Subject Area:	Medicine and Dentistry
More specific subject area:	Microvascular analysis of noise induced inflammation and microvascular dysfunction
Method name:	In vivo fluorescence microscopy and cerebral arteriole cannulation to assess noise induced changes in activation of white blood cells and microvascular dysfunction
Name and reference of original method:	In vivo microscopy of dorsal skinfold chambers •Algire GH, Legallais FY. Recent developments in the transparent-chamber technique as adapted to the mouse. J Natl Cancer Inst. 1949;10(2):225-53, incl 8 pl. Epub 1949/10/01. PubMed PMID:15393709. •Sandison JC. A new method for the microscopic study of living growing tissues by the introduction of a transparent chamber in the rabbit's ear. The Anatomical Record. 1924;28(4):281-7. doi: https://doi.org/10.1002/ar.1090280403. •Endrich B, Asaishi K, Gotz A, Messmer K. Technical report–a new chamber technique for microvascular studies in unanesthetized hamsters. Res Exp Med (Berl). 1980;177(2):125-34. Epub 1980/01/01. doi: 10.1007/BF01851841. PubMed PMID: 7003665. Cerebral arteriole cannulation •Reference of original method is not available. It was first described by our working group: Eckrich J, Frenis K, Rodriguez-Blanco G, Ruan Y, Jiang S, Bayo Jimenez MT, et al. Aircraft noise exposure drives the activation of white blood cells and induces microvascular dysfunction in mice. Redox Biol. 2021;46:102063. Epub 2021/07/19. doi: 10.1016/j.redox.2021.102063. PubMed PMID:34274810.
Resource availability:	Dorsal skinfold chamber preparation Titanium frame of dorsal skinfold chamber (custom made) Autoclave (Tuttnauer systec 5050 ELVD, Tuttnauer Europe B.V., Breda, Netherlands) Ketamine (Ketanest; Pfizer Pharma GmbH, Berlin, Germany) Xylazine (Rompun; Bayer, Leverkusen, Germany) NaCl 0.9% (B.Braun 0,9% Mini-Plasco® connect, B. Braun Melsungen AG, Melsungen, Germany) Heated operation platform (MEDAX GmbH & Co KG, Neumünster, Germany) Eye ointment (Corneregel®, Bausch & Lomb Inc., Rochester, New York, USA; Bepathen® Eye and Nose Ointment, Bayer AG, Leverkusen, Germany) Electric razor (Remington Contour, Spectrum Brands, Middleton, Wisconsin, USA) Depilatory cream (Veet, Reckitt Benckiser Deutschland GmbH, Heidelberg, Germany) Disinfectant spray (octenisept spray, Schülke & Mayr GmbH, Norderstedt, Germany) Surgical sutures (ETHIBOND EXCEL V5 - USP 4-0, 0.75m, Ethicon Inc., Raritan, New Jersey, USA) 1,5 mm Strauss cannula (DISPOMED GmbH & Co. KG, Gelnhausen, Germany) surgical pen (Securline Skin Marker, Aspen Surgical Products, Caledonia, Michigan, USA). Surgical microscope (ZEISS OPMI 1 FC, Carl Zeiss Meditec AG, Oberkochen, Germany) Cover glass slip (11.8 mm, Hecht Assistent, Glaswarenfabrik Karl Hecht GmbH & Co KG, Sondheim, Germany) Analgesia (Tramadol-ratiopharm, ratiopharm GmbH, Ulm, Germany) In vivo microscopy Fluorescein isothiocyanate (FITC)-labelled dextran (average mol wt 500 000, Sigma, Deisenhofen, Germany) Rhodamine 6G (Sigma, Deisenhofen, Germany) Local anesthesia (Emla® 25 mg/g Lidocain +25 mg/g Prilocain creme, Aspen Germany GmbH, Munich, Germany)
	Surgical canulla( 0.3 × 13mm, BD Microlance 3, Becton, Dickinson and Company Ltd., Drogheda, Ireland) Intravital microscope (Olympus BXFM, Olympus Deutschland GmbH, Hamburg, Germany) Cell Sens Dimension (Olympus Deutschland GmbH, Hamburg, Germany) Off-line computer-aided videoframe analysis (Cap Image, Dr. Zeintl Ingenieurbüro, Dreieich, Germany) Mechanical hand counter (Roth Selection, Carl Roth GmbH + Co. KG, Karlsruhe, Germany) Cerebral arteriole studies Precision tweezers (type 5, straight with extra fine tips, LH53.1, Carl Roth GmbH + Co. KG, Karlsruhe, Germany) Vannas capsulotomy scissors (straight, G-19760, Geuder AG, Heidelberg, Germany) Glass Capillaries (1.2 × 0.8 mm outer/inner diameter, 9-000-1211, Drummond Scientific Company, Broomall, PA, USA) Nylon Suture (10-0, 198001, Alcon AG, Freiburg, Switzerland) Vertical Pipette Puller (Model 700C, David Kopf Instruments, Tujunga, CA, USA) Pressure Myograph (Jim's Instruments Manufacturing Inc., Iowa City, IA, USA) Pericyclic pump (Model CYCLO II, EP76.1, Carl Roth GmbH + Co. KG, Karlsruhe, Germany) Recirculating water bath (Model FE 2, Thermo Haake GmbH, Karlsruhe, Germany) Inverted microscope (Model SXB-1A, Lowest Price Microscopes, MO, USA) Digital camera (Model TK-C1381, JVC Deutschland GmbH, Bad Vilbel, Germany) Calcium chloride (5239.1, Carl Roth GmbH + Co. KG, Karlsruhe, Germany) Kalium chloride (6781.1, Carl Roth GmbH + Co. KG, Karlsruhe, Germany) Kalium dihydrogen phosphate (3904.2, Carl Roth GmbH + Co. KG, Karlsruhe, Germany) Magnesium sulphate (261.2, Carl Roth GmbH + Co. KG, Karlsruhe, Germany) Sodium chloride (9265.2, Carl Roth GmbH + Co. KG, Karlsruhe, Germany) Sodium hydrogen carbonate (0965.3, Carl Roth GmbH + Co. KG, Karlsruhe, Germany) α-(D)-(+)- Glucose monohydrate (6780.1, Carl Roth GmbH + Co. KG, Karlsruhe, Germany) 9,11-dideoxy-9α,11α-methanoepoxy prostaglandin F2α (U-46619, 16450, Cayman Chemical, Ann Arbor, MI, USA) Acetylcholine chloride (A6625-25G, Merck KGaA, Darmstadt, Germany) Sodium nitroprusside (1065410025, Merck KGaA, Darmstadt, Germany)