How does confocal Raman microscopy work?
In confocal Raman microscopy, a Raman spectrometer is coupled to a standard optical microscope, which leads to a high magnification of the sample and subsequent Raman analysis. Combining Raman spectroscopy with confocal microscopy allows optimal spatial filtering for sample volume analysis.
What is spatial resolution in Raman spectroscopy?
For example, scattering of the laser/Raman photons and interaction with interfaces in the sample can reduce this resolution. Thus, typical Raman spatial resolution is often quoted as being in the order of 1 µm, whilst with ‘good’ samples spatial resolution approaching the diffraction limit can be achieved.
What is a confocal microscope used for?
As a distinctive feature, confocal microscopy enables the creation of sharp images of the exact plane of focus, without any disturbing fluorescent light from the background or other regions of the specimen. Therefore, structures within thicker objects can be conveniently visualized using confocal microscopy.
What does Raman spectroscopy measure?
Raman spectroscopy is an analytical technique where scattered light is used to measure the vibrational energy modes of a sample. Raman spectroscopy can provide both chemical and structural information, as well as the identification of substances through their characteristic Raman ‘fingerprint’.
Why to use a confocal microscope?
Confocal microscopy offers several advantages over conventional widefield optical microscopy, including the ability to control depth of field, elimination or reduction of background information away from the focal plane (that leads to image degradation), and the capability to collect serial optical sections from thick specimens. The basic key to the confocal approach is the use of spatial filtering techniques to eliminate out-of-focus light or glare in specimens whose thickness exceeds the
What does confocal mean exactly in confocal microscope?
The Confocal Principle and Microscope Design ” Confocal ” is defined as “having the same focus.” What this means in the microscope is that the final image has the same focus as or the focus corresponds to the point of focus in the object. The object and its image are “confocal.”
What is the spatial resolution of a Raman microscope?
The spatial resolution of the Raman microscope is about 300 nm at the theoretical limit when using a 532 nm laser and a 0.90 NA objective lens, so the scanning accuracy (100 nm) of the motorized stage can be said to be sufficient.
What is confocal imaging use for?
Confocal microscopy, most frequently confocal laser scanning microscopy (CLSM) or laser confocal scanning microscopy (LCSM), is an optical imaging technique for increasing optical resolution and contrast of a micrograph by means of using a spatial pinhole to block out-of-focus light in image formation.
