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In This Article

  • Summary
  • Abstract
  • Introduction
  • Protocol
  • Results
  • Discussion
  • Disclosures
  • Acknowledgements
  • Materials
  • References
  • Reprints and Permissions

Summary

Here, we describe a mouse model of retinal ischemia by transient bilateral common carotid artery occlusion using simple sutures and a clamp. This model can be useful for understanding the pathological mechanisms of retinal ischemia caused by cardiovascular abnormalities.

Abstract

Diverse vascular diseases such as diabetic retinopathy, occlusion of retinal veins or arteries and ocular ischemic syndrome can lead to retinal ischemia. To investigate pathological mechanisms of retinal ischemia, relevant experimental models need to be developed. Anatomically, a main retinal blood supplying vessel is the ophthalmic artery (OpA) and OpA originates from the internal carotid artery of the common carotid artery (CCA). Thus, disruption of CCA could effectively cause retinal ischemia. Here, we established a mouse model of retinal ischemia by transient bilateral common carotid artery occlusion (tBCCAO) to tie the right CCA with 6-0 silk sutures and to occlude the left CCA transiently for 2 seconds via a clamp, and showed that tBCCAO could induce acute retinal ischemia leading to retinal dysfunction. The current method reduces reliance on surgical instruments by only using surgical needles and a clamp, shortens occlusion time to minimize unexpected animal death, which is often seen in mouse models of middle cerebral artery occlusion, and maintains reproducibility of common retinal ischemic findings. The model can be utilized to investigate the pathophysiology of ischemic retinopathies in mice and further can be used for in vivo drug screening.

Introduction

The retina is a neurosensory tissue for visual function. Since a substantial amount of oxygen is needed for visual function, the retina is known as one of the highest oxygen demanding tissues in the body1. The retina is susceptible to vascular diseases as oxygen is delivered through blood vessels. Various types of vascular diseases, such as diabetic retinopathy and retinal blood vessel (veins or arteries) occlusion, can induce retinal ischemia. To investigate pathological mechanisms of retinal ischemia, reproducible and clinically relevant experimental models of retinal ischemia are considered necessary. Middle cerebral artery occlusion (MCAO) by insertion of an intraluminal filament is the most generally utilized method for the development of in vivo rodent models of experimental cerebral ischemia2,3. Due to the proximity of the ophthalmic artery (OpA) to MCA, MCAO models are also used simultaneously to understand the pathophysiology of retinal ischemia4,5,6. To induce cerebral ischemia along with retinal ischemia, long filaments are typically inserted through incision of the common carotid artery (CCA) or the external carotid artery (ECA). These methods are difficult to perform, require a long time to complete the surgery (over 60 minutes for one mouse), and lead to high variabilities in the outcomes after the surgery7. It remains important to develop a better model to improve these concerns.

In this study, we simply used short transient bilateral CCA occlusion (tBCCAO) with needles and a clamp to induce retinal ischemia in mice and analyzed typical results of ischemic injuries in the retina. In this video, we will give a demonstration of the tBCCAO procedure.

Protocol

All methods described here have been approved by the Institutional Animal Care and Use Committee (IACUC) of Keio University School of Medicine.

1. Preparation of surgical instruments and animals

  1. Autoclave surgical instruments and keep them in 70% ethyl alcohol. Prior to each new surgical procedure, clean surgical instruments carefully using 70% ethyl alcohol.
  2. Prepare male BALB/cAJc1 mice (6 weeks old, 26-28 kg) in a specific-pathogen-free (SPF) room to maintain sterile conditions before, during and after the surgery.

