1. Volume 90, Issue 10, Pages 3783-3795 (May 2006)
Infrared Spectroscopy with Multivariate Analysis Potentially Facilitates the Segregation of Different Types of Prostate Cell 
Matthew J. German, Azzedine Hammiche, Narasimhan Ragavan, Mark J. Tobin, Leanne J. Cooper, Shyam S. Matanhelia, Andrew C. Hindley, Caroline M. Nicholson, Nigel J. Fullwood, Hubert M. Pollock, Francis L. Martin 
Biophysical Journal 
Volume 90, Issue 10, Pages (May 2006)
DOI: /biophysj
Copyright © 2006 The Biophysical Society Terms and Conditions

2. Figure 1
Synchrotron FTIR microspectroscopy to interrogate prostate cells (epithelial versus stromal) in situ. Employing electron microscopy and light microscopy, prostatic glandular architecture was noted. Exploiting the phase contrast facility of the Thermo Nicolet continuum microscope, the location of the 10μm×10μm aperture was tracked. Photomicrographs are of (A) electron microscopy of PZ glandular epithelial in cancer-free tissue (PEC4; scale bar=5μm); (B) electron microscopy of glandular epithelial cells in cancerous tissue (PEC4; scale bar=10μm); (C) H&E of glandular epithelial cells (PZ) derived from cancer-free tissue (PEC4); (D) H&E of glandular elements suspended in stroma (PZ) and derived from cancer-free tissue (PEC4); (E) H&E of a TZ glandular element derived from cancer-free tissue (PEC4); (F) phase contrast image of a glandular element (TZ) derived from cancer-free tissue (PEC4); (G) phase contrast image of glandular elements (TZ) derived from cancerous tissue (PEC4); and (H) phase contrast image of stroma (TZ) located in cancer-free tissue (PEC4). Images were obtained by employing conventional methods.
Biophysical Journal  , DOI: ( /biophysj )
Copyright © 2006 The Biophysical Society Terms and Conditions

3. Figure 2
Epithelial cell spectra derived from different tissue regions employing either ATR spectroscopy (A, C, E, G, I, and K) or synchrotron FTIR microspectroscopy (B, D, F, H, J, and L). From paraffin-embedded blocks, 10-μm-thick sections were mounted on suitable windows to facilitate IR spectra acquisition before being dewaxed and air dried. Multiple spectra (each from a different location) were acquired from PZ (in black), TZ (in blue), or CaP (in red) regions of prostate tissue sets from six individuals (PEC1–PEC6, as detailed in each panel). Individual spectra were normalized to amide II (≈1533cm−1).
Biophysical Journal  , DOI: ( /biophysj )
Copyright © 2006 The Biophysical Society Terms and Conditions

4. Figure 3
Stromal spectra derived from different tissue regions employing either ATR spectroscopy (A, C, E, G, I, and K) or synchrotron FTIR microspectroscopy (B, D, F, H, J, and L). From paraffin-embedded blocks, 10-μm-thick sections were mounted on suitable windows to facilitate IR spectra acquisition before being dewaxed and air dried. Multiple spectra (each from a different location) were acquired from PZ (in black), TZ (in blue), or CaP (in red) regions of prostate tissue sets from six individuals (PEC1–PEC6, as detailed in each panel). Individual spectra were normalized to amide II (≈1533cm−1).
Biophysical Journal  , DOI: ( /biophysj )
Copyright © 2006 The Biophysical Society Terms and Conditions

5. Figure 4
Median spectra (A and C) and cluster vectors (B and D) for all spectra acquired either from epithelial cells lining glandular elements or stroma after ATR spectroscopy or synchrotron FTIR microspectroscopy. Spectra were collected and medians were derived from all the spectra acquired for a particular tissue region (PZ, black line; TZ, blue line; and CaP, red line). Cluster vectors examining loadings as a function of wavenumber were plotted as described (see Supplementary Material).
Biophysical Journal  , DOI: ( /biophysj )
Copyright © 2006 The Biophysical Society Terms and Conditions

6. Figure 5
3-D scores plots on PCs selected to demonstrate best segregation of epithelial cell spectra derived from the different tissue regions. Spectra were collected using synchrotron FTIR microspectroscopy. Each spectrum for each tissue region (PZ, black circles; TZ, blue squares; CaP, red triangles) was expressed in terms of chosen PCs using Pirouette software and rotated to identify segregation of different clusters. Each symbol represents a single spectrum as a single point in “hyperspace”. The 3-D scores plots represent the following: (A) PEC1 on PC1, PC2, and PC4; (B) PEC2 on PC1, PC4, and PC5; (C) PEC3 on PC1, PC2, and PC6; (D) PEC4 on PC2, PC3, and PC4; (E) PEC5 on PC2, PC3, and PC5; and (F) PEC6 on PC1, PC4, and PC5.
Biophysical Journal  , DOI: ( /biophysj )
Copyright © 2006 The Biophysical Society Terms and Conditions

7. Figure 6
2-D plots of the ratios for carbohydrate (quantified by the integrated absorbance in the 900–1185cm−1 region) to phosphates of nucleic acids (quantified by the integrated absorbance in the 1185–1300cm−1 region) versus RNA/DNA (based on the intensity at wavenumber 1121cm−1/intensity at 1020cm−1). Each symbol represents the mean±SD of all the spectrally derived estimations for one tissue region (PZ, TZ, or CaP). The 2-D plots represent the following: (A) spectrally derived estimations for PZ; (B) spectrally derived estimations for TZ; and (C) spectrally derived estimations for CaP. These estimations were derived using OPUS software.
Biophysical Journal  , DOI: ( /biophysj )
Copyright © 2006 The Biophysical Society Terms and Conditions

8. Figure 7
Median spectra and 3-D scores plots on PCs selected to demonstrate segregation of spectra derived from epithelial cells of PZ versus TZ isolated from a cystoprostatectomy specimen (PEC7). Spectra were collected using synchrotron FTIR microspectroscopy, and medians were derived from all the spectra acquired for a particular tissue region (PZ, black line; TZ, blue line). Each spectrum for each tissue region (PZ, black circles TZ, blue squares) were expressed in terms of chosen PCs using the Pirouette software and rotated to identify segregation of different clusters.
Biophysical Journal  , DOI: ( /biophysj )
Copyright © 2006 The Biophysical Society Terms and Conditions