Arrangement for separating excitation light and emission light in a microscope

A fluorescence microscope, comprising:

• radiation means for emitting excitation light for irradiating a sample, whereby the sample emits excitation and wavelength-shifted fluorescence light, the radiation means comprising a plurality of light sources which provide excitation light of different wavelengths,
• detection means for detecting wavelength-shifted fluorescence light emitted by the sample,
• microscope optics means for directing excitation light from the radiation means towards the sample and for directing the excitation and the wavelength-shifted fluorescence light emitted by the sample back in the direction of the radiation means and the detection means,
• a plurality of acousto-optical means which are arranged on a common optical axis for individually feeding the excitation light from the light sources into the microscope optics means by diffraction of the excitation light, for independently regulating an excitation intensity of each of the light sources,
• and for separating excitation light and wavelength-shifted fluorescence light emitted by the sample by diffraction of the excitation light, the acousto-optical means being so positioned between the radiation means and the microscope optics means that only diffracted excitation light is introduced into the microscope optics means, wherein:
• in the direction of the microscope optics means as the acousto-optical means are firstly provided an AOM and then an AOTF,
• excitation light emitted by the sample is deflected by diffraction in the direction of the radiation means by the acousto-optical means, and
• wavelength-shifted fluorescence light emitted by the sample is transmitted undiffracted through the acousto-optical means and is thereby spatially separated from excitation light emitted by the sample,
• the detection means is so positioned with respect to the acousto-optical means that wavelength-shifted fluorescence light transmitted undiffracted through the acousto-optical means is detected by the detection means, and
• the acousto-optical means is so positioned with respect to the microscope optics means and the detection means that only undiffracted light is detected by the detection means, and further comprising:
• filter means for selectively detecting wavelength-shifted fluorescence light in the detection means positioned between the acousto-optical means and the detection means.

A device for feeding light into a microscope beam path and for detecting emission light emitted by a sample, comprising:

• a plurality of light sources which emit excitation light of different wavelengths, which excitation light is irradiated through the microscope beam path onto a sample, whereby the sample emits excitation and wavelength-shifted emission light which is directed back along the microscope beam path,
• detection means for detecting the wavelength-shifted emission light emitted by the sample, and
• a plurality of light diffracting means located on a common optical axis for individually feeding the excitation light of the plurality of light sources into the common optical axis by diffraction of the excitation light, for independently regulating an excitation intensity of each of the light sources, and for separating excitation light and wavelength-shifted emission light emitted by the sample by diffracting the excitation light and transmitting undiffracted the wavelength-shifted emission light, and wherein:
• at least one of the light diffracting means is associated with each light source and the different wavelengths by diffraction in the light diffracting means are simultaneously or individually fed into the common optical axis and are combined in the common optical axis,
• the detection means is so positioned with respect to the light diffracting means that only light transmitted undiffracted through the light diffracting elements is detected by the detection means, wherein the light diffracting means are chosen from the group consisting of an AOTF and an AOM, and wherein, in the direction of the microscope optics beam path, firstly an AOM and then at least one AOTF are arranged.