2. Transient bilateral common carotid artery occlusion (tBCCAO)

  1. Put a mouse under anesthesia via intraperitoneal injection with a combination of midazolam (40 μg/100 μL), medetomidine (7.5 μg/100 μL) and butorphanol tartrate (50 μg/100 μL), termed “MMB”, as previously described8,9. Hold the mouse’s back skins to keep the mouse away from bumping its eyes until the mouse is completely anesthetized.
    1. Judge the depth of anesthesia by pinching the mouse toe until it has no response, of which method is commonly used for checking complete anesthesia10.
      NOTE: Generally, less than 5 min are required for mice to fall asleep. Proper recipes for general anesthesia may be different by institutions.
  2. Apply one drop of 0.1% purified sodium hyaluronate eye drop solution to the eyes to prevent dryness on the eyes under anesthesia.
  3. Place the mouse on its back and fix the mouse's paws using adhesive tapes.
  4. Disinfect the neck area of the mouse using 70% ethyl alcohol before the surgery.
    NOTE: Additional clipping of the fur was not performed as this may cause subsequent skin inflammation11,12.
  5. Perform sagittal incision of the neck with a blade (Figure 1).
    NOTE: Incision needs to be made on the midline between the neck, sternum and trachea.
  6. Separate both salivary glands carefully using two forceps and mobilize them to visualize the underlying CCAs.
  7. Isolate the right CCA carefully from the respective vagal nerves and accompanying veins without harming their structures, and place two 6-0 silk sutures under the CCA. Tie the two ties tightly to block the blood flow (Figure 1).
    NOTE: During the procedure, small veins could be damaged. If bleeding is seen, wiping is required to visualize the CCAs clearly.
  8. Find the left CCA carefully from the respective vagal nerves and accompanying veins without harming their structures, and occlude the left CCA for 2 seconds by a clamp (Figure 1).
    NOTE: A 6-0 silk suture needle is needed to be placed under the left CCA to mark a site for clamping.
  9. After reopening of the left CCA, suture wounds of the neck by a 6-0 silk suture and apply a dab of antibiotic (50 μL) onto the neck to inhibit bacterial infection.
    NOTE: Softly remove a clamp to avoid damaging the arterial wall when reopening of the left CCA.
  10. Inject 0.75 mg/kg of atipamezole hydrochloride intraperitoneally to the mouse to help the mouse recovered from deep anesthesia quickly. Return the mouse to a mouse cage with pre-heated pads.
    NOTE: Do not let the mouse left unattended until the mouse regains sufficient consciousness to maintain sternal recumbency.
  11. Inject 0.4 mg/kg of butorphanol tartrate to the mouse for the management of pain when the mouse wakes up.
    NOTE: The protocol can be paused here. As a first hint for successful tBCCAO, eyelid drooping of the mouse can be observed (Figure 2).
  12. For euthanasia, inject 3x of MMB mixture to the mice and sacrifice them for experiments.

3. General observations (survival rates and eyelid drooping)

  1. After the surgery, check survival rates for all causes of death at day 0 (after the surgery), 1, 3 and 7.
  2. Assess eyelid drooping by a 4-point rating scale: 1 = no drooping, 2 = mild drooping (~50%), 3 = severe drooping (over 50%), and 4 = severe drooping with eye discharge.

4. Retinal blood perfusion

  1. Inject 200 μL of FITC-dextran (25 mg/mL) into the left ventricle of the mouse, which is commonly used for the observation of blood perfusion in mouse retinal vessels13,14.
  2. 2 minutes after circulation, enucleate the eyes and fix in 4% paraformaldehyde for 1 hour. The retinas were carefully obtained and flat-mounted, as previously described15, and examined via a fluorescence microscope.
  3. Take photographs of the retinal whole mounts at 4x magnification and merge into a single using a merge analyzer, previously described16.
  4. Measure the perfused areas via a vessel analysis tool in NIH Fiji/ImageJ software.

5. Western blot

  1. 3 and 6 hours after tBCCAO, obtain the eyes of mice and immediately transfer to a Petri dish containing cold PBS to isolate the retinas.
  2. After isolation of the retinas, perform western blotting, as previously described9.
  3. Incubate with antibodies for hypoxia-inducible factor-1α (HIF-1α; a general hypoxia marker) and for β-Actin (an internal loading control) overnight followed by incubation of HRP-conjugated secondary antibodies. Visualize the signals via chemiluminescence.

6. Quantitative PCR (qPCR)

  1. 6, 12 and 24 hours after tBCCAO, process the obtained retinas for qPCR, as previously described17.
  2. Perform qPCR via real-time PCR system. Primers used are listed in Table 1. Calculate fold changes between levels of different transcripts by the ΔΔCT method.

