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Near-field microscopy and fluorescence spectroscopy: application to chromosomes labelled with different fluorophores
- Source :
- Journal of Microscopy, Journal of Microscopy, Wiley, 2010, 238 (1), pp.36-43. ⟨10.1111/j.1365-2818.2009.03326.x⟩
- Publication Year :
- 2010
- Publisher :
- Wiley, 2010.
-
Abstract
- We have coupled a spectrophotometer with a scanning near-field optical microscope to obtain, with a single scan, simultaneously scanning near-field optical microscope fluorescence images at different wavelengths as well as topography and transmission images. Extraction of the fluorescence spectra enabled us to decompose the different wavelengths of the fluorescence signals which normally overlap. We thus obtained images of the different fluorescence emissions of acridine orange bound to single or double stranded nucleic acids in human metaphase chromosomes before and after DNAse I or RNAse A treatment. The analysis of these images allowed us to visualize some specific chromatin areas where RNA is associated with DNA showing that such a technique could be used to identify multiple components within a cell.
- Subjects :
- Méthodes et systèmes en IRM et optique
Histology
Materials science
genetic structures
Physics::Optics
02 engineering and technology
Fluorescence spectroscopy
Pathology and Forensic Medicine
03 medical and health sciences
Optics
Microscopy
Image Processing, Computer-Assisted
categₛt2i
[INFO.INFO-IM]Computer Science [cs]/Medical Imaging
Fluorescence microscope
Chromosomes, Human
Humans
ComputingMilieux_MISCELLANEOUS
Fluorescent Dyes
030304 developmental biology
Fluorescence loss in photobleaching
[PHYS.PHYS.PHYS-OPTICS]Physics [physics]/Physics [physics]/Optics [physics.optics]
0303 health sciences
Total internal reflection fluorescence microscope
Staining and Labeling
business.industry
reseauₙational
021001 nanoscience & nanotechnology
imagerie_RMNₒptique
Chromatin
Spectrometry, Fluorescence
Light sheet fluorescence microscopy
Near-field scanning optical microscope
Fluorescence cross-correlation spectroscopy
sense organs
0210 nano-technology
business
Subjects
Details
- ISSN :
- 13652818 and 00222720
- Volume :
- 238
- Database :
- OpenAIRE
- Journal :
- Journal of Microscopy
- Accession number :
- edsair.doi.dedup.....f8af310325a60d548ceabacedc6e99cb