Intravital Fluorescence Microscopy to Study Microvascular Thrombus Formation
Published: July 31, 2023
Abstract
Source: Strüder, D., et al. Intravital Microscopy and Thrombus Induction in the Earlobe of a Hairless Mouse. J. Vis. Exp. (2017)
This video demonstrates the intravital fluorescence microscopy technique to study thrombus formation in a mouse model. The mouse is injected with a fluorophore-labeled polysaccharide to monitor blood circulation. Upon illumination with a high-intensity light, the phototoxic induction of an injury in the microvasculature of the ear leads to the formation of a thrombus, which is confirmed by visualizing the obstruction in the blood flow.
Protocol
All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board. 1. Prearrangement of the Animals Weigh a mouse and load the respective drug (e.g., the cannabinoid, 5 mg/kg body weight (bw)) into an insulin syringe. Administer the drug 30 min prior to thrombus induction. By holding the neck of the mouse between the thumb and the index finger and the tail of the mouse …
Representative Results
Figure 1. Preparation of the Jugular Vein (A) and Placement of the Ear for Intravital Microscopy (B). (A) Using the operation stereomicroscope, the right jugular vein is prepared. The sutures for the placement of the left ear are stitched before the injection of FITC-dextran. The wound from the prior dissection of the left jugular vein is closed with transcutaneous sutures (B). The left ear is fixed with polypro…
Disclosures
The authors have nothing to disclose.
Materials
| SKH-1/hr mice | Charles River | 477 | Can be purchased from other vendors |
| Operation stereomicroscope | Leica | M651/M655 | Can be purchased from other vendors |
| Intravital microscope | Zeiss | Axiotech Vario 100 | Can be purchased from other vendors |
| Objective (20x/0.95) | Zeiss | 20x/0,50 W; Plan-NEOFLUAR | Can be purchased from other vendors |
| Objective (63x/0.95) | Zeiss | 63x/0,95 W; ACHROPLAN | Can be purchased from other vendors |
| Black and white CCD-camera | Pieper | FK 6990 IQ-S | Can be purchased from other vendors |
| DVD-recorder | Panasonic | DMR-EX99V | Can be purchased from other vendors |
| Sodium chloride | Braun | 5/12612055/1011 | Can be purchased from other vendors |
| Ketamine 10% | Bela pharm | F3901-6 | Can be purchased from other vendors |
| Xylazine 2% | Bayer | 6293841.00.00 | Can be purchased from other vendors |
| FITC-dextran 5% | Sigma | 46945-100MG-F | Can be purchased from other vendors |
| Dexapanthenol 5% eye ointment | Bayer | 6029009.00.00 | Can be purchased from other vendors |
| DMSO | Sigma | 472301 | Can be purchased from other vendors |
| Coverslips 5 x 5 x 1 mm | Menzel | L4339 | Can be purchased from other vendors |
| Plasters | Leukosilk | 4683400 | Can be purchased from other vendors |
| Cotton swabs | Sanyo | 604-A-1 | Can be purchased from other vendors |
| Infrared light | Beurer | 5/13855 | Can be purchased from other vendors |
| Single-use syringe | Braun | 2020-08 | Can be purchased from other vendors |
| Insulin syringe | Braun | 9161502 | Can be purchased from other vendors |
| Disposable hypodermic needles | Braun | 465 7640 | Can be purchased from other vendors |
| End-to-end capillary | Sarstedt | 19,447 | Can be purchased from other vendors |
| Heating plate | Klaus Effenberg | OP-T 185/03 | Can be purchased from other vendors |
| Scissors 14.5 cm | Aesculap | BC259R | Can be purchased from other vendors |
| Needle Holder | Aesculap | BM081R | Can be purchased from other vendors |
| Microforceps | Aesculap | BD331R | Can be purchased from other vendors |
| Microscissors | Aesculap | OC496R | Can be purchased from other vendors |
| Scalpel 21 | Dahlhausen | 11.000.00.511 | Can be purchased from other vendors |
| Prolene 7-0 | Ethicon | XNEH7470 | Can be purchased from other vendors |
| Electrocautery | Servoprax | H40140 | Can be purchased from other vendors |
| Acrylglass pad | Integrated heating, 0.5 cm high plane |
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Cite This Article
Intravital Fluorescence Microscopy to Study Microvascular Thrombus Formation. J. Vis. Exp. (Pending Publication), e21464, doi: (2023).