Name: Establishment of a Modified Ferric Chloride-Induced Superior Sagittal Sinus Thrombosis (Video) | JoVE Journal
Uploaded: 2025-12-30T15:00:00.000+00:00
Duration: 7 min 34 s
Description: Learn a refined FeCl₃-soaked suture technique for inducing superior sagittal sinus thrombosis in rats with reduced cortical injury. This method enables reliable imaging and supports preclinical evaluation of cerebral venous thrombosis therapies.

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5 minutes

This work was supported by the National Natural Science Foundation of China (82401527).

This protocol has been approved by the Animal Experiments and Experimental Animal Welfare Committee of Capital Medical University with Approval No. AEEI-2025-138. The reagents and the equipment used are listed in the Table of Materials.
1. Establishment of a modified FeCl3-induced rat SSST model
<ol>
	<li>Use male Sprague-Dawley (SD) rats aged 6-7 weeks, weighing 220-250 g for the experiment. Prior to surgical procedures, acclimate the rats for 1 week and withhold food for 12 h.</li>
	<li>Prepare the following reagents, instruments, and equipment: 2% pentobarbital solution, 40% FeCl3 solution, 5% povidone-iodine, 75% alcohol, normal saline, ophthalmic ointment, disposable razor blades, scalpels, surgical scissors, forceps, 2-0 and 4-0 silk sutures, reverse-cutting needles, needle holders, 1-mL and 5-mL syringes, medical tapes, a rodent operating platform, a heating pad with rectal temperature probe, and a micro handheld cranial drill.</li>
	<li>After weighing, administer preoperative analgesia via subcutaneous injection of buprenorphine (0.1 mg/kg). Following a 15&#8211;20 min interval, anesthetize the rat by intraperitoneal injection of 2% pentobarbital (50 mg/kg) (following institutionally approved protocols). Then, pinch the skin of the hind limb with forceps to confirm the absence of withdrawal and pain responses.</li>
	<li>Apply ophthalmic ointment and shave the rat&#39;s scalp hair with a disposable razor blade.</li>
	<li>Secure the rat in a prone position on a rodent operating platform with medical tapes, and maintain the body temperature at 37 &#177; 0.2 &#176;C with a heating pad and continuously monitor core temperature with a rectal probe.</li>
	<li>Disinfect the scalp with alternating rounds of&#160;5% povidone&#8209;iodine, followed by 75% alcohol and repeat three times each.</li>
	<li>Make a 15-mm paramedian skin incision on the scalp, then bluntly dissect the underlying fascia and periosteum to fully expose the skull.</li>
	<li>Thin the skull with a cranial drill until the SSS is clearly exposed, starting from the lambda and extending 10 mm anteriorly along the sagittal suture. Employ an intermittent drilling technique to avoid damaging the underlying dura mater and SSS, and repeatedly irrigate the drill bit with normal saline to prevent thermal injury to the cortex.</li>
	<li>Apply a 10-mm segment of 2-0 silk suture soaked with 40% FeCl3 solution onto the exposed SSS surface for 5 min, then replace the suture with a newly soaked segment for another 5-min application.</li>
	<li>Rinse the surgical field with 0.5 mL normal saline three times to remove residual FeCl3.</li>
	<li>Close the skin with interrupted sutures using 4-0 silk suture, followed by povidone-iodine disinfection.</li>
	<li>Observe the rat until fully awake, then return the rat to the cage.</li>
</ol>
2. Assessment of successful model establishment
NOTE: During the surgical procedure, LSCI was used to measure the venous blood flow in the SSS after creating the cranial window and before FeCl3 application, and again after FeCl3 application and rinsing of the surgical field, but prior to skin closure. The model is considered successfully established when the venous blood flow in the SSS shows a significant decrease.
<ol>
	<li>Turn on the LSCI and the corresponding software (RFLSI v5.0), and then press online mode button.</li>
	<li>Adjust the height and position of the instrument until the indicator laser is centered in the field of view.</li>
	<li>Place the rat on a foam platform and adjust the position until the SSS aligns with the indicator laser.</li>
	<li>Adjust the magnification and focus until the image displays clearly. Set the lower and upper limits of the pseudocolor threshold to 10 and 200 to optimize the visual appearance of the pseudocolor image.</li>
	<li>Press the Set ROI button, select the circular tool, and delineate the SSS region.</li>
	<li>Press the Record button to obtain the venous blood flow and capture both original and pseudocolor images.</li>
</ol>
3. Assessment of thrombus area/volume and hemodynamic changes
NOTE: Several days to several weeks after the procedure, use high-resolution ultrasound to evaluate thrombus changes and vascular recanalization.
<ol>
	<li>Prepare the following drugs, instruments, and equipment: isoflurane, ultrasound gel, disposable razor blades, medical tapes, a small animal anesthesia machine, a high-resolution small animal ultrasound and photoacoustic imaging system, a 70 MHz ultra-high frequency UHF57x linear array transducer, and a 3D-acquisition motor.</li>
	<li>Install the 3D-acquisition motor and the UHF57x transducer. Then, initialize the motor.</li>
	<li>Anesthetize the rat in an induction chamber with 3% isoflurane mixed with 1 L/min oxygen.</li>
	<li>Place the rat in the prone position on the 37 &#176;C thermostatic imaging plate, with the head and limbs gently secured using medical tapes, and maintain anesthesia with 1.5%-2% isoflurane through a nose cone.</li>
	<li>Shave the scalp hair with a disposable razor blade to expose the detection area, then apply ultrasound gel.</li>
	<li>Adjust the position and orientation of the rat relative to the transducer until the SSS and intraluminal thrombus are clearly visualized.</li>
	<li>Acquire sequential tomographic images of the SSS in both sagittal and coronal planes using 3Dimaging in B-mode with a step size of 0.04 mm.</li>
	<li>Use Color Doppler mode to observe blood flow in the SSS, and then apply Pulsed-Wave (PW) Doppler mode to measure blood flow velocity.</li>
	<li>Discontinue anesthesia and return the rat to the cage after it is fully awake.</li>
	<li>Export the images and analyze them using Vevo LAB software. Record the maximum sagittal and coronal cross-sectional areas, and volume of the thrombus in the SSS, along with the maximum blood flow velocity of the sinus.</li>
</ol>

