Movies

Imaging of CMTMR labeled breast carcinoma cells (red) and their surrounding neutrophils by light sheet microscopy of whole lungs

Endogenous neutrophils were labeled 5 mins before harvesting the lungs with Alexa 647-labeled anti-Ly6G mAb. Lungs were inflated with low melting temperature agarose, excised and cleared. Autofluorescent lung stromal cells are depicted in green. Bar= 100µm.

Stress fiber bending by the nucleus of a transmigrating effector T cell crossing transcellularly through inflamed HDMVECs

A time lapse movie depicting a Hoechst labeled effector T cell crossing through a transcellular route of IL-1b-stimulated HDMVECs expressing RFP-Lifeact. Images were taken 20 sec apart. Elapsed time is designated as h:mm:ss. Bar, 5 μm.

Migration of Hoechst labeled granulocyte-like cells across inflamed endothelial cells

Blue: nucleus. The red line depicts the deformed nucleus and the green line depicts the edge of the leading edge of the transmigrating granulocyte extended underneath the endothelial cell.

Hoechst labeled breast cancer cell (E0771) in the process of crossing a confluent monolayer of bEnd3 murine endothelial cells.

Phase contrast and fluorescence microscopy. Note the very slow generation of a sub-endothelial leading edge by the transmigrating tumor cell followed by a slow squeezing of the tumor nucleus through the endothelial junction.

Adoptively transferred melanoma cells inside and outside the lung vasculature

3D image of lungs imaged by light sheet microscopy (LSM). Recipient mice were injected with 20,000 B16-F10 labeled with CMTMR 1 hour before (red). Green: auto fluorescence of the lung stroma.

Melanoma cell in the process of extravasation through a lung vessel

3D image of lungs imaged by light sheet microscopy (LSM). Recipient mice were injected with 20,000 B16-F10 labeled with CMTMR 3 hours before. To visualize lung vessels, Alexa 647 conjugated anti CD31 mAb was injected 5 minutes before lungs were harvested, fixed and cleared for LSM imaging.

Naïve CD4 T cells arrest on antigen presenting DCs in vivo independently of DC ICAM-1

A dsRed labeled OT-II CD4+ cell (red) interacting with an ICAM-1 and 2 double deficient DC (GFP, green) in the T zone of a popliteal lymph node. WT DCs are CFP labeled. Mice were immunized with αDEC-205-OVA and αCD40 mAb. Bar, 10 μm.

Infection of mCherry encoding influenza virus in the lung airways

3D image of lungs infected with the virus imaged by light sheet microscopy (LSM) 4 days post infection. The influenza infected airways are depicted in red. Green: auto fluorescence of the lung stroma. Note the high specificity of the virus towards the airways and the large peribronchial vessel (green) surrounding the virus infected bronchi (red).