7. Immunohistochemistry (IHC)

  1. 3 days after tBCCAO, obtain the eyes of mice and embed in paraffin.
  2. Cut the paraffin-embedded eyes by a microtome to obtain the eye sections.
  3. De-paraffinize and stain the eye sections of 5 μm thickness as previously described13.
  4. Incubate with an antibody for glial fibrillary acidic protein (GFAP; a reliable marker for astrocytes and Müller cells in the retina) overnight followed by incubation of Alexa Fluor 555-conjugated secondary antibody.
  5. Use DAPI (4′,6-diamidino-2-phenylindole) for staining the nucleus in the retina. Visualize signals via a fluorescence microscope.
  6. Assess morphology scoring by a 4-point rating scale, as previously described13,18: 0 = no signal, 1 = few positive glial end-feet in the ganglion cell layer (GCL), 2 = few labelled processes reaching from GCL to the outer nuclear layer (ONL), and 3 = most labelled processes reaching from GCL to ONL.

8. Electroretinography (ERG)

  1. 3 and 7 days after tBCCAO, perform ERG using a Ganzfeld dome, acquisition system and LED stimulators, as previously described9.
  2. Following dark adaptation overnight, anesthetize mice with a combination of MMB under dim red light.
  3. Use a mixed solution of 0.5% tropicamide and 0.5% phenylephrine to dilate the pupils.
  4. Place the active electrodes on contact lens and place the reference electrode in the mouth.
  5. Obtain ERG responses from both eyes of each animal.
  6. Record scotopic responses under dark adaptation with various stimuli.
  7. Measure the amplitudes of a-wave from the baseline to the lowest point of a-wave.
  8. Measure the amplitudes of b-wave from the lowest point of a-wave to the peak of b-wave.
  9. Keep all mice warm during the procedure using heat pads.

9. Optical coherence tomography (OCT)

  1. 2 weeks after tBCCAO, perform OCT using SD-OCT system, as previously reported8,9.
  2. For the measurement, subject mice to mydriasis by a mixed solution of 0.5% tropicamide and 0.5% phenylephrine, and to general anesthesia by a mixture of MMB.
  3. Obtain B scan images from equatorial slices of en-face scans.
  4. Examine the retinas at 0.2, 0.4 and 0.6 mm from the optic nerve head.
  5. Measure retinal thickness from the retinal nerve fiber layer (NFL) to the external limiting membrane (ELM), and consider the average of measured values as retinal thickness of an individual mouse.
  6. Plot the results as spider diagrams.

Results

After systemic circulation of FITC-dextran for 2 minutes, retinal vasculatures of the left and right retinas in the sham-operated mice and tBCCAO-operated mice were examined (Supplementary Figure 1). FITC-dextran was fully visible in the both retinas in the sham-operated mice and the left retina in the tBCCAO-operated mice, while it was partially detectible in the right retina in the tBCCAO-operated mice.

After tBCCAO, eyelid drooping was examined (Figure ...

Discussion

In the study, we have shown that tBCCAO, using simple sutures and a clamp, could induce retinal ischemia and accompanying retinal dysfunction. Furthermore, we have demonstrated our current protocol for the development of a mouse model of retinal ischemia is easier and faster in comparison with other previous protocols for the development of retinal ischemic injury models2,3,7.

Anatomically, the left a...

Disclosures

The authors have nothing to disclose.

Acknowledgements

This work was supported by Grants-in-Aid for Scientific Research (KAKENHI) (18K09424 to Toshihide Kurihara and 20K18393 to Yukihiro Miwa) from the Ministry of Education, Culture, Sports, Science and Technology (MEXT).

Materials

NameCompanyCatalog NumberComments
Atipamezole hydrochlorideZenoaqAntisedanFor anti-anesthesia
Applied Biosystems 7500 FastApplied Biosystems-For qPCR
Butorphanol tartrateMeiji Seika PharmaVetorphaleFor anesthesia
BZ-II AnalyzerKEYENCE-For an image merge
BALB/cAJc1CLEA-Mouse strain
β-Actin (8H10D10) Mouse mAbCST3700For western blot
Clamp ForcepWorld Precision InstrumentsWPI 500451For surgery
Dumont forceps #5Fine Science Tools11251-10For surgery
DAPI solutionDojindo340-07971For IHC
Envisu SD-OCT systemLeicaR4310For OCT
FITC-dextranMerkFD2000SFor retinal blood perfusion
Fluorescence microscopeKEYENCEBZ-9000For fluorescence detection
Gatifloxacin hydrateSenju PharmaceuticalGachifuroFor anti-bacterial infection
GFAP Monoclonal Antibody (2.2B10)Thermo13-0300For IHC
Heating padMarukanRH-200For surgery
HIF-1α (D1S7W) XP Rabbit mAbCST36169For western blot
ImageQuant LAS 4000 miniGE Healthcare-For chemiluminescence
MidazolamSandoz K.KSANDOZFor anesthesia
Microtome Tissue-Tek TEC 6Sakura-For sectioning
MedetomidineOrion CorporationDomitorFor anesthesia
Needle holderHandayaHS-2307For surgery
PuRECMAYO Corporation-For ERG
ScissorFine Science Tools91460-11For surgery
Sodium hyaluronateSanten PharmaceuticalHyaleinFor eye lubrication
Tropicamide/Penylephrine hydrochlorideSanten PharmaceuticalMydrin-PFor mydriasis
6-0 silk sutureNatsumeE12-60N2For surgery