<ol><li>Saposnik, G., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Diagnosis+and+management+of+cerebral+venous+thrombosis%3A+a+scientific+statement+from+the+American+Heart+Association%5BTitle%5D)+AND+%22Stroke%22%5BJournal%5D)">Diagnosis and management of cerebral venous thrombosis: a scientific statement from the American Heart Association.</a> Stroke. 55 (3), e77-e90 (2024).</li>
<li>Cohen, O., Pegoraro, S., Ageno, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Cerebral+venous+thrombosis%5BTitle%5D)+AND+%22Minerva+Med%22%5BJournal%5D)">Cerebral venous thrombosis.</a> Minerva Med. 112 (6), 755-766 (2021).</li>
<li>Li, M., Chen, J., Meng, R., Ji, X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Rhythmic+deep-brain+magnetic+stimulation+promotes+angiogenesis+and+inhibits+apoptosis+by+regulating+the+JAK2%2FSTAT3+pathway+in+rats+with+superior+sagittal+sinus+thrombosis%5BTitle%5D)+AND+%22Brain+Res%22%5BJournal%5D)">Rhythmic deep-brain magnetic stimulation promotes angiogenesis and inhibits apoptosis by regulating the JAK2/STAT3 pathway in rats with superior sagittal sinus thrombosis.</a> Brain Res. 1852, 149509(2025).</li>
<li>Röttger, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+new+model+of+reversible+sinus+sagittalis+superior+thrombosis+in+the+rat%3A+magnetic+resonance+imaging+changes%5BTitle%5D)+AND+%22Neurosurgery%22%5BJournal%5D)">A new model of reversible sinus sagittalis superior thrombosis in the rat: magnetic resonance imaging changes.</a> Neurosurgery. 57 (3), 573-580 (2005).</li>
<li>Schoenwaelder, S. M., Jackson, S. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Ferric+chloride+thrombosis+model%3A+unraveling+the+vascular+effects+of+a+highly+corrosive+oxidant%5BTitle%5D)+AND+%22Blood%22%5BJournal%5D)">Ferric chloride thrombosis model: unraveling the vascular effects of a highly corrosive oxidant.</a> Blood. 126 (24), 2652-2653 (2015).</li>
<li>Hachenberger, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Aluminium+chloride+instead+of+ferric+chloride+for+inducing+superior+sagittal+sinus+thrombosis+to+reduce+ferromagnetic+artifacts+on+MRI+imaging+in+experimental+models%5BTitle%5D)+AND+%22Sci+Rep%22%5BJournal%5D)">Aluminium chloride instead of ferric chloride for inducing superior sagittal sinus thrombosis to reduce ferromagnetic artifacts on MRI imaging in experimental models.</a> Sci Rep. 14 (1), 10998(2024).</li>
<li>Gotoh, M., Ohmoto, T., Kuyama, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Experimental+study+of+venous+circulatory+disturbance+by+dural+sinus+occlusion%5BTitle%5D)+AND+%22Acta+Neurochir+%28Wien%29%22%5BJournal%5D)">Experimental study of venous circulatory disturbance by dural sinus occlusion.</a> Acta Neurochir (Wien). 124 (2-4), 120-126 (1993).</li>
<li>Mu, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+novel+rat+model+for+cerebral+venous+sinus+thrombosis%3A+verification+of+similarity+to+human+disease+via+clinical+analysis+and+experimental+validation%5BTitle%5D)+AND+%22J+Transl+Med%22%5BJournal%5D)">A novel rat model for cerebral venous sinus thrombosis: verification of similarity to human disease via clinical analysis and experimental validation.</a> J Transl Med. 20 (1), 174(2022).</li>
<li>Fries, G., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Occlusion+of+the+pig+superior+sagittal+sinus%2C+bridging+and+cortical+veins%3A+multistep+evolution+of+sinus-vein+thrombosis%5BTitle%5D)+AND+%22J+Neurosurg%22%5BJournal%5D)">Occlusion of the pig superior sagittal sinus, bridging and cortical veins: multistep evolution of sinus-vein thrombosis.</a> J Neurosurg. 77 (1), 127-133 (1992).</li>
