System and method for confocal raman-spectroscopic measurements of biological samples
Disclosed herein is a system, especially a microscope system, for confocal Raman-spectroscopic measurements. The system is configured to detect minute sample quantities of microbes in a sample chamber with a lid, whereas the system is configured such that a comparably large working distance between the aim lens and the sample has been sustained such that a dimension can be performed through the lid of the sample chamber. This allows for measuring and nurturing the sample at the exact same container.
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Several methods are known from the state of their art for detection and identification of microorganisms. Optical methods are particularly popular amongst the various detection methods due to their high speed and the minimal prep efforts theyrequire.
1 approach to discover and identify microorganisms is based on the test of inelastically scattered light from the specimen. This so-called Raman scattering can be used to discover but also to identify materials, molecules and the like based ontheir particular Raman spectrum. The Raman spectrum is reached by spectrally resolving the inelastically scattered light in the sample. Then, the measured spectrum is evaluated by fitting it with the Raman spectra of known substances ormicroorganisms.
In comparison to fluorescence-based detection, Raman-spectroscopic detection has the advantage which no labelling of the sample is required and the specificity of the Raman spectra connected to the several germs is more pronouncedthan the specificity of the associated (car -) fluorescence spectra.
As the Raman signal is generally orders of magnitude smaller than a fluorescence signal and as almost biological samples exhibit some degree of luminescence, such as for example fluorescence, the detection and separation of the Raman signal iscomparably demanding. Also, maintaining a sufficiently superior signal quality in Raman spectroscopic measurements becomes even more difficult once the quantity of the sample is small.
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