© 2007 Nature Publishing Group

Supplemental figures for

All-optical anatomical coregistration for small animal molecular imaging using dynamic contrast.

Elizabeth M. C. Hillman

1 and Anna Moore

2

1Laboratory for Functional Optical Imaging, Department of Biomedical Engineering, Columbia University, 1210

Amsterdam Avenue, New York, NY 10027. 1-212-854-2788, eh2245@columbia.edu.

2Molecular Imaging Laboratory, MGH/MIT/HMS Athinoula A. Martinos Center for Biomedical Imaging, Massachusetts

General Hospital, Building 149, 13th

Street, Charlestown, MA 02129

Figure S1. Raw timecourse images of indocyanine green and dextran texas red following simultaneous injection

Figure S2. In-vivo anatomical maps derived using PCA of a 20 second image series following ICG injection

Figure S3. In-vivo optical spatiotemporal separation of nine ‘organs’ using basis timecourses (ICG only).

Figure S4. Comparison of DFMI anatomical image with digital anatomical atlas.

© 2007 Nature Publishing Group

Figure S1. Raw timecourse images of indocyanine green and dextran texas red following

simultaneous injection. Left: Raw data series of ICG and Texas red from injection to 2 minutes post-

injection. Mouse is prone. Each image is normalized to its own maximum. Right: Timecourses of

selected regions are shown for nominally identified organs. Top row shows absolute timecourse,

bottom row shows time-course divided by the mean time-course. Note high initial fluorescence in large

intestine at texas-red wavelengths (corresponding to food autofluorescence). Note the rapid transit of

ICG in the lungs (as the direct route from the tail vein to the right ventricle to the pulmonary artery).

© 2007 Nature Publishing Group

Figure S2. In-vivo anatomical maps derived using PCA of an image series following ICG

injection (same as figure 2 in main text, but for first 20 seconds of mouse A). Left: maps of the 2nd

, 3rd

and 4th

temporal principal components of images acquired during the first 20 seconds after ICG

injection (color-coded red, green and blue). The mouse is in a prone position. Right: The

corresponding PCA basis timecourses of the 1st – 4

th components of the image series. Each identified

organ has a different red-green-blue combination, and hence each has its own distinctive timecourse.

The spikes in the 4th component correspond to breathing-related motion. Labeled organs were

identified by comparison with post-mortem dissection and general anatomy.

© 2007 Nature Publishing Group

Figure S3. In-vivo optical spatiotemporal separation of nine ‘organs’ using basis timecourses

(ICG only). The individual spatial maps resulting from a non-negative least-squares fit of 9 basis

timecourses to a 26 minute long image series (these are the individual elements of the image

composition shown in figure 3 of the main text). Fit results are shaded cyan and overlaid onto a feint

bright-field image. In most cases, localization to one place is achieved. Additional structure indicates

that these regions also have similar dynamic behavior to the primary region.

© 2007 Nature Publishing Group

Figure S4. Comparison of DFMI anatomical image with digital anatomical atlas. Top: image from

figure 3 in main test. Bottom: Digimouse anatomical atlas http://neuroimage.usc.edu/Digimouse.html

red = liver, blue = spleen, purple = kidney, green = heart, pink = brain, cyan = lungs (pale blue =

bladder, yellow = testes [digimouse is male, dynamouse is female]).