DOI: 10.15129/ef11d3c5-7c59-49b9-8009-143e52daa017
title: Data for: "Through Tissue Imaging of a Live Breast Cancer 3D Tumor Model Using Handheld Surface Enhanced Spatially Offset Resonance Raman Spectroscopy"

description: Through Tissue Imaging of a Live Breast Cancer 3D Tumor Model Using Handheld Surface Enhanced Spatially Offset Resonance Raman Spectroscopy

The purpose of this data set was to image a 3D tumor model through 15 mm of tissue using handheld surface enhanced spatially offset resonance Raman spectroscopy. By using this technique it is possible to image at depths using Raman spectroscopy. 

Abbreviations 
SORS = spatially offset Raman spectroscopy 
SERS = Surface enhanced Raman spectroscopy
SERRS = Surface enhanced resonance Raman spectroscopy
SESORS = Surface enhanced spatially offset Raman spectroscopy
SESORRS = Surface enhanced spatially offset resonance Raman spectroscopy
Gold nanoparticles = AuNPs
MTS = multicellular tumor spheroids

All data can be found in the database file “handheldDB”. The database can be opened using custom software from cobalt light systems. The customized software was purpose wrote and confidential however all data used is shown in the excel spreadsheets with respect to the corresponding figures. 


Figure 1 Figure 1- (a – e) Chemical structure of dye676, dye823, dye959, BPE and AZPY respectively. Chemical structures were drawn using ChemDraw (f) Bar chart showing average peak intensities of dye676, dye823, dye959, BPE and AZPY at 1598, 1592, 1572, 1201 and 1162 cm-1 respectively, as well as the relative percentage peak intensity, through 5 mm of tissue. Nanotag solutions were held in a cuvette and the cuvette was placed behind tissue samples. Spectra were collected using a handheld SORS instrument with 830 nm laser excitation at an 8 mm offset. Peak intensities were obtained by scanning 3 replicate samples, 5 times (3 second acquisitions, 5 accumulations). The average peak intensity for each of the 5 dyes is shown and error bars represent ± one standard deviation. Measurements were performed using a cobalt light systems resolve instrument 830 nm. Spectra were produced from a database file using specialized software from Cobalt light systems. The resulting spectra can be seen in the excel spreadsheets. The customized software was purpose wrote and confidential.

Figure 2 (a) A false color 2D heat SESORRS map of MTS containing dye823 through 15 mm of tissue. The map was constructed using the peak intensity at 1178 cm-1, measurements were carried out using 3 mm step to create an image of 7 x 7 pixels. Spectra were truncated, baselined and smoothed prior to processing. A combination surface/contour false color was used to generate a 2D heat map and show the tracking of the MTS through 15 mm of tissue. The 2D false color map was created in Matlab using customized scripts. Clear discrimination is seen between spectra collected at the point of maximum intensity where the nanotags were spotted and that collected where the nanotags were not present. (b) The corresponding maximum and minimum collected 8 mm offset spectra.  All measurements were carried out using a 3 s integration time, 5 accumulations, 830 nm laser excitation wavelength. Measurements were performed using a cobalt light systems resolve instrument 830 nm. Spectra were produced from a database file using specialized software from Cobalt light systems. The resulting spectra can be seen in the excel spreadsheets. The customized software was purpose wrote and confidential.

Figure 3 - The tracking of dye 823 nanotag solution through 25 mm of tissue. The tissue and dye823 reference spectra are shown at the bottom and top respectively. The middle spectrum represents the Raman signal collected at an 8 mm offset through 25 mm of tissue. The peak at 1178 cm-1 is easily detectable by eye and the peak at 1592 cm-1 is also detectable, albeit to a lesser extent. All measurements were carried out using a 3 s integration time, 5 accumulations, 830 nm laser excitation wavelength. Measurements were performed using a cobalt light systems resolve instrument 830 nm. Spectra were produced from a database file using specialized software from Cobalt light systems. The resulting spectra can be seen in the excel spreadsheets. The customized software was purpose wrote and confidential.

Figure S1 – SEM Image of gold nanoparticles. Scanning electron microscope (SEM) images were obtained using a FEI Sirion 200 ultra-high resolution Schottky field emission scanning electron microscope with FEI software. 

Figure S2 - SERS spectra of the five Raman reporters used in this work. Dyes 676, 823 and 959 are chalcogen based reporters BPE and AZPY are commercially available. Spectra were obtained using the SORS instrument in a conventional Raman mode. Dyes 1 – 3 are resonant at 676, 823 and 959 nm respectively. BPE and AZPY are non-resonant molecules. Spectra of each nanotag solution (3 samples, 5 replicates, per Raman reporter) were collected. All measurements were carried out using a 3 s integration time, 5 accumulations, 830 nm laser excitation wavelength. Spectra were baselined and normalized using custom scripts in MATLAB. 


Figure S3 – Experimental set up

Figure S4 – experimental set up

Figure S5 –The tracking of dye 823 nanotag solution through 20-25 mm of tissue following a scaled subtraction. The peaks at 1178 cm-1 and 1592 cm-1 are revealed. This approach removes signal collected at the zero position (tissue) from that collected at the 8 mm offset (tissue and dye). All measurements were carried out using a 3 s integration time, 5 accumulations, 830 nm laser excitation wavelength. Spectra of dye823 obscured by 25 mm of pork tissue (3 samples, 5 replicates) were collected to produce two data sets. One set of spectra (total 15) were collected at the zero position, the other set at the 8 mm offset position.  These two data sets were used to carry out a scaled subtraction using custom scripts in written for MATLAB.