References

  1. Anderson, B. Ocular effects of changes in oxygen and carbon dioxide tension. Transactions of the American Ophthalmological Society. 66, 423-474 (1968).
  2. Ingberg, E., Dock, H., Theodorsson, E., Theodorsson, A., Ström, J. O. Method parameters' impact on mortality and variability in mouse stroke experiments: a meta-analysis. Scientific Reports. 6 (1), 21086 (2016).
  3. Atochin, D. N., Clark, J., Demchenko, I. T., Moskowitz, M. A., Huang, P. L. Rapid Cerebral Ischemic Preconditioning in Mice Deficient in Endothelial and Neuronal Nitric Oxide Synthases. Stroke. 34 (5), 1299-1303 (2003).
  4. Allen, R. S., et al. Severity of middle cerebral artery occlusion determines retinal deficits in rats. Experimental Neurology. 254, 206-215 (2014).
  5. Steele, E. C., Guo, Q., Namura, S. Filamentous Middle Cerebral Artery Occlusion Causes Ischemic Damage to the Retina in Mice. Stroke. 39 (7), 2099-2104 (2008).
  6. Minhas, G., Morishita, R., Anand, A. Preclinical models to investigate retinal ischemia: advances and drawbacks. Frontiers in Neurology. 3, 75 (2012).
  7. McColl, B. W., Carswell, H. V., McCulloch, J., Horsburgh, K. Extension of cerebral hypoperfusion and ischaemic pathology beyond MCA territory after intraluminal filament occlusion in C57Bl/6J mice. Brain Res. 997 (1), 15-23 (2004).
  8. Jiang, A. X., et al. Inducement and Evaluation of a Murine Model of Experimental Myopia. Journal of Visualized Experiments. (143), e58822 (2019).
  9. Miwa, Y., et al. Pharmacological HIF inhibition prevents retinal neovascularization with improved visual function in a murine oxygen-induced retinopathy model. Neurochemistry International. 128, 21-31 (2019).
  10. Adams, S., Pacharinsak, C. Mouse Anesthesia and Analgesia. Current Protocols in Mouse Biology. 5 (1), 51-63 (2015).
  11. Speetzen, L. J., Endres, M., Kunz, A. Bilateral Common Carotid Artery Occlusion as an Adequate Preconditioning Stimulus to Induce Early Ischemic Tolerance to Focal Cerebral Ischemia. Journal of Visualized Experiments. (75), e4387 (2013).
  12. Engel, O., Kolodziej, S., Dirnagl, U., Prinz, V. Modeling stroke in mice - middle cerebral artery occlusion with the filament model. Journal of Visualized Experiments. (47), e2423 (2011).
  13. Lee, D., Kang, H., Yoon, K. Y., Chang, Y. Y., Song, H. B. A mouse model of retinal hypoperfusion injury induced by unilateral common carotid artery occlusion. Experimental Eye Research. 201, 108275 (2020).
  14. Li, S., et al. Retro-orbital injection of FITC-dextran is an effective and economical method for observing mouse retinal vessels. Molecular Vision. 17, 3566-3573 (2011).
  15. Tual-Chalot, S., Allinson, K. R., Fruttiger, M., Arthur, H. M. Whole Mount Immunofluorescent Staining of the Neonatal Mouse Retina to Investigate Angiogenesis In vivo. Journal of Visualized Experiments. (77), e50546 (2013).
  16. Lee, D., et al. A Fairy Chemical Suppresses Retinal Angiogenesis as a HIF Inhibitor. Biomolecules. 10 (10), (2020).
  17. Tomita, Y., et al. Pemafibrate Prevents Retinal Pathological Neovascularization by Increasing FGF21 Level in a Murine Oxygen-Induced Retinopathy Model. International Journal of Molecular Sciences. 20 (23), 5878 (2019).
  18. Yamamoto, H., Schmidt-Kastner, R., Hamasaki, D. I., Yamamoto, H., Parel, J. M. Complex neurodegeneration in retina following moderate ischemia induced by bilateral common carotid artery occlusion in Wistar rats. Experimental Eye Research. 82 (5), 767-779 (2006).
