Jeffrey Segall's Lab

Kedrin et al., 2008 Nature Methods

Intravital imaging of metastatic behavior through a mammary imaging window

Nat Methods. 2008 Dec;5(12):1019-21.

Kedrin D, Gligorijevic B, Wyckoff J, Verkhusha VV, Condeelis J, Segall JE, van Rheenen J.

Department of Anatomy and Structural Biology, Albert Einstein College of Medicine of Yeshiva University

We report a technique to evaluate the same tumor microenvironment over multiple intravital imaging sessions in living mice. We optically marked individual tumor cells expressing photoswitchable proteins in an orthotopic mammary carcinoma and followed them for extended periods through a mammary imaging window. We found that two distinct microenvironments in the same orthotopic mammary tumor affected differently the invasion and intravasation of tumor cells.

Supplementary Movie 1

Movie recorded through the ocular, at low magnification. After focusing the field and checking for the flowing vessels in the green epifluorescence channel, the photoswitched area can be recognized in the red channel.

Supplementary Movie 2

120-minute timelapse of a 250 um 250um area photoswitched inside the tumor. Notice the cells which move towards the fast flowing vessel. Non-photoswitched cells (green), photoswitched cells (red) and matrix (blue).

 

 

Supplementary Movie 3

Z-stack of collagen (blue) and Dendra2-MTLn3 tumor cells (green and red) visualized in the live animal using a multiphoton microscope. The stepsize is 5um.

 

Supplementary Movie 4

10-minute timelapse of flowing vessels labeled using AlexaFluo647-10K (red, moving shadows are red blood cells) and Dendra2-MTLn3 tumor cells (green). Other channels were omitted for better contrast.