Page 8 - Vibrational Spectroscopic Imaging for Biomedical Applications
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2.3 Flow Cell for In Vivo IR Microspectroscopy
of Biological Samples . . . . . . . . . . . . . . . . . . . . 36
2.3.1 Flow Chamber Design . . . . . . . . . . . . 39
2.3.2 Mid-IR and Vis Measurements . . . . . 42
2.3.3 Viability Tests: PAM Fluorescence
Measurements . . . . . . . . . . . . . . . . . . . 44
2.3.4 Initial Flow Cell Measurements
with IRENI . . . . . . . . . . . . . . . . . . . . . . 47
2.4 Biomedical Application: Calcium-Containing
Crystals in Arthritic Cartilage . . . . . . . . . . . . . 48
2.4.1 Calcium-Containing Crystals
and Arthritis . . . . . . . . . . . . . . . . . . . . . 48
2.4.2 Current Methods of Crystal
Identification . . . . . . . . . . . . . . . . . . . . 49
2.4.3 Biologic Models of Calcium-
Containing Crystal Formation . . . . . . 49
2.4.4 Synchrotron-Based FTIR
Microspectroscopy Spectral Analysis
of Calcium-Containing Crystals . . . . 50
2.5 Future Directions: In Vivo Kinetics of
Pathological Mineralization and
Phytoplankton Adaptation . . . . . . . . . . . . . . . 55
Acknowledgments . . . . . . . . . . . . . . . . . . . . . . . . . . . . 55
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 56
3 Preparation of Tissues and Cells for Infrared and
Raman Spectroscopy and Imaging Ehsan Gazi and
Peter Gardner . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 59
3.1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . 59
3.2 Tissue Preparation . . . . . . . . . . . . . . . . . . . . . . . 61
3.2.1 Archived Tissue: Paraffi n Embedded
and Frozen Specimens . . . . . . . . . . . . 61
3.2.2 Preparation of Tissues for Diagnostic
Assessment Using FTIR and
Raman Microspectroscopy . . . . . . . . . 63
3.2.3 The Effects of Xylene on Fixed
Tissue and Deparaffi nization of
Paraffin-Embedded Tissue . . . . . . . . . 68
3.3 Cell Preparation . . . . . . . . . . . . . . . . . . . . . . . . . 71
3.3.1 Chemical Fixation for FTIR and
Raman Imaging . . . . . . . . . . . . . . . . . . 71
3.3.2 Sample Preparation for Biomechanistic
Studies . . . . . . . . . . . . . . . . . . . . . . . . . . 78
3.3.3 Growth Medium and Substrate Effects
on Spectroscopic Examination
of Cells . . . . . . . . . . . . . . . . . . . . . . . . . 80