<li>Wang, J., Ji, X., Ling, F., He, X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Comparison+of+anticoagulation+and+thrombolysis+treatments+in+a+rat+model+of+superior+sagittal+sinus+thrombosis%5BTitle%5D)+AND+%22Int+J+Neurosci%22%5BJournal%5D)">Comparison of anticoagulation and thrombolysis treatments in a rat model of superior sagittal sinus thrombosis.</a> Int J Neurosci. 124 (7), 532-541 (2014).</li>
<li>Ye, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Impacts+on+thrombus+and+chordae+Willisii+during+mechanical+thrombectomy+in+the+superior+sagittal+sinus%5BTitle%5D)+AND+%22Front+Neurol%22%5BJournal%5D)">Impacts on thrombus and chordae Willisii during mechanical thrombectomy in the superior sagittal sinus.</a> Front Neurol. 12, 639018(2021).</li>
<li>Chen, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Photothrombosis+combined+with+thrombin+injection+establishes+a+rat+model+of+cerebral+venous+sinus+thrombosis%5BTitle%5D)+AND+%22Neuroscience%22%5BJournal%5D)">Photothrombosis combined with thrombin injection establishes a rat model of cerebral venous sinus thrombosis.</a> Neuroscience. 306, 39-49 (2015).</li>
<li>Rahal, J. P., Malek, A. M., Heilman, C. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Toward+a+better+model+of+cerebral+venous+sinus+thrombosis%5BTitle%5D)+AND+%22World+Neurosurg%22%5BJournal%5D)">Toward a better model of cerebral venous sinus thrombosis.</a> World Neurosurg. 82 (1-2), 50-53 (2014).</li>
<li>Kim, D. E., Schellingerhout, D., Jaffer, F. A., Weissleder, R., Tung, C. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Near-infrared+fluorescent+imaging+of+cerebral+thrombi+and+blood-brain+barrier+disruption+in+a+mouse+model+of+cerebral+venous+sinus+thrombosis%5BTitle%5D)+AND+%22J+Cereb+Blood+Flow+Metab%22%5BJournal%5D)">Near-infrared fluorescent imaging of cerebral thrombi and blood-brain barrier disruption in a mouse model of cerebral venous sinus thrombosis.</a> J Cereb Blood Flow Metab. 25 (2), 226-233 (2005).</li>
<li>Srivastava, A. K., Kalita, J., Haris, M., Gupta, R. K., Misra, U. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Radiological+and+histological+changes+following+cerebral+venous+sinus+thrombosis+in+a+rat+model%5BTitle%5D)+AND+%22Neurosci+Res%22%5BJournal%5D)">Radiological and histological changes following cerebral venous sinus thrombosis in a rat model.</a> Neurosci Res. 65 (4), 343-346 (2009).</li>
<li>Wang, W., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Establishment+of+a+rat+model+of+superior+sagittal-sinus+occlusion+and+recanalization+via+a+thread-embolism+method%5BTitle%5D)+AND+%22Neuroscience%22%5BJournal%5D)">Establishment of a rat model of superior sagittal-sinus occlusion and recanalization via a thread-embolism method.</a> Neuroscience. 416, 41-49 (2019).</li>
<li>Hachenberger, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Comparison+of+edoxaban+and+enoxaparin+in+a+rat+model+of+AlCl3-induced+thrombosis+of+the+superior+sagittal+sinus%5BTitle%5D)+AND+%22Naunyn+Schmiedebergs+Arch+Pharmacol%22%5BJournal%5D)">Comparison of edoxaban and enoxaparin in a rat model of AlCl3-induced thrombosis of the superior sagittal sinus.</a> Naunyn Schmiedebergs Arch Pharmacol. 396 (12), 3887-3892 (2023).</li>
</ol>

