Line-Scan Raman Microspectrometry for Biological Applications

Abstract

A high-resolution confocal line-scan Raman microscope was developed for the study of biological samples such as cells and chromosomes. With the illumination of a line on the sample, all the spectral information of the line is recorded. The line-scan microscope was attained by the introduction of one scanning mirror, a slit, and two cylindrical lenses in a standard confocal Raman microscope. The resolution, for an ideal sample, is 0.5 μm in the lateral direction and 3.5 μm in the axial direction. Line-scan experiments were performed to demonstrate the applicability of the instrument for biological samples. Line-scan images of calcium phosphate bone implant coatings and of the banding pattern of polytene chromosomes are presented.

Raman spectroscopy and microscopy of individual cells and cellular components

“…This major drawback is the main driving force for the development of CARS microscopy, which will be discussed in the next section. By employing alternative approaches, for example, by modifying the sample illumination to a line-scan approach [36,37] or employing novel electron-multiplying CCD cameras, the imaging time for spontaneous Raman imaging can also be significantly reduced, making it still an attractive method for label-free cell imaging. Fig.…”

Spatial Distribution of Phosphate Species in Mature and Newly Generated Mammalian Bone by Hyperspectral Raman Imaging

“…For example, de Grauw et al have used Raman microscopy to achieve 0.5 m spatial resolution at 512 points along a line at the interface between crystalline calcium phosphate and a section of bone. 11 Dental adhesive diffusion into dentin has been imaged using Raman microspectroscopy. 12 Marcott et al have reported 4096 pixel (33 mϫ33 m pixels) infrared spectroscopic images of canine alveolar bone tissue.…”

Monitoring Degradation Process of PLGA/Cap Scaffolds Seeded With Mesenchymal Stem Cells in a Critical-Sized Defect in the Rabbit Femur using Raman Spectroscopy

“…1/cm, detecting phosphate as a main element of hydroxyapatite. Therefore it is interpreted as an indicator of mineralized bone tissue [12]. …”

“…Mechanical loading of scaffolds with calcium has been described to accelerate the PLGA degradation [9]. of molecular structures and has already been used to specify calciumphosphate(CaP)-coated polymers and BRM [10][11][12]. It was also used for in vivo examinations in an animal model [13].…”