Confocal light scattering and absorption spectroscopic microscopy

We have developed a novel optical method for observing submicron intracellular structures in living cells which is called confocal light absorption and scattering spectroscopic (CLASS) microscopy. It combines confocal microscopy, a well-established high-resolution microscopic technique, with light scattering spectroscopy (LSS). CLASS microscopy requires no exogenous labels and is capable of imaging and continuously monitoring individual viable cells, enabling the observation of cell and organelle functioning at scales on the order of 100 nm. In addition, it provides not only size information but also information about the biochemical and physical properties of the cell.

[1]  Michael B. Wallace,et al.  Observation of periodic fine structure in reflectance from biological tissue: A new technique for measuring nuclear size distribution , 1998 .

[2]  Michael S. Feld,et al.  Polarized light scattering spectroscopy for quantitative measurement of epithelial cellular structures in situ , 1999 .

[3]  J. F. Aguilar,et al.  Imaging of spheres and surface profiling by confocal microscopy. , 2000, Applied optics.

[4]  Milind Rajadhyaksha,et al.  Confocal reflectance theta line scanning microscope for imaging human skin in vivo. , 2006, Optics letters.

[5]  S. Shapshay,et al.  Detection of preinvasive cancer cells , 2000, Nature.

[6]  Irving Itzkan,et al.  Confocal light absorption and scattering spectroscopic microscopy monitors organelles in live cells with no exogenous labels , 2007, Proceedings of the National Academy of Sciences.

[7]  M Fitzmaurice,et al.  Endoscopic detection of dysplasia in patients with Barrett's esophagus using light-scattering spectroscopy. , 2000, Gastroenterology.

[8]  I. Craig,et al.  Inverse Problems in Astronomy, A guide to inversion strategies for remotely sensed data , 1986 .

[9]  J. Fujimoto Optical coherence tomography for ultrahigh resolution in vivo imaging , 2003, Nature Biotechnology.

[10]  J. Mourant,et al.  Ultraviolet and visible spectroscopies for tissue diagnostics: fluorescence spectroscopy and elastic-scattering spectroscopy. , 1997, Physics in medicine and biology.

[11]  J. Lichtman,et al.  Optical sectioning microscopy , 2005, Nature Methods.

[12]  Irving Itzkan,et al.  Confocal light absorption and scattering spectroscopic microscopy. , 2007, Applied optics.

[13]  T Wilson,et al.  Imaging of spheres with the confocal scanning optical microscope. , 1996, Optics letters.

[14]  J. Lippincott-Schwartz,et al.  Development and Use of Fluorescent Protein Markers in Living Cells , 2003, Science.

[15]  Nathan C Shaner,et al.  A guide to choosing fluorescent proteins , 2005, Nature Methods.

[16]  W. Webb,et al.  Nonlinear magic: multiphoton microscopy in the biosciences , 2003, Nature Biotechnology.