The authors have no conflicts of interest to declare.

CVT is an uncommon but potentially life-threatening specific type of cerebrovascular disease1. To further investigate its pathological mechanisms and develop effective intervention strategies, researchers have established various animal models of SSST, primarily including SSS ligation or occlusion7,8,9, prothrombotic substance injection10, autologous blood clot injection<sup class=...

Cerebral venous thrombosis (CVT) is a special type of cerebrovascular disease, accounting for 0.5-3% of all strokes and serving as an important cause of stroke in young adults1. The core pathological change of this disease is thrombosis within the dural venous sinuses and/or cerebral veins, which leads to impaired cerebral venous drainage, subsequently causing increased intracranial pressure, cerebral edema, venous infarction, or hemorrhage2. To better elucidate its underlying mechanisms and evaluate novel therapeutic interventions, reliable and reproducible animal models are indispensable. Given that the superior sagittal sinus (SSS) is the most frequently affected site and its superficial location facilitates operational procedures2,3, current animal experiments primarily utilize the superior sagittal sinus thrombosis (SSST) model.
Among various modeling methods, the ferric chloride (FeCl3) induction method has become one of the most widely used techniques due to its operational simplicity and cost-effectiveness4. In this model, the topical application of a FeCl3 solution directly damages the venous sinus endothelium, exposing the subendothelial matrix through its strong oxidative properties. This exposure activates platelet aggregation and the coagulation cascade, ultimately inducing venous sinus thrombosis5. However, compared with other methods, the traditional FeCl3-filter paper model also has several drawbacks. The most critical issue is that the paramagnetic properties and high magnetic susceptibility of FeCl3 generate substantial artifacts during magnetic resonance imaging, severely compromising the accuracy of thrombus assessment and venous sinus recanalization observation6. Furthermore, the shape of the filter paper does not fit well with the SSS, making it prone to injuring the brain parenchyma surrounding the sinus. Therefore, it is difficult to determine whether the parenchymal damage around the SSS results from the thrombosis itself or from direct chemical injury caused by FeCl3.
The overall goal of the present work is to establish a modified FeCl3-induced SSST rat model that reduces direct cortical injury caused by FeCl3 and allows for the assessment of blood flow and thrombus size within the SSS. This study introduces a paramedian approach combined with a FeCl3-soaked suture technique, which minimizes direct cortical chemical injury caused by FeCl3, and enables real-time, noninvasive, and quantitative monitoring of venous sinus blood flow and thrombus formation using high-resolution ultrasound.

