Correlation based on Sequential LM and EM imaging to Target a Single Tumor Cell.

Abstract

<p>A. 3D view of a 2PEM z-stack of mouse ear skin tissue, 7 days post-injection with GFP-expressing tumor cells (green). Vessels are stained with Evans Blue (red). The arrowhead indicates the part of the blood vessel that is also visible in the thick section shown in panel D. The image is obtained using the 3D Viewer plugin in Fiji. Scale bar: 50 µm. See <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0114448#pone.0114448.s004" target="_blank">Movie S2</a>. B. 3D model of the imaged volume. The cell of interest, boxed in panel A and B, is segmented in green, the other cells are segmented in grey. The vessels are shown in red. Scale bar: 50 µm. C. Cartoon of the sectioning procedure. The resin embedded sample is sectioned from the direction indicated with the arrow. To approach the ROI, 180 thick sections (500 nm) were obtained from the sample ('Approach and Correlation'). Next, 10 consecutive series of 10 thin (60 nm) and two thick (500 nm) sections were obtained from the ROI. Thick sections were used for correlation and the thin sections were imaged with EM. D. The cell of interest (box) was identified in a thick toluidine blue stained section of the ROI. The arrowhead points to a cross-section of the vessel depicted in A with a white arrowhead. Scale bar: 50 µm. E: Low magnification EM image of the cell boxed in panel D. The cell appears highly polarized and contains protrusive structures, showed at higher magnification in panel F. Scale bar in E: 5 µm, in F: 2 µm. See <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0114448#pone.0114448.s004" target="_blank">Movie S2</a>.</p