Tracking Neutrophil Intraluminal Crawling, Transendothelial Migration and Chemotaxis in Tissue by Intravital Video Microscopy

JoVE 3296 9/24/2011

Najia Xu, Xi Lei, Lixin Liu

Department of Pharmacology, University of Saskatchewan

We describe a protocol of brightfield intravital microscopy for measuring dynamic neutrophil-endothelial cell interactions during neutrophil recruitment in response to the source of a neutrophil chemoattractant in vivo. Neutrophil intraluminal crawling, transendothelial migration and chemotaxis in mouse cremaster muscle tissue are visualized with time-lapsed video photography and tracked with ImageJ.

The Mouse Cremaster Muscle Preparation for Intravital Imaging of the Microcirculation

JoVE 2874 6/10/2011

Pooneh Bagher¹, Steven S. Segal^1,2

¹Department of Medical Pharmacology and Physiology, University of Missouri, ²Dalton Cardiovascular Research Center, University of Missouri

A tissue preparation is described for visualization and experimental manipulation of the living microcirculation. In anesthetized male mice, the thin, highly vascularized cremaster muscle is prepared for intravital microscopy to study microvascular networks including arterioles, capillaries and venules. This preparation is readily adapted for rats and hamsters.

Evaluation of Nanoparticle Uptake in Tumors in Real Time Using Intravital Imaging

JoVE 2808 6/21/2011

Choi-Fong Cho¹, Amber Ablack², Hon-Sing Leong², Andries Zijlstra³, John Lewis^2,4

¹Department of Medical Biophysics, University of Western Ontario, ²London Regional Cancer Program, London Health Science Centre, ³Department of Pathology, Vanderbilt University , ⁴Translational Prostate Cancer Research Group, London Health Science Centre

We present a novel approach to quantify nanoparticle localization in the vasculature of human xenografted tumors using dynamic, real-time intravital imaging in an avian embryo model.

Intravital Microscopy of the Mouse Brain Microcirculation using a Closed Cranial Window

JoVE 2184 11/18/2010

Pedro Cabrales¹, Leonardo J. M. Carvalho²

¹Bioengineering, University of California, San Diego, ²La Jolla Bioengineering Institute

Intravital microscopy to follow temporal and spatial hemodynamic and inflammatory events in the pial microcirculation.

Intravital Microscopy of the Inguinal Lymph Node

JoVE 2551 4/04/2011

Stephanie L. Sellers¹, Geoffrey W. Payne²

¹Interdisciplinary Science, University of Northern British Columbia, ²Northern Medical Program, University of Northern British Columbia

A technique for performing intravital microscopy of the inguinal lymph node (LN) is outlined. Such technique allows for real-time, in vivo study of the lymph node microvasculature and structure both during homeostasis and infection. This technique can be adapted to cell trafficking studies and to other lymph node sites.

Intravital Microscopy of the Spleen: Quantitative Analysis of Parasite Mobility and Blood Flow

JoVE 3609 1/14/2012

Mireia Ferrer*¹, Lorena Martin-Jaular*¹, Maria Calvo², Hernando A. del Portillo^1,3

¹Department of poverty related diseases, Barcelona Centre for International Health Research, ²Confocal Microscopy Unit, University of Barcelona- Scientific and Technological Centers, ³Institució Catalana de Recerca i Estudis Avançats (ICREA)

We show the method for performing intravital microscopy of the spleen using GFP transgenic malaria parasites and the quantification of parasite mobility and blood flow within this organ.

Intravital Imaging of the Mouse Thymus using 2-Photon Microscopy

JoVE 3504 1/07/2012

Susana S. Caetano, Tatiana Teixeira, Carlos E. Tadokoro

Laboratory of Immune Regulation, Instituto Gulbenkian de Ciência

We have developed novel laboratory tools and protocols for intravital imaging acquisition of the thymus. Our technique should help in the identification of “niches” within the thymus where T cell development occurs.

June 2011: This Month in JoVE

JoVE 3549 6/01/2011

Aaron Kolski-Andreaco

Intravital Imaging of the Mouse Popliteal Lymph Node

JoVE 3720 2/08/2012

H. L. Rachel Liou¹, Jay T. Myers¹, Deborah S. Barkauskas¹, Alex Y. Huang²

¹Department of Pediatrics, Case Western Reserve University , ²Department of Pediatrics, Pathology and Biomedical Engineering, Case Western Reserve University

Recent advances in 2-photon microscopy have enabled real-time in situ imaging of live tissues in animal models, thereby enhancing our ability to investigate cellular behavior in both physiologic and pathologic conditions. Here, we outline the preparations required to perform intravital imaging of the mouse popliteal lymph node.

Dendra2 Photoswitching through the Mammary Imaging Window

JoVE 1278 6/05/2009

Bojana Gligorijevic ^1,2, Dmitriy Kedrin¹, Jeffrey E Segall¹, John Condeelis^1,2, Jacco van Rheenen^1,2,3

¹Department of Anatomy and Structural Biology, Albert Einstein College of Medicine - Yeshiva University, ²Gruss Lipper Biophotonics Center, Albert Einstein College of Medicine - Yeshiva University, ³Hubrecht Institute-KNAW and University Medical Center Utrecht

Intravital photoswitching and tracking of Dendra2-labeled tumor cells through the Mammary Imaging Window is a technique which allows us to image the metastatic behavior of tumor cells in chosen tumor microenvironments over a timescale of days.