12. Parker RI, Rick ME, Gralnick HR (1984) A method to minimize platelet activation during platelet isolation. Thromb Res 36: 265–270. 13. Celi A, Merrill-Skoloff G, Gross P, Falati S, Sim DS, et al. (2003) Thrombus formation: direct real-time observation and digital analysis of thrombus assembly in a living mouse by confocal and widefield intravital microscopy. J Thromb Haemost 1: 60–68. 14. Nieswandt B, Echtenacher B, Wachs FP, Schroder J, Gessner JE, et al. (1999) Acute systemic reaction and lung alterations induced by an antiplatelet integrin gpIIb/IIIa antibody in mice. Blood 94: 684–693. 15. Falati S, Gross P, Merrill-Skoloff G, Furie BC, Furie B (2002) Real-time in vivo imaging of platelets, tissue factor and fibrin during arterial thrombus formation in the mouse. Nat Med 8: 1175–1181. 16. Jasuja R, Furie B, Furie BC (2010) Endothelium-derived but not platelet-derived protein disulfide isomerase is required for thrombus formation in vivo. Blood 116: 4665–4674. 17. Larson MK, Watson SP (2006) Regulation of proplatelet formation and platelet release by integrin alpha IIb beta3. Blood 108: 1509–1514. 18. Weber C, Springer TA (1997) Neutrophil accumulation on activated, surface- adherent platelets in flow is mediated by interaction of Mac-1 with fibrinogen bound to alphaIIbbeta3 and stimulated by platelet-activating factor. J Clin Invest 100: 2085–2093. 19. Hechler B, Nonne C, Eckly A, Magnenat S, Rinckel JY, et al. (2010) Arterial thrombosis: relevance of a model with two levels of severity assessed by histologic, ultrastructural and functional characterization. J Thromb Haemost 8: 173–184. 20. Perrault C, Mangin P, Santer M, Baas MJ, Moog S, et al. (2003) Role of the intracellular domains of GPIb in controlling the adhesive properties of the platelet GPIb/V/IX complex. Blood 101: 3477–3484. 21. Petri B, Broermann A, Li H, Khandoga AG, Zarbock A, et al. (2010) von Willebrand factor promotes leukocyte extravasation. Blood 116: 4712–4719. 22. Zhang J, Varas F, Stadtfeld M, Heck S, Faust N, et al. (2007) CD41-YFP mice allow in vivo labeling of megakaryocytic cells and reveal a subset of platelets hyperreactive to thrombin stimulation. Exp Hematol 35: 490–499. 23. Arase H, Saito T, Phillips JH, Lanier LL (2001) Cutting edge: the mouse NK cell-associated antigen recognized by DX5 monoclonal antibody is CD49b (alpha 2 integrin, very late antigen-2). J Immunol 167: 1141–1144. 24. Pellicci DG, Hammond KJ, Coquet J, Kyparissoudis K, Brooks AG, et al. (2005) DX5/CD49b-positive T cells are not synonymous with CD1d-dependent NKT cells. J Immunol 175: 4416–4425. 25. Kehrel B (1995) Platelet receptors for collagens. Platelets 6: 11–16. 26. Hedrich H (2004) The Laboratory Mouse. San Francisco, USA: Elsevier Academic Press. 277 p. 27. Buttrum SM, Hatton R, Nash GB (1993) Selectin-mediated rolling of neutrophils on immobilized platelets. Blood 82: 1165–1174. 28. Diacovo TG, Roth SJ, Buccola JM, Bainton DF, Springer TA (1996) Neutrophil rolling, arrest, and transmigration across activated, surface-adherent platelets via sequential action of P-selectin and the beta 2-integrin CD11b/CD18. Blood 88: 146–157. 29. Palabrica T, Lobb R, Furie BC, Aronovitz M, Benjamin C, et al. (1992) Leukocyte accumulation promoting fibrin deposition is mediated in vivo by P- selectin on adherent platelets. Nature 359: 848–851. 30. Zarbock A, Polanowska-Grabowska RK, Ley K (2007) Platelet-neutrophil- interactions: linking hemostasis and inflammation. Blood Rev 21: 99–111. 31. Maugeri N, Rovere-Querini P, Evangelista V, Covino C, Capobianco A, et al. (2009) Neutrophils phagocytose activated platelets in vivo: a phosphatidylserine, P-selectin, and {beta}2 integrin-dependent cell clearance program. Blood 113: 5254–5265. 