  19. Cheng, L., Yu, H., Yan, N., Lai, K., Xiang, M. Hypoxia-Inducible Factor-1α Target Genes Contribute to Retinal Neuroprotection. Frontiers in Cellular Neuroscience. 11, 20 (2017).
  20. Mole, D. R., et al. Genome-wide association of hypoxia-inducible factor (HIF)-1alpha and HIF-2alpha DNA binding with expression profiling of hypoxia-inducible transcripts. The Journal of Biological Chemistry. 284 (25), 16767-16775 (2009).
  21. Majmundar, A. J., Wong, W. J., Simon, M. C. Hypoxia-Inducible Factors and the Response to Hypoxic Stress. Molecular Cell. 40 (2), 294-309 (2010).
  22. Newman, E. A. Glial cell regulation of neuronal activity and blood flow in the retina by release of gliotransmitters. Philosophical Transactions of the Royal Society B: Biological Sciences. 370 (1672), (2015).
  23. Vecino, E., Rodriguez, F. D., Ruzafa, N., Pereiro, X., Sharma, S. C. Glia-neuron interactions in the mammalian retina. Progress in Retinal and Eye Research. 51, 1-40 (2016).
  24. Symonds, C. The Circle of Willis. British Medical Journal. 1 (4906), 119 (1955).
  25. Lo, W. B., Ellis, H. The circle before willis: a historical account of the intracranial anastomosis. Neurosurgery. 66 (1), 7-18 (2010).
  26. Yang, G., et al. C57BL/6 strain is most susceptible to cerebral ischemia following bilateral common carotid occlusion among seven mouse strains: selective neuronal death in the murine transient forebrain ischemia. Brain Research. 752 (1), 209-218 (1997).
  27. Farkas, E., Luiten, P. G. M., Bari, F. Permanent, bilateral common carotid artery occlusion in the rat: A model for chronic cerebral hypoperfusion-related neurodegenerative diseases. Brain Research Reviews. 54 (1), 162-180 (2007).
  28. Morris, G. P., et al. A Comparative Study of Variables Influencing Ischemic Injury in the Longa and Koizumi Methods of Intraluminal Filament Middle Cerebral Artery Occlusion in Mice. PLOS ONE. 11 (2), 0148503 (2016).
  29. Tsuchiya, D., Hong, S., Kayama, T., Panter, S. S., Weinstein, P. R. Effect of suture size and carotid clip application upon blood flow and infarct volume after permanent and temporary middle cerebral artery occlusion in mice. Brain Research. 970 (1-2), 131-139 (2003).
  30. Kaelin, W. G., Ratcliffe, P. J. Oxygen Sensing by Metazoans: The Central Role of the HIF Hydroxylase Pathway. Molecular Cell. 30 (4), 393-402 (2008).
  31. Pauly, M., Sruthi, R. Ptosis: evaluation and management. Kerala Journal of Ophthalmolgy. 31 (1), 11-16 (2019).
  32. Averbuch-Heller, L., Leigh, R. J., Mermelstein, V., Zagalsky, L., Streifler, J. Y. Ptosis in patients with hemispheric strokes. Neurology. 58 (4), 620 (2002).
  33. Dutton, J. . Atlas of clinical and surgical orbital anatomy, second edition. 113, 1364 (2011).
  34. Ritzel, R. M., et al. Early retinal inflammatory biomarkers in the middle cerebral artery occlusion model of ischemic stroke. Molecular Vision. 22, 575-588 (2016).
  35. Crespo-Garcia, S., et al. Individual and temporal variability of the retina after chronic bilateral common carotid artery occlusion (BCCAO). PLOS ONE. 13 (3), 0193961 (2018).
  36. Qin, Y., et al. Functional and morphologic study of retinal hypoperfusion injury induced by bilateral common carotid artery occlusion in rats. Scientific Reports. 9 (1), 80 (2019).
  37. Block, F., Grommes, C., Kosinski, C., Schmidt, W., Schwarz, M. Retinal ischemia induced by the intraluminal suture method in rats. Neuroscience Letters. 232 (1), 45-48 (1997).
