Page 118 - Vibrational Spectroscopic Imaging for Biomedical Applications

94 Cha pte r T h ree

responses of the cell may be a constant between conditions, a hypoth-
esis that requires further testing.
The effects of basic growth media (RPMI and Ham’s F-12) on two
prostate cancer epithelial cell lines does not affect its phenotypic clas-
sification using FTIR. Whether this is the case for other cell types
grown in these media requires further study. However, commonly
used substrates for spectroscopy do influence the cells at the whole
cell level as well as at the molecular level. It was concluded that gelatin-
coated quartz and MirrIR slides may provide the best approach for
long-term cell viability. On thicker/highly concentrated protein-based
biological supports, it was found that optical artifacts can manifest and
these were supported by time-lapse observations.
There have been encouraging results reported within the context
of live cell experiments using FTIR. The collective demonstration of
biochemical stability for different cell types (T-1, HeLa and fibro-
blasts) by the various research groups working within this field, 57–59
57
together with Moss et al. suggestion that correction for water absor-
bance may not be necessary, suggests that future FTIR studies may be
able to measure early biochemical responses of single living cells to
stimuli. Raman-based live cell studies have shown excellent pros-
pects for cell phenotyping as well as probing the distributions of
native biomolecules of a cell with high sensitivity and spatial resolu-
tion and without the requirement for exogenous labeling.

Acknowledgments
Support was received from the Association for International Cancer
Research (AICR Grant number 04-518) and The Prostate Cancer
Foundation during the writing of this article and some of the experi-
ments described within it. We gratefully thank Dr. Stephen Murray
(Paterson Institute for Cancer Research, UK) for use of the time-lapse
video microscope.

References
1. L. Chiriboga, P. Xie, H. Yee, V. Vigorita, D. Zarou, D. Zakim, and M. Diem,
“Infrared Spectroscopy of Human Tissue. I. Differentiation and Maturation of
Epithelial Cells in the Human Cervix,” Biospectroscopy, 4:47–53, 1998.
2. H. Y. N. Holman, M. C. Martin, E. A. Blakely, K. Bjornstad, and W. R. McKinney,
“IR Spectroscopic Characteristics of a Cell Cycle and Cell Death Probed by
Synchrotron Radiation Based Fourier Transform IR Spectromicroscopy,”
Biopolymers (Biospectroscopy), 57:329–335, 2000.
3. P. Lasch, M. Boese, A. Pacifico, and M. Diem, FT-IR Spectroscopic Investigations of
Single Cells on the Subcellular Level,” Vibrational Spectroscopy, 28:147–157, 2002.
4. P. Lasch, A. Pacifico, and M. Diem, “Spatially Resolved IR Microspectroscopy
of Single Cells.” Biopolymers (Biospectroscopy), 67:335–338, 2002.
5. D. Yang, D. J. Castro, I. E. El-Sayed, M. A. El-Sayed, R. E. Saxton, and N. Y.
Zhang, “A Fourier-Transform Infrared Spectroscopic Comparison of Cultured
Human Fibroblast and Fibrosarcoma Cells: A New Method for Detection of
Malignancies,” Journal of Clinical Laser Medicine & Surgery, 13:55–59, 1995.