Objective: Induce focal cortical cerebral ischemia through electrocoagulation of the distal middle cerebral artery to create an animal stroke model
This is a Middle Cerebral Artery Occlusion by Electrocoagulation protocol using mouse as the model organism. The procedure involves 13 procedural steps, 5 equipment items, 4 materials. Extracted from a 2020 paper published in Nature Communications.
Model and subjects
mouse • C57BL/6J (PAD4 -/- knockout) and C57BL/6 (wild-type controls) • male • Age-matched • 23-26g
Study window
~15 minute study window | ~15 minutes hands-on
Core workflow
Animal housing and environmental conditions • Anesthesia induction • Surgical incision
Primary readouts
Key equipment and reagents
Verified items
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House mice in temperature-controlled environment with controlled light-dark cycle and humidity
Note: 12 h light-dark cycle with food and water available ad libitum; room humidity controlled at 55 ± 5%
“All mice were housed in a temperature-controlled environment (22 ± 2 °C) on a 12 h light–dark cycle with food and water available ad libitum. Room humidity was controlled at 55 ± 5%”
Anesthetize male mice weighing 23-26g with isoflurane in oxygen and nitrous oxide mixture
Note: Specific anesthesia induction time not mentioned
“Male mice weighing 23–26 g were anesthetized with 1–1.5% isoflurane in 30% oxygen and 70% nitrous oxide”
Make a 2 cm curved skin incision between the right eye and the right ear using surgical scissors
Note: Curved incision for optimal surgical access
“After making a 2 cm curved skin incision between the right eye and the right ear using surgical scissors”
Retract the temporal muscle laterally to expose the surgical field
“the temporal muscle was retracted laterally”
Under operating microscope, use high-speed micro drill to open a 1.5 mm-diameter window in the skull just rostral to the foramen ovale
Note: Window location is critical - just rostral to foramen ovale
“Under an operating microscope, a 1.5 mm-diameter window was opened using a high-speed micro drill (Stoelting, CellPoint Scientific, Maryland) just rostral to the foramen ovale”
Carefully open the dura mater to expose the distal middle cerebral artery
Note: Careful handling required to avoid damage to underlying tissue
“The dura mater was carefully opened and the distal MCA was exposed and isolated”
Expose, isolate, and electrocauterize the distal MCA using bipolar electrocoagulation forceps, then disconnect it just distal to crossing the olfactory tract
Note: Disconnection location is critical - just distal to olfactory tract crossing
“the distal MCA was exposed and isolated, electrocauterized using bipolar electrocoagulation forceps, and disconnected just distal to crossing the olfactory tract”
Immediately following MCA occlusion, occlude the right common carotid artery with a microvascular clip
Note: Must be done immediately after MCA occlusion; maintained for 15 minutes
“Immediately following the occlusion, the right common carotid artery (CCA) was occluded with a microvascular clip (Fine Science Tools, Foster City, CA) for 15 min”
Close surgical wounds and allow mice to recover from anesthesia
Note: Recovery conditions not specified
“After closing of the surgical wounds, mice were allowed to recover from the anesthesia”
For sham-operated control animals, expose the MCA and CCA but do not perform occlusion
Note: Sham controls undergo identical surgical procedure except for the occlusion step
“Sham-operated animals only had the MCA and the CCA exposed”
Prepare six coronal sections of 20 µm thickness from brain tissue
Note: Specific timing for tissue collection not mentioned
“For cerebral infarction, six coronal sections (20 µm) were stained with hematoxylin and eosin”
Stain coronal brain sections with hematoxylin and eosin
“six coronal sections (20 µm) were stained with hematoxylin and eosin”
Measure infarct volume using NIH ImageJ 1.46r software and present as percentage of contralateral hemisphere
Note: Infarct volume normalized to contralateral hemisphere for comparison
“Infarct volume was measured using the NIH ImageJ 1.46r software and was presented as percentage of the contralateral hemisphere”
This section explains what the experiment is doing, which readouts matter, what the data artifacts usually look like, and how the analysis should flow from raw capture to reported result.
Induce focal cortical cerebral ischemia through electrocoagulation of the distal middle cerebral artery to create an animal stroke model
Objective
Induce focal cortical cerebral ischemia through electrocoagulation of the distal middle cerebral artery to create an animal stroke model
Subjects
From papermouse • C57BL/6J (PAD4 -/- knockout) and C57BL/6 (wild-type controls) • male • Age-matched • 23-26g
Cohort notes
From paperPAD4 -/- mice purchased from The Jackson Laboratory; WT controls from SLAC Laboratory Animal Co.
Animal housing and environmental conditions
Anesthesia induction
Surgical incision
Muscle retraction
Cerebral infarct volume (measured as percentage of contralateral hemisphere)
From paperInfarct volume measured using NIH ImageJ 1.46r software and presented as percentage of the contralateral hemisphere
Artifact type
Representative image panels with region or marker comparisons
Comparison focus
Compare staining intensity, structure, or cell counts across matched conditions
Focal cortical ischemia induction success
From paperInfarct volume measured using NIH ImageJ 1.46r software and presented as percentage of the contralateral hemisphere
Artifact type
Representative image panels with region or marker comparisons
Comparison focus
Compare staining intensity, structure, or cell counts across matched conditions
Cerebral infarct volume (measured as percentage of contralateral hemisphere)
From paperRaw artifact
Field or section images captured from matched samples
Processed artifact
Selected representative panels with quantified intensity, counts, or area measurements
Final reported form
Per-group imaging summaries with representative figures and quantified endpoints
Focal cortical ischemia induction success
From paperRaw artifact
Field or section images captured from matched samples
Processed artifact
Selected representative panels with quantified intensity, counts, or area measurements
Final reported form
Per-group imaging summaries with representative figures and quantified endpoints
Acquisition
Capture matched images from the relevant tissue region using the same acquisition settings across samples.
Preprocessing / cleaning
Infarct volume measured using NIH ImageJ 1.46r software and presented as percentage of the contralateral hemisphere
Scoring or quantification
Quantify the primary readouts for this experiment: Cerebral infarct volume (measured as percentage of contralateral hemisphere); Focal cortical ischemia induction success.
Normalization
Normalize image-derived measurements against the matched acquisition or segmentation rules before comparing groups.
Statistical comparison
Statistical method not yet structured for this page.
Reporting output
Report representative outputs alongside summary comparisons for Cerebral infarct volume (measured as percentage of contralateral hemisphere), Focal cortical ischemia induction success.
Source links and direct wording from the methods section for validation and deeper review.
Citation
Lijing Kang et al. (2020). Neutrophil extracellular traps released by neutrophils impair revascularization and vascular remodeling after stroke. Nature Communications
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Stoelting, CellPoint Scientific
Fine Science Tools
National Institutes of Health
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Current status surfaces were computed from experiment data updated Mar 14, 2026.
Source access
Jump back into the original paper or the methods evidence section when you need exact wording, exclusions, or method-specific caveats.
This protocol has structured steps plus evidence quotes, and is ready for canonical sync.
Steps
13
Evidence Quotes
22
Protocol Items
9
Linked Products
0
Canonical Sync
Pending
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Computed from the current experiment record updated Mar 14, 2026.
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Steps
13
Evidence
22
Specific Products
0/0
Canonical Sync
Pending
What this score means
The verification score reflects evidence coverage, subject detail, paper provenance, step depth, and whether linked products resolve to specific item pages instead of generic searches.
Computed from the current experiment record updated Mar 14, 2026.
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