  38. Allen, R. S., et al. Progesterone Treatment in Two Rat Models of Ocular Ischemia. Investigative Ophthalmology & Visual Science. 56 (5), 2880-2891 (2015).
  39. Miller, R. F., Dowling, J. E. Intracellular responses of the Müller (glial) cells of mudpuppy retina: their relation to b-wave of the electroretinogram. Journal of Neurophysiology. 33 (3), 323-341 (1970).
  40. Block, F., Grommes, C., Kosinski, C., Schmidt, W., Schwarz, M. Retinal ischemia induced by the intraluminal suture method in rats. Neuroscience Letters. 232 (1), 45-48 (1997).
  41. Lee, J. H., Shin, J. M., Shin, Y. J., Chun, M. H., Oh, S. J. Immunochemical changes of calbindin, calretinin and SMI32 in ischemic retinas induced by increase of intraocular pressure and by middle cerebral artery occlusion. Anatomy & Cell Biology. 44 (1), 25-34 (2011).
  42. Li, S. Y., et al. Lycium barbarum polysaccharides reduce neuronal damage, blood-retinal barrier disruption and oxidative stress in retinal ischemia/reperfusion injury. PLOS ONE. 6 (1), 16380 (2011).
  43. Furashova, O., Matthé, E. Retinal Changes in Different Grades of Retinal Artery Occlusion: An Optical Coherence Tomography Study. Investigative Ophthalmology & Visual Science. 58 (12), 5209-5216 (2017).
  44. Zadeh, J. K., et al. Short-Time Ocular Ischemia Induces Vascular Endothelial Dysfunction and Ganglion Cell Loss in the Pig Retina. International Journal of Molecular Sciences. 20 (19), (2019).
  45. Liu, S., Zhen, G., Meloni, B. P., Campbell, K., Winn, H. R. Rodent stroke model guidelines for preclinical stroke trials (1st edition). Journal of Experimental Stroke & Translational Medicine. 2 (2), 2-27 (2009).
  46. Tang, Y., et al. Hypothermia-induced ischemic tolerance is associated with Drp1 inhibition in cerebral ischemia-reperfusion injury of mice. Brain Research. 1646, 73-83 (2016).
  47. Barone, F. C., Knudsen, D. J., Nelson, A. H., Feuerstein, G. Z., Willette, R. N. Mouse strain differences in susceptibility to cerebral ischemia are related to cerebral vascular anatomy. Journal of Cerebral Blood Flow & Metabolism. 13 (4), 683-692 (1993).
  48. Pula, J. H., Yuen, C. A. Eyes and stroke: the visual aspects of cerebrovascular disease. Stroke and Vascular Neurology. 2 (4), 210 (2017).
  49. Steele, E. C., Guo, Q., Namura, S. Filamentous middle cerebral artery occlusion causes ischemic damage to the retina in mice. Stroke. 39 (7), 2099-2104 (2008).
  50. Sim, D. A., et al. The Effects of Macular Ischemia on Visual Acuity in Diabetic Retinopathy. Investigative Ophthalmology & Visual Science. 54 (3), 2353-2360 (2013).
  51. Wu, K. K., Huan, Y. Streptozotocin-induced diabetic models in mice and rats. Current Protocols in Pharmacology. , (2008).
  52. Mubarak, A., Hodgson, J. M., Considine, M. J., Croft, K. D., Matthews, V. B. Supplementation of a high-fat diet with chlorogenic acid is associated with insulin resistance and hepatic lipid accumulation in mice. Journal of Agricultural and Food Chemistry. 61 (18), 4371-4378 (2013).
  53. Ansari, S., Azari, H., McConnell, D. J., Afzal, A., Mocco, J. Intraluminal middle cerebral artery occlusion (MCAO) model for ischemic stroke with laser doppler flowmetry guidance in mice. Journal of Visualized Experiments. (51), e2879 (2011).
  54. Hedna, V. S., et al. Validity of Laser Doppler Flowmetry in Predicting Outcome in Murine Intraluminal Middle Cerebral Artery Occlusion Stroke. Journal of Vascular and Interventional Neurology. 8 (3), 74-82 (2015).

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