<table><tbody><tr><td>1 mL Syringe</td><td>Shenzhen Huayon Biotechnology Co., Ltd.</td><td>21-3020</td><td></td></tr><tr><td>2-0 silk sutures</td><td>Shanghai Pudong Jinhuan Medical Products Co.,Ltd</td><td>A224</td><td></td></tr><tr><td>3D-acquisition motor</td><td>Fujifilm VisualSonics</td><td>https://www.visualsonics.com/gallery/extended-3d-motor</td><td></td></tr><tr><td>4-0 silk sutures</td><td>Shanghai Pudong Jinhuan Medical Products Co.,Ltd</td><td>A222</td><td></td></tr><tr><td>5% povidone-iodine</td><td>Shandong Lircon Medical Technology Co., Ltd.</td><td>https://www.lircon.cn/products_details/100.html</td><td></td></tr><tr><td>5 mL Syringe</td><td>Shenzhen Huayon Biotechnology Co., Ltd.</td><td>21-3023</td><td></td></tr><tr><td>Disposable razor blade</td><td>Dongyang Jinmin Trading Co., Ltd.</td><td>ST300</td><td></td></tr><tr><td>FeCl&lt;sub&gt;3&lt;/sub&gt;</td><td>Macklin</td><td>I811935</td><td></td></tr><tr><td>Heating pad</td><td>Shenzhen Huayon Biotechnology Co., Ltd.</td><td>36-0001</td><td></td></tr><tr><td>Isoflurane</td><td>RWD Life Science</td><td>R510-22-10</td><td></td></tr><tr><td>Laser speckle imaging system</td><td>RWD Life Science</td><td>RFLSI III</td><td></td></tr><tr><td>Linear array transducer</td><td>Fujifilm VisualSonics</td><td>UHF57x</td><td></td></tr><tr><td>Medical tape</td><td>Suzhou Konlida Medical Supplies Co., Ltd.</td><td>B02</td><td></td></tr><tr><td>Micro handheld cranial drill</td><td>RWD Life Science</td><td>78001</td><td></td></tr><tr><td>Needle holder</td><td>RWD Life Science</td><td>F31025-13</td><td></td></tr><tr><td>Normal saline</td><td>Shanghai Beyotime Biotechnology Co., Ltd.</td><td>ST341-500ml</td><td></td></tr><tr><td>Ophthalmic ointment</td><td>Shenzhen Huayon Biotechnology Co., Ltd.</td><td>19-7223</td><td></td></tr><tr><td>Reverse-cutting needles</td><td>Shanghai Pudong Jinhuan Medical Products Co.,Ltd</td><td>TJ-3-514</td><td></td></tr><tr><td>Rodent operating platform</td><td>Shanghai Yuyan Scientific Instrument Co., Ltd.</td><td>30150</td><td></td></tr><tr><td>Scalpel blade</td><td>RWD Life Science</td><td>S31015-01</td><td></td></tr><tr><td>Scalpel handle</td><td>RWD Life Science</td><td>S32003-12</td><td></td></tr><tr><td>Small animal anesthesia machine</td><td>RWD Life Science</td><td>R500</td><td></td></tr><tr><td>Small animal ultrasound and photoacoustic imaging system</td><td>Fujifilm VisualSonics</td><td>Vevo F2</td><td></td></tr><tr><td>Smooth Forceps</td><td>RWD Life Science</td><td>F12013-10</td><td></td></tr><tr><td>Sodium Pentobarbital</td><td>Sigma-Aldrich (Shanghai) Trading Co., Ltd.</td><td>P3761</td><td></td></tr><tr><td>Surgical Scissors</td><td>RWD Life Science</td><td>S14014-10</td><td></td></tr><tr><td>Toothed Forceps</td><td>RWD Life Science</td><td>F12011-13</td><td></td></tr><tr><td>Ultrasound gel</td><td>Shandong Lircon Medical Technology Co., Ltd.</td><td>https://www.lircon.cn/products_details/230.html</td><td></td></tr></tbody></table>

establishment of a modified ferric chloride-induced superior sagittal sinus thrombosis

Establishment of a Modified Ferric Chloride-Induced Superior Sagittal Sinus Thrombosis

