Light raster microscope and its use

1. A laser point scanning microscope comprising:

spot illumination means for providing an illumination beam for the illumination of a sample in the form of points,

a wide-field illumination source for illuminating the sample with wide-field illumination,

scanning means for guiding the illumination beam in the form of points over the sample so as to scan the sample, and for achieving spatial resolution by sampling of the sample both with illumination in the form of points and with wide-field illumination,

spot detector means including at least two spectral channels and a confocal aperture and a detector unit associated with each of the spectral channels for imaging, via the scanning means, the sample area illuminated by the spot illumination means and by the wide-field illumination source onto the detector units of different spectral channels using the associated confocal apertures, and

control means for controlling the scanning means during the operation of the wide-field illumination source and for controlling the spot detector means for reading out the spectral channels of the spot detector means for obtaining an image of the sample subject to wide-field illumination.

2. Laser point scanning microscope according to claim 1 , wherein the wide-field illumination source realizes one of transmitted light illumination for transmission measurement and incident light illumination of the sample for fluorescence excitation.

3. Laser point scanning microscope according to claim 1 , wherein the control unit controls the spot illumination means and the wide-field illumination source simultaneously during operation.

4. Laser point scanning microscope according to claim 1 , wherein the spot detector means comprises at least one Nipkow disk and at least one matrix detector.

5. Laser point scanning microscope according to claim 1 , wherein the spot detector means comprises at least one slit diaphragm and at least one row detector.

6. Laser point scanning microscope according to claim 1 , wherein the wide-field illumination source comprises a condensing lens configured for connection with means for adjusting the illuminated area.

7. Laser point scanning microscope according to claim 1 , wherein the scanning means comprises a scanning objective which exhibits the point spot to which at least one pupil plane P 1 is assigned, the scanning objective being configured for connection with means for adjusting the illuminated area.

8. Process for laser point scanning microscopy using the laser point scanning microscope according to claim 1 , comprising the steps of:

(a) illuminating a sample with an illumination beam in the form of a point using the spot illumination means,

(b) subjecting the sample to wide field illumination using the wide-field illumination source, and

(c) using at least the spot detector means, generating an image of the sample by scanning and confocal imaging of the sample illuminated by the illumination beam and the area of the sample illuminated by the wide-field illumination.

9. Process according to claim 8 , wherein in step (b), the sample is illuminated by transmitted light with the wide-field illumination.

10. Process according to claim 8 , wherein in step (c), the confocal imaging is performed with spectral resolution.

11. Process according to claim 10 , wherein the wide-field illumination of step (b) and the point illumination of step (a) are performed simultaneously.

12. Process according to claim 8 , wherein in the confocal imaging of step (c), at least one Nipkow disk is used in combination with at least one matrix detector.

13. Process according to claim 8 , wherein in the confocal imaging of step (c), at least one slit diaphragm is used in combination with at least one row detector.

14. Process according to claim 8 , further comprising the step of using one of phase contrast, dark-field contrast, VAREL contrast, polarization contrast, and differential interference contrast, as a contrasting method.

15. Method for studying developmental processes using the process of claim 8 , comprising the further step of:

(d) analyzing dynamic processes in the sample in a range of one-tenth of a second up to 1 hour, using the image generated in step (c).

16. Method for studying internal cellular transport processes using the process of claim 8 , comprising the further step of:

(d) studying small motile structures with high speed, using the image generated in step (c).

17. Method for representing molecular and other subcellular interactions using the process of claim 8 , comprising the further step of:

(d) representing very small structures with high speed, with the use of indirect techniques for the resolution of submolecular structures, using the image generated in step (c).

18. Method for studying fast signal transmission processes using the process of claim 8 , comprising the further step of:

(d) studying neurophysiological processes with high temporal resolution in the muscle or nerve system, using the image generated in step (c).

19. Method for studying developmental processes, comprising the step of:

analyzing dynamic processes in a range of one-tenth of a second up to 1 hour, at the level of united cell structures and entire organisms, using the laser point scanning microscope according to claim 1 .

20. Method for studying internal cellular transport processes, comprising the step of:

studying small motile structures with high speed, using the laser point scanning microscope according to claim 1 .

21. Method for representing molecular and other subcellular interactions, comprising the step of:

representing very small structures with high speed with the use of indirect techniques for the resolution of submolecular structures, using the laser point scanning microscope according to claim 1 .

22. Method for studying fast signal transmission processes, comprising the step of:

studying neurophysiological processes with high temporal resolution in the muscle or nerve system, using the laser point scanning microscope according to claim 1 .