Light-field microscopy image capturing method and light-field microscope

1 . Light-field microscopy image capturing method, comprising:

providing an illumination radiation from at least one light source;

modulating the illumination radiation by virtue of adjusting the intensity distribution thereof;

generating a dynamic light sheet in a sample space, in which a sample to be imaged can be arranged, by virtue of the modulated illumination radiation being radiated into an entrance location in an entrance pupil of a common illumination and detection objective and a first angle of incidence of the modulated illumination radiation being altered multiple times;

wherein a layer of the sample is illuminated with said dynamic light sheet only thinly at a time, avoiding bleaching outside the light sheet;

scanning a region of the sample to be imaged using the dynamic light sheet, the dynamic light sheet being brought into a number of different positions;

using the common objective to collect a detection radiation coming from the sample space;

guiding the collected detection radiation along a detection beam path and generating a plurality of partial image representations in a detection plane by means of a microlens array;

detecting the partial image representations as respective image signals of an image recording and evaluating the captured image signals on the basis of their associated location information and angle information;

modulating the illumination radiation by virtue of the light source being controlled directly and an intensity of the illumination radiation provided by the light source being altered in controlled fashion over time, and

recording an image whenever the dynamic light sheet is brought into a position;

or

recording an image while the dynamic light sheet adopts different positions.

2 . Image capturing method according to claim 1 , wherein the illumination radiation is modulated while the latter is scanned along a scanning path transversely to its propagation direction in relation to the sample space, with the modulation and the change of the first angle of incidence of the illumination radiation being matched to one another.

3 . Image capturing method according to claim 2 , wherein a modulation function is determined on the basis of an intensity distribution, known in advance, of the illumination radiation at rest and said modulation function is retrievably stored, with locations along at least one section of the scanning path being assigned modulation factors by way of the modulation function, with a desired intensity profile being implemented over the relevant section of the scanning path in the case of the control-technical implementation of said modulation factors in respect of location-related intensities of the scanned illumination radiation.

4 . Image capturing method according to claim 2 wherein the intensity of the illumination radiation is time-modulated during a scan transversely to the propagation direction such that a dynamic light sheet that is structured over its width is generated.

5 . Image capturing method according to claim 4 , wherein unwanted background signals are reduced by means of a combination by calculation of at least two image recordings.

6 . Image capturing method according to claim 1 wherein the modulated illumination radiation is shaped into a line-shaped light distribution and the line-shaped light distribution is radiated into the entrance location and a first angle of incidence of the line-shaped light distribution is altered multiple times.

7 . Image capturing method according to claim 1 wherein the dynamic light sheet is directed into the sample space at an inclination angle greater than zero degrees with respect to the optical axis of the common objective by virtue of the entrance location in the entrance pupil being chosen away from the optical axis.

8 . Image capturing method according claim 6 , wherein the optical axis passes through the chosen entrance location and the structured light sheet is generated in at least one position parallel to the optical axis.