32. Kornerup KN, Salmon GP, Pitchford SC, Liu WL, Page CP (2010) Circulating platelet-neutrophil complexes are important for subsequent neutrophil activation and migration. J Appl Physiol 109: 758–767. Intravital Visualization of Platelets 33. Langer HF, Daub K, Braun G, Schonberger T, May AE, et al. (2007) Platelets recruit human dendritic cells via Mac-1/JAM-C interaction and modulate dendritic cell function in vitro. Arterioscler Thromb Vasc Biol 27: 1463–1470. 34. Shenkman B, Brill G, Solpov A, Vitkovsky Y, Kuznik B, et al. (2006) CD4+ lymphocytes require platelets for adhesion to immobilized fibronectin in flow: role of beta(1) (CD29)-, beta(2) (CD18)-related integrins and non-integrin receptors. Cell Immunol 242: 52–59. 35. McDonald B, McAvoy EF, Lam F, Gill V, de la Motte C, et al. (2008) Interaction of CD44 and hyaluronan is the dominant mechanism for neutrophil sequestration in inflamed liver sinusoids. J Exp Med 205: 915–927. 36. Menezes GB, Lee WY, Zhou H, Waterhouse CC, Cara DC, et al. (2009) Selective down-regulation of neutrophil Mac-1 in endotoxemic hepatic microcirculation via IL-10. J Immunol 183: 7557–7568. 37. Pluskota E, Woody NM, Szpak D, Ballantyne CM, Soloviev DA, et al. (2008) Expression, activation, and function of integrin alphaMbeta2 (Mac-1) on neutrophil-derived microparticles. Blood 112: 2327–2335. 38. Fujimi S, Ogura H, Tanaka H, Koh T, Hosotsubo H, et al. (2002) Activated polymorphonuclear leukocytes enhance production of leukocyte microparticles with increased adhesion molecules in patients with sepsis. J Trauma 52: 443–448. 39. Piccin A, Murphy WG, Smith OP (2007) Circulating microparticles: pathophysiology and clinical implications. Blood Rev 21: 157–171. 40. Ding ZM, Babensee JE, Simon SI, Lu H, Perrard JL, et al. (1999) Relative contribution of LFA-1 and Mac-1 to neutrophil adhesion and migration. J Immunol 163: 5029–5038. 41. Kinoshita M, Uchida T, Sato A, Nakashima M, Nakashima, et al. (2010) Characterization of two F4/80-positive Kupffer cell subsets by their function and phenotype in mice. J Hepatol 53: 903–910. 42. Yan J, Vetvicka V, Xia Y, Hanikyrova M, Mayadas TN, et al. (2000) Critical role of Kupffer cell CR3 (CD11b/CD18) in the clearance of IgM-opsonized erythrocytes or soluble beta-glucan. Immunopharmacology 46: 39–54. 43. Myers KJ, Eppihimer MJ, Hall L, Wolitzky B (1998) Interleukin-12-induced adhesion molecule expression in murine liver. Am J Pathol 152: 457–468. 44. Miyake S, Sakurai T, Okumura K, Yagita H (1994) Identification of collagen and laminin receptor integrins on murine T lymphocytes. Eur J Immunol 24: 2000–2005. 45. Gawaz M, Fateh-Moghadam S, Pilz G, Gurland HJ, Werdan K (1995) Platelet activation and interaction with leucocytes in patients with sepsis or multiple organ failure. Eur J Clin Invest 25: 843–851. 46. Gawaz M, Dickfeld T, Bogner C, Fateh-Moghadam S, Neumann FJ (1997) Platelet function in septic multiple organ dysfunction syndrome. Intensive Care Med 23: 379–385. 47. Li N, Soop A, Sollevi A, Hjemdahl P (2000) Multi-cellular activation in vivo by endotoxin in humans–limited protection by adenosine infusion. Thromb Haemost 84: 381–387. 48. Kalsch T, Elmas E, Nguyen XD, Suvajac N, Kluter H, et al. (2007) Endotoxin- induced effects on platelets and monocytes in an in vivo model of inflammation. Basic Res Cardiol 102: 460–466. 49. Lee WY, Moriarty TJ, Wong CH, Zhou H, Strieter RM, et al. (2010) An intravascular immune response to Borrelia burgdorferi involves Kupffer cells and iNKT cells. Nat Immunol 11: 295–302. 50. Van Rooijen N, Sanders A (1994) Liposome mediated depletion of macrophages: mechanism of action, preparation of liposomes and applications. J Immunol Methods 174: 83–93. 51. Wong J, Johnston B, Lee SS, Bullard DC, Smith CW, et al. (1997) A minimal role for selectins in the recruitment of leukocytes into the inflamed liver microvasculature. J Clin Invest 99: 2782–2790. 52. Liu L, Cara DC, Kaur J, Raharjo E, Mullaly SC, et al. (2005) LSP1 is an endothelial gatekeeper of leukocyte transendothelial migration. J Exp Med 201: 409–418. PLoS ONE | www.plosone.org 13 September 2011 | Volume 6 | Issue 9 | e25109