Method for microscopy, and microscope

Abstract

A method for microscopy includes generating pulsed illuminating light including wavelengths in a spectral region. A detection spectral region within the spectral region is defined. Using a dynamically controllable mask, light components of the illuminating light that comprise wavelengths within the detection spectral region are influenced. A specimen is illuminated with the influenced illuminating light. Detection light proceeding from the specimen within the detection spectral region is detected.

Description

CROSS REFERENCE TO RELATED APPLICATIONS [0001] This is a continuation of application Ser. No. 10 / 601,804, filed Jun. 23, 2003, which claims priority to German patent application 102 28 374.5. The subject matter of both of these applications is hereby incorporated by reference herein.FIELD OF THE INVENTION [0002] The invention concerns a method for microscopy. The invention furthermore concerns a microscope and a confocal scanning microscope. BACKGROUND OF THE INVENTION [0003] For the investigation of biological specimens, it has been usual for some time to prepare the specimen using optical markers, in particular fluorescent dyes. Often, for example in the field of genetic research, several different fluorescent dyes are introduced into the specimen and become attached specifically to certain specimen constituents. From the fluorescence properties of the prepared specimen conclusions can be drawn, for example, as to the nature and composition of the specimen or the concentration of ...

AI Technical Summary

Benefits of technology

The invention provides a method for microscopy that allows for efficient exploitation of nonlinear processes with reduced specimen damage. The method involves generating pulsed illuminating light that includes wavelengths within a spectral region, influencing the light components within that region, and then illuminating a specimen with the influenced light. The detection light from the specimen is then detected within a detection spectral region that lies within the spectral region of the illuminating light. The invention also provides a microscope that reliably and efficiently allows investigation of a specimen using nonlinear processes with reduced specimen impact. The method and microscope make use of lower light intensities and can focus on a specimen volume to detect in practically background-free fashion the detection light produced by nonlinear processes. The power level and spectral distribution of the detection light are used for image production. The invention also provides a spectral filter that removes certain frequency regions from the spectrum of the illuminating light to create a spectral window within which detection light produced as a result of nonlinear processes can be detected in background-free fashion. The spectral filter can allow only light of the wavelengths of the detection spectral region to arrive at the detector. The invention also provides a scanning microscope that allows for efficient exploitation of nonlinear processes with reduced specimen damage.

Problems solved by technology

It must be noted, however, that illuminating light having at least two different illuminating light wavelengths, at high light power levels, is required for this method.

The aforesaid methods are disadvantageous in that high light power levels are necessary, resulting on the one hand in great demands on the light source and on the other hand in undesirable damage to the specimen, for example due to bleaching.

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