Abstract
Intravital multiphoton microscopy (MP-IVM) is a powerful imaging approach that allows direct visualization of cells within their native environment in real time. Multiphoton imaging of immune cells has been performed in different tissues, and these studies have revealed intriguing insights into the unique migratory and interactive behavior of immune cells in the steady-state and during disease conditions. Here we provide an overview of a MP-IVM model of the mouse ear skin, as well as the benefits, limitations and pitfalls of this approach. We also discuss the prospects of intravital imaging in the areas of image analysis, data management and standardization.
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References
Banchereau, J., Steinman, R.M.: Dendritic cells and the control of immunity. Nature 392(6673), 245–252 (1998), doi:10.1038/32588
Bonasio, R., von Andrian, U.H.: Generation, migration and function of circulating dendritic cells. Curr. Opin. Immunol. 18(4), 503–511 (2006); doi:S0952-7915(06)00106-3 [pii] 10.1016/j.coi.2006.05.011
Bousso, P., Bhakta, N.R., Lewis, R.S., Robey, E.: Dynamics of thymocyte-stromal cell interactions visualized by two-photon microscopy. Science 296(5574), 1876–1880 (2002); doi:10.1126/science.1070945 296/5574/1876 [pii]
Cahalan, M.D., Parker, I.: Choreography of cell motility and interaction dynamics imaged by two-photon microscopy in lymphoid organs. Annu. Rev. Immunol. 26, 585–626 (2008); doi:10.1146/annurev.immunol.24.021605.090620
Dickinson, M.E., Bearman, G., Tille, S., Lansford, R., Fraser, S.E.: Multi-spectral imaging and linear unmixing add a whole new dimension to laser scanning fluorescence microscopy. Biotechniques 31(6),1272, 1274–1276, 1278 (2001)
Ducros, M., Moreaux, L., Bradley, J., Tiret, P., Griesbeck, O., Charpak, S.: Spectral unmixing: analysis of performance in the olfactory bulb in vivo. PLoS One 4(2), e4418 (2009); doi:10.1371/journal.pone.0004418
Faust, N., Varas, F., Kelly, L.M., Heck, S., Graf, T.: Insertion of enhanced green fluorescent protein into the lysozyme gene creates mice with green fluorescent granulocytes and macrophages. Blood 96(2), 719–726 (2000)
Gebhardt, T., Wakim, L.M., Eidsmo, L., Reading, P.C., Heath, W.R., Carbone, F.R.: Memory T cells in nonlymphoid tissue that provide enhanced local immunity during infection with herpes simplex virus. Nat. Immunol. 10(5), 524–530 (2009); doi:ni.1718 [pii] 10.1038/ni.1718
Germain, R.N., Miller, M.J., Dustin, M.L., Nussenzweig, M.C.: Dynamic imaging of the immune system: progress, pitfalls and promise. Nat. Rev. Immunol. 6(7), 497–507 (2006); doi:nri1884 [pii] 10.1038/nri1884
Havran, W.L., Allison, J.P.: Developmentally ordered appearance of thymocytes expressing different T-cell antigen receptors. Nature 335(6189), 443–445 (1988); doi:10.1038/335443a0
Hiraoka, Y., Swedlow, J.R., Paddy, M.R., Agard, D.A., Sedat, J.W.: Three-dimensional multiple-wavelength fluorescence microscopy for the structural analysis of biological phenomena. Semin. Cell. Biol. 2(3), 153–165 (1991)
Jung, S., Aliberti, J., Graemmel, P., Sunshine, M.J., Kreutzberg, G.W., Sher, A., Littman, D.R.: Analysis of fractalkine receptor CX(3)CR1 function by targeted deletion and green fluorescent protein reporter gene insertion. Mol. Cell. Biol. 20(11), 4106–4114 (2000)
Kissenpfennig, A., Henri, S., Dubois, B., Laplace-Builhe, C., Perrin, P., Romani, N., Tripp, C.H., Douillard, P., Leserman, L., Kaiserlian, D., Saeland, S., Davoust, J., Malissen, B.: Dynamics and function of Langerhans cells in vivo: dermal dendritic cells colonize lymph node areas distinct from slower migrating Langerhans cells. Immunity 22(5), 643–654 (2005); doi:S1074-7613(05)00131-7 [pii] 10.1016/j.immuni.2005.04.004
Lansford, R., Bearman, G., Fraser, S.E.: Resolution of multiple green fluorescent protein color variants and dyes using two-photon microscopy and imaging spectroscopy. J. Biomed. Opt. 6(3), 311–318 (2001); doi:10.1117/1.1383780
Lindquist, R.L., Shakhar, G., Dudziak, D., Wardemann, H., Eisenreich, T., Dustin, M.L., Nussenzweig, M.C.: Visualizing dendritic cell networks in vivo. Nat. Immunol. 5(12), 1243–1250 (2004); doi:ni1139 [pii] 10.1038/ni1139