Cerebral venous thrombosis (CVT) is a rare but potentially life-threatening neurological disorder. Animal models offer critical platforms for investigating their pathophysiology and evaluating potential therapies. In this study, we present a modified rat model of superior sagittal sinus (SSS) thrombosis using a FeCl&#8323;-soaked suture technique. Male Sprague-Dawley rats (220-250 g) underwent a paramedian scalp incision and creation of a 1 cm cranial bone window over the SSS. Compared to the traditional midline incision, this approach prevents adhesion among the incised skin, the bone window, and the underlying dura mater. Additionally, it avoids the interference caused by suture knots located over the SSS, thereby improving the quality of ultrasound imaging. A 2-0 silk suture soaked in 40% FeCl3 was applied directly onto the exposed sinus for two consecutive 5-min intervals. Compared to the traditional FeCl3-filter paper method, this technique minimizes direct chemical injury to the parasagittal cortex. Laser speckle contrast imaging was employed to assess cerebral venous perfusion before and after FeCl3 application. A significant reduction in blood flow confirmed successful thrombosis. Seven days post-surgery, thrombus formation and hemodynamic changes were evaluated via high-resolution ultrasound and analyzed using Vevo LAB software, which provided volumetric and hemodynamic data of the thrombus. This modified model offers a reproducible, minimally invasive, and imaging-compatible approach for preclinical CVT research.

The schematic diagram of the modified FeCl3-induced SSST model is shown in Figure 1. Experimental results demonstrated that after application of the FeCl3-saturated silk suture to the SSS, LSCI detected a significant reduction in local blood flow, indicating successful occlusion of the venous sinus and confirming the sensitivity of LSCI for detecting dynamic perfusion changes (Figure 2). On postoperative day 7, small-animal ultrasound confi...

This study presents a protocol for establishing a reproducible, minimally invasive rat model of superior sagittal sinus thrombosis induced by ferric chloride, utilizing a silk-suture method in combination with laser speckle contrast imaging and high-resolution ultrasound for evaluation.

Cerebral venous thrombosis (CVT) is a rare but potentially life-threatening neurological disorder. Animal models offer critical platforms for ...

