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Detection of a single 4T1-luc2-1A4 cell in vivo.

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posted on 2010-02-23, 00:41 authored by Jae-Beom Kim, Konnie Urban, Edward Cochran, Steve Lee, Angel Ang, Bradley Rice, Adam Bata, Kenneth Campbell, Richard Coffee, Alex Gorodinsky, Zhan Lu, He Zhou, Takashi Kei Kishimoto, Peter Lassota

(A-C) Bioluminescent signal of a single 4T1-luc2-1A4 cell in vivo. A single cell was implanted into the back of a nu/nu mouse. D-luciferin was injected into the mouse intraperitoneally and bioluminescent images were taken using an IVIS Spectrum (FOV; C, binning; small, f stop; 1, exposure time; 5 min). Images for pre- and post-luciferin injection were shown in panels (A) and (B), respectively. Magnified image of the dotted area from panel (B) is shown on panel (C). The dotted circle represents the single cell signal. The asterisk (*) indicates the background signal from the gut. (D,E) Line profiling analysis of single cell signal. Light emission was plotted along the line shown on panel (D). Peak signal in the panel (E) represents the light emission from a single 4T1-luc2-1A4 cell. (F) Tumor development from five 4T1-luc2-1A4 cells. Cells were implanted subcutaneously (using a micropipette) into the dorsal flank region of a nu/nu mouse. Bioluminescent images were first taken before the D-luciferin injection (Pre-luciferin). The animal was then imaged on day 0 through day 42. (G) Monitoring of tumor growth from 5 cells of 4T1-luc2-1A4. Bioluminescent signals were quantified using Living Image software and plotted against physical tumor volume measurements by a caliper. Tumor was not palpable till day 27 post-implantation while bioluminescent signals were detected from the day 0. Note that total flux was plotted in a logarithmic scale. (H) Tumor development from 10 cells of 4T1-luc2-1A4. Cells were implanted subcutaneously (using a micropipette) into the back of a nu/nu mouse. The background signal is shown in pre-D-luciferin injection image (Pre-luciferin). The tumor growth was monitored for 40 days using an IVIS Spectrum and a caliper. (I) Monitoring of tumor growth from 10 cells of 4T1-luc2-1A4. Tumor volumes were measured using a caliper and plotted against bioluminescent signals which were quantified using Living Image software. Tumor was not palpable till day 29 after implantation. On the contrary, bioluminescent signals were distinct from the day 0 of implantation.

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