Waiting
Login processing...

Trial ends in Request Full Access Tell Your Colleague About Jove
Click here for the English version

Immunology and Infection

Intravital Imaging av Mouse Thymus med 2-Photon Mikroskopi

Published: January 7, 2012 doi: 10.3791/3504

Materials

Name Type Company Catalog Number Comments
Rhodamine B ishothiocyanate-Dextran Sigma-Aldrich R9379 prepare stock at 20 mg/ml
Two-photon microscope Prairie Technologies Inc. Prairie Ultima X-Y
Ti:Sapphire laser Coherent, Inc. Chameleon Ultra Family
20x/1.00 NA immersion objective Olympus Inc. XLUMPLFLN 20XW
Holder (Filters/Dichroic) Chroma Technology Corp. 91018 BX2 (U-MF2)
525 nm/50 filter Chroma Technology Corp. ET525/50m
595 nm/50 filter Chroma Technology Corp. ET595/50m
565 nm dichroic Chroma Technology Corp. 565dcxr
Imaris software Bitplane AG Inc. Imaris
Volocity PerkinElmer Inc. Volocity
ImageJ NIH, USA ImageJ

DOWNLOAD MATERIALS LIST

References

  1. Bettelli, E. Reciprocal developmental pathways for the generation of pathogenic effector TH17 and regulatory T cells. Nature. 441, 235-238 (2006).
  2. Ramsdell, F., Zúñiga-Pflücker, J. C., Takahama, Y. In vitro systems for the study of T cell development: fetal thymus organ culture and OP9-DL1 cell coculture. Current protocols in immunology. Coligan, J. E. , (2006).
  3. Tadokoro, C. E. Regulatory T cells inhibit stable contacts between CD4+ T cells and dendritic cells in vivo. J. Exp. Med. 203, 505-511 (2006).
  4. 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, 154-159 (2004).
  5. Miller, M. J. Imaging the single cell dynamics of CD4+ T cell activation by dendritic cells in lymph nodes. J. Exp. Med. 200, 847-856 (2004).
  6. Shakhar, G. Stable T cell-dendritic cell interactions precede the development of both tolerance and immunity in vivo. Nat. Immunol. 6, 707-714 (2005).
  7. Hugues, S. Distinct T cell dynamics in lymph nodes during the induction of tolerance and immunity. 5, 1235-1242 (2004).
  8. Tang, Q. Visualizing regulatory T cell control of autoimmune responses in nonobese diabetic mice. Nat. Immunol. 7, 83-92 (2006).
  9. Le Borgne, M. The impact of negative selection on thymocyte migration in the medulla. Nature immunology. 10, 823-830 (2009).
  10. Baez, S. An open cremaster muscle preparation for the study of blood vessels by in vivo microscopy. Microvascular research. 5, 384-394 (1973).
  11. Wang, B., Zinselmeyer, B. H., McDole, J. R., Gieselman, P. A., Miller, M. J. Non-invasive Imaging of Leukocyte Homing and Migration in vivo. J. Vis. Exp. (46), e2062-e2062 (2010).
  12. Barretto, R. P. J. Time-lapse imaging of disease progression in deep brain areas using fluorescence microendoscopy. Nature medicine. 17, 223-228 (2011).
Intravital Imaging av Mouse Thymus med 2-Photon Mikroskopi
Play Video
PDF DOI DOWNLOAD MATERIALS LIST

Cite this Article

Caetano, S. S., Teixeira, T.,More

Caetano, S. S., Teixeira, T., Tadokoro, C. E. Intravital Imaging of the Mouse Thymus using 2-Photon Microscopy. J. Vis. Exp. (59), e3504, doi:10.3791/3504 (2012).

Less
Copy Citation Download Citation Reprints and Permissions
View Video

Get cutting-edge science videos from JoVE sent straight to your inbox every month.

Waiting X
Simple Hit Counter