In our field, several modeling methods for superior sagittal sinus thrombosis are used. However, the ferric chloride induction method is widely used due to its simplicity and the low cost. We aim to refine the ferric chloride-induced superior sagittal sinus thrombosis model by reducing cortical injury and visualizing sinus thrombus.To begin, apply ophthalmic ointment to both eyes of an anesthetized rat and shave the scalp hair carefully. Then, secure the rat in a prone position on a rodent operating platform using medical tape. Maintain the body temperature at around 37 degrees Celsius with a heating pad.Disinfect the shaved scalp thoroughly using 5%povidone-iodine, followed by 75%alcohol, and repeat three times each. After confirming anesthetic depth by toe pinch reflex, make a 15-millimeter peri-median skin incision on the scalp. Using blunt dissection, separate the underlying fascia and periosteum to expose the skull fully.Thin the skull using a cranial drill until the superior sagittal sinus is clearly exposed, starting from the lambda and extending 10 millimeters anteriorly along the sagittal suture. Employ an intermittent drilling technique to avoid damage to the underlying dura mater and superior sagittal sinus, and repeatedly irrigate the drill bit with normal saline to prevent thermal injury to the cortex. After imaging the baseline conditions, apply a 10-millimeter segment of 2-0 silk suture soaked with 40%ferric chloride solution onto the exposed superior sagittal sinus surface for five minutes.Replace the suture with a newly soaked segment and apply for another five minutes. Then, rinse the surgical field with 0.5 milliliters of normal saline three times to remove residual ferric chloride. Close the skin using interrupted sutures with 4-0 silk suture, and disinfect the area with povidone-iodine.Observe the rat until it is fully awake, then return it to its cage. Turn on the laser speckle contrast imaging system and the corresponding software, RF LSI version five. Press the online mode button to activate live imaging.Adjust the height and position of the instrument until the indicator laser is centered within the field of view. Place the rat on a foam platform, and adjust its position until the superior sagittal sinus aligns with the indicator laser. Adjust the magnification and focus until the image is displayed clearly.Set the lower and upper limits of the pseudocolor threshold to optimize the visual appearance of the pseudocolor image. Set a circular ROI over the superior sagittal sinus, record venous flow, and save the original and pseudocolor images. Prepare isoflurane, ultrasound gel, disposable razor blades, medical tapes, and check all the instruments required.Install the three-dimensional acquisition motor onto the imaging system. Then, install the UHF 57X transducer, and initialize the motor. After anesthetizing the rat, place it in a prone position on the thermostatic imaging plate set to 37 degrees Celsius, and gently secure the head and limbs using medical tapes.Apply ultrasound gel evenly over the exposed scalp. Adjust the position and orientation of the rat relative to the transducer until the superior sagittal sinus and intraluminal thrombus are clearly visualized. Acquire sequential tomographic images of the superior sagittal sinus in the sagittal plane using three-dimensional imaging in B mode with a step size of 0.04 millimeters, followed by tomographic images in the coronal plane using the same settings.Use color Doppler mode to observe blood flow in the superior sagittal sinus, and apply pulsed-wave Doppler mode to measure blood flow velocity. Discontinue anesthesia and observe the rat until it is fully awake. Then, return the rat to its cage.Finally, export the acquired images, and analyze them using Vevo LAB software. Record the maximum sagittal and coronal cross-sectional areas and the volume of the thrombus in the superior sagittal sinus, along with the maximum blood flow velocity of the sinus. After local application of ferric chloride, laser speckle contrast imaging detected a significant reduction in local blood flow in the superior sagittal sinus compared with baseline.On postoperative day seven, small animal ultrasound confirmed thrombus formation in the superior sagittal sinus with partial luminal recanalization. Pulsed-wave Doppler ultrasound demonstrated measurable blood flow velocity within the residual lumen of the superior sagittal sinus. Three-dimensional ultrasound imaging enabled volumetric measurement of the superior sagittal sinus thrombus using serial image slices with a thickness of 0.04 millimeters.Our refined model is less invasive and more compatible with ultrasound imaging, enabling more reliable, direct therapeutic development and evaluation. Make sure that interoperative laser speckle contrast imaging confirms model's success and postoperative high-resolution ultrasound dynamically monitors thrombus formation, lysis, and lumen recanalization.

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Intracellular Ca2+ remodeling in aging may contribute to excitotoxicity and neuron damage, processes mediated by Ca2+ overload. We aimed at investigating Ca2+ remodeling in the aging brain using fluorescence and bioluminescence imaging of cytosolic and mitochondrial Ca2+ in long-term cultures of rat hippocampal neurons, a model of neuronal aging.

Fluorescence and Bioluminescence Imaging of Subcellular Ca2+ in Aged Hippocampal Neurons

This manuscript provides protocols that use in utero electroporation (IUE) to describe the structural connectivity of neurons at the single-cell level and the excitability of fluorescently labeled neurons. Histology is used to characterize dendritic and axonal projections. Whole-cell recording in acute slices is used to investigate excitability.

In Utero Electroporation Approaches to Study the Excitability of Neuronal Subpopulations and Single-cell Connectivity

Kinematic gait analysis in the sagittal plane yields highly precise information about how movement is executed. We describe the application of these techniques to identify gait deficits for mice subjected to autoimmune-mediated demyelination. These methods may also be used to characterize gait deficits for other mouse models featuring impaired locomotion.

Sagittal Plane Kinematic Gait Analysis in C57BL/6 Mice Subjected to MOG35-55 Induced Experimental Autoimmune Encephalomyelitis

Here, we present a protocol to introduce a rat model of central fatigue using the modified multiple platform method (MMPM).

A Rat Model of Central Fatigue Using a Modified Multiple Platform Method

Presented here is a protocol for establishing acute pontine infarction in a rat model via electrical stimulation with a single pulse.

Establishment of Acute Pontine Infarction in Rats by Electrical Stimulation

Learn a refined FeCl₃-soaked suture technique for inducing superior sagittal sinus thrombosis in rats with reduced cortical injury. This method enables reliable imaging and supports preclinical evaluation of cerebral venous thrombosis therapies.

Establishment of a Modified Ferric Chloride-Induced Superior Sagittal Sinus Thrombosis

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