologic herpes simplex virus-1 outbreaks with incapacitating facial pain requiring multiple hospitalizations. He failed to respond to aggressive treatments including antiviral drugs, opioid analgesics, stellate ganglion, and supraorbital and supratrochlear nerve blocks. The patient elected to undergo dexamethasone and lidocaine Gasserian ganglion block under computed tomography-scan guidance. He had immediate and complete relief of his pain for the first time in almost 2 years. The patient remained pain free during 6-month follow-up visits. This is the first reported use of Gasserian ganglion block for treatment of herpes simplex virus-1 infection of the trigeminal nerve.

Rapid elevation of brain-derived neurotrophic factor production in the bilateral trigeminal ganglia by unilateral transection of the mental nerve in mice.

OBJECTIVES: Previous spinal nerve injury studies have reported brain-derived neurotrophic factor (BDNF) mRNA upregulation in either the ipsilateral dorsal root ganglion (DRG) neurons or both the contralateral and ipsilateral DRG neurons from early period after peripheral nerve injury. This BDNF elevation induces hyperalgesia in the injured and/or uninjured sites, but this detailed mechanism remains unknown. This study aimed to investigate the BDNF mRNA expression in bilateral DRG neurons caused by unilateral nerve injury and to explore the possible mechanisms by which nitric oxide (NO) mediates BDNF production in the DRG, resulting in contralateral hyperalgesia. METHODS: Early changes in BDNF mRNA expression in the bilateral trigeminal ganglia, within 1 day after mental nerve transection, were examined. Additionally, the effects on BDNF production of the NO synthase inhibitor N(ω)-nitro-l-arginine methyl ester (L-NAME) were investigated in the bilateral trigeminal ganglia. The relationship between injured neurons and BDNF production in the trigeminal ganglia was then assessed using immunohistochemical and retrograde tracing methods. RESULTS: Reverse transcription-PCR analysis demonstrated that unilateral transection of the mental nerve induced a rapid elevation of BDNF mRNA expression, which was inhibited by the intracerebroventricular administration of L-NAME prior to nerve transection. This effect was observed in both the ipsilateral and contralateral sides to the nerve transection. BDNF immunostaining combined with FluoroGold retrograde tracing revealed two types of BDNF-reactive neurons, FluoroGold-labelled and non-FluoroGold-labelled neurons, in the ipsilateral and contralateral sides of the trigeminal ganglia. BDNF-positive cells were also observed in the trigeminal ganglia of other trigeminal nerve branches. CONCLUSIONS: Unilateral nerve injury upregulates BDNF production in the bilateral trigeminal ganglia by NO-mediated and/or indirect activation of afferent neurons, resulting in contralateral hyperalgesia.

Stereotactic approach to the trigeminal ganglion using a stereotactic frame and intraoperative computed tomography scans: technical note.

OBJECTIVE: In this paper, a technique to place a probe at the trigeminal ganglion using a stereotactic frame and intraoperative computed tomography (CT) scans is described. METHOD: The procedure is performed on the CT table using a stereotactic frame. After the coordinates have been obtained and then adjusted, the target is probed using the frame. In this study, intraoperative scans were obtained to confirm the accuracy of probe placement. After successful stimulation studies, radiofrequency lesions were made. Thirty-two procedures were performed on 26 patients. RESULTS: Twenty patients were free of pain at a follow-up which lasted a median of 30 months. There was no permanent complication from the procedure. CONCLUSIONS: The technique is straightforward, accurate and safe. It also enables the surgeon to obtain intraoperative confirmation of accurate probe placement on CT images.

High-Throughput Calcium Imaging Screen of Toxins' Function in Dissociated Sensory Neurons.

Many toxins from a variety of venomous animals and plants have evolved to target neuronal ion channels and receptors. However, a significant obstacle in the study of these toxins is the finding and characterization of their specific molecular target. Here, we describe a method for fast and efficient screening of venom and toxin activity using live-cell calcium imaging. We describe the use of Fura-2, a calcium indictor that changes its fluorescence properties in response to intracellular calcium elevations, to measure the activity of neurons from the dorsal root and trigeminal ganglia. Calcium imaging is an efficient technique for testing many of the venom's components on large numbers of neurons simultaneously. This technique offers a novel tool for low-cost and rapid characterization of functionally active toxins and their target receptors.