Matheu, M.P., Beeton, C., Garcia, A., Chi, V., Rangaraju, S., Safrina, O., Monaghan, K., Uemura, M.I., Li, D., Pal, S., de la Maza, L.M., Monuki, E., Flugel, A., Pennington, M.W., Parker, I., Chandy, K.G., Cahalan, M.D.: Imaging of effector memory T cells during a delayed-type hypersensitivity reaction and suppression by Kv1.3 channel block. Immunity 29(4), 602–614 (2008); doi:S1074-7613(08)00408-1 [pii] 10.1016/j.immuni.2008.07.015
Mempel, T.R., Henrickson, S.E., Von Andrian, U.H.: T-cell priming by dendritic cells in lymph nodes occurs in three distinct phases. Nature 427(6970), 154–159 (2004); doi:10.1038/nature02238
Miller, M.J., Wei, S.H., Parker, I., Cahalan, M.D.: Two-photon imaging of lymphocyte motility and antigen response in intact lymph node. Science 296(5574), 1869–1873 (2002); doi:10.1126/science.1070051
Mohrs, M., Shinkai, K., Mohrs, K., Locksley, R.M.: Analysis of type 2 immunity in vivo with a bicistronic IL-4 reporter. Immunity 15(2), 303–311 (2001); doi:S1074-7613(01)00186-8 [pii]
Nathan, C.: Neutrophils and immunity: challenges and opportunities. Nat. Rev. Immunol. 6(3), 173–182 (2006); doi:nri1785 [pii] 10.1038/nri1785
Ng, L.G., Hsu, A., Mandell, M.A., Roediger, B., Hoeller, C., Mrass, P., Iparraguirre, A., Cavanagh, L.L., Triccas, J.A., Beverley, S.M., Scott, P., Weninger, W.: Migratory dermal dendritic cells act as rapid sensors of protozoan parasites. PLoS Pathog. 4(11), 1000222 (2008), doi:10.1371/journal.ppat.1000222
Peters, N.C., Egen, J.G., Secundino, N., Debrabant, A., Kimblin, N., Kamhawi, S., Lawyer, P., Fay, M.P., Germain, R.N., Sacks, D.: In vivo imaging reveals an essential role for neutrophils in leishmaniasis transmitted by sand flies. Science 321(5891), 970–974 (2008); doi:321/5891/970 [pii] 10.1126/science.1159194
Roediger, B., Ng, L.G., Smith, A.L., de St Groth, B.F., Weninger, W.: Visualizing dendritic cell migration within the skin. Histochem. Cell Biol. 130(6), 1131–1146 (2008); doi:10.1007/s00418-008-0531-7
Sen, D., Forrest, L., Kepler, T.B., Parker, I., Cahalan, M.D.: Selective and site-specific mobilization of dermal dendritic cells and Langerhans cells by Th1- and Th2-polarizing adjuvants. Proc. Natl. Acad. Sci. USA 107(18), 8334–8339 (2010); doi:0912817107 [pii] 10.1073/pnas.0912817107
Sumaria, N., Roediger, B., Ng, L.G., Qin, J., Pinto, R., Cavanagh, L.L., Shklovskaya, E., Fazekas de St Groth, B., Triccas, J.A., Weninger, W.: Cutaneous immunosurveillance by self-renewing dermal gammadelta T cells. J. Exp. Med. 208(3), 505–518 (2011); doi:jem.20101824 [pii] 10.1084/jem.20101824
Swedlow, J.R., Goldberg, I., Brauner, E., Sorger, P.K.: Informatics and quantitative analysis in biological imaging. Science 300(5616), 100–102 (2003); doi:10.1126/science.1082602
Wan, Y.Y., Flavell, R.A.: Identifying Foxp3-expressing suppressor T cells with a bicistronic reporter. Proc. Natl. Acad. Sci. USA 102(14), 5126–5131 (2005); doi:0501701102 [pii] 10.1073/pnas.0501701102
Witte, S., Negrean, A., Lodder, J.C., de Kock, C.P., Testa Silva, G., Mansvelder, H.D., Louise Groot, M.: Label-free live brain imaging and targeted patching with third-harmonic generation microscopy. Proc. Natl. Acad. Sci. USA (2011); doi:1018743108 [pii] 10.1073/pnas.1018743108
Zhang, J., Varas, F., Stadtfeld, M., Heck, S., Faust, N., Graf, T.: CD41-YFP mice allow in vivo labeling of megakaryocytic cells and reveal a subset of platelets hyperreactive to thrombin stimulation. Exp. Hematol. 35(3), 490–499 (2007); doi:S0301-472X(06)00714-4 [pii] 10.1016/j.exphem.2006.11.011
Zinselmeyer, B.H., Lynch, J.N., Zhang, X., Aoshi, T., Miller, M.J.: Video-rate two-photon imaging of mouse footpad - a promising model for studying leukocyte recruitment dynamics during inflammation. Inflamm. Res. 57(3), 93–96 (2008); doi:10.1007/s00011-007-7195-y
Zoumi, A., Yeh, A., Tromberg, B.J.: Imaging cells and extracellular matrix in vivo by using second-harmonic generation and two-photon excited fluorescence. Proc. Natl. Acad. Sci. USA 99(17), 11014–11019 (2002); doi:10.1073/pnas.172368799
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Keeble, J., Goh, C.C., Wang, Y., Weninger, W., Ng, L.G. (2012). Intravital Multiphoton Imaging of Immune Cells. In: Loménie, N., Racoceanu, D., Gouaillard, A. (eds) Advances in Bio-Imaging: From Physics to Signal Understanding Issues. Advances in Intelligent and Soft Computing, vol 120. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-25547-2_1
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