Multi-focal structured illumination microscopy systems and methods

Abstract

Various embodiments for a multi-focal selective illumination microscopy (SIM) system for generating multi-focal patterns of a sample are disclosed. The embodiments of the multi-focal SIM system perform a focusing, scaling and summing operation on each generated multi-focal pattern in a sequence of multi-focal patterns that completely scan the sample to produce a high resolution composite image.

What is claimed is: 
     
         1 . A microscopy system comprising:
 a light source for transmitting a single light beam;   a first spinning disk with converging microlens array for rotation in a first direction and positioned along an optic axis for splitting the single light beam into a plurality of light beams forming a multi-focal pattern;   a second spinning disk with a pinhole array for rotation in the first direction and positioned along the optic axis for blocking out-of-focus light beams of the plurality of light beams for each multi-focal pattern and allowing through in-focus light beams of the plurality of light beams to pass through the spinning disk with a pinhole array, and a third spinning disk with a diverging microlens array for rotation in the first direction and positioned along the optic axis, wherein the first spinning disk with converging microlens array, the second spinning disk with a pinhole array, and the third spinning disk with a diverging microlens array rotate in sync relative to each other.   
     
     
         2 . The microscopy system of  claim 1 , wherein the third spinning disk with diverging microlens array have microlenses with half the focal length of the microlenses embedded within the first spinning disk with converging microlens array. 
     
     
         3 . The microscopy system of  claim 1 , wherein third spinning disk with diverging microlens array has the same number of microlenses as the first spinning disk with converging microlens array. 
     
     
         4 . The microscopy system  claim 1 , wherein the microlenses of the third spinning disk with a diverging microlens array are spaced at the same spatial location as the first spinning disk with a converging microlens array. 
     
     
         5 . The microscopy system of  claim 3 , wherein the first spinning disk with a converging microlens array, the second spinning disk with a pinhole array, and the spinning disk with a diverging microlens array are spaced apart from each other by the difference of focal lengths of the first spinning disk with a converging microlens array, the second spinning disk with a pinhole array, and the spinning disk with a diverging microlens array such that a Galilean telescope with a magnification of one half is arranged. 
     
     
         6 . The microscopy system of  claim 1 , further comprising:
 a dichroic mirror positioned between the laser source and the first spinning disk with converging microlens array and the second spinning disk with the pinhole array for diverting the single light beam from the laser source to the first spinning disk with converging microlens array.   
     
     
         7 . The microscopy system of  claim 1 , further comprising:
 an objective lens positioned between the second spinning disk with the pinhole array and a sample for illuminating the sample with the in-focus light beams of the plurality of light beams to generate fluorescence emissions by the sample.   
     
     
         8 . The microscopy system of  claim 7 , wherein the objective lens focuses the fluorescence emissions generated by the sample through the third spinning disk with diverging microlens array such that a camera detects the fluorescence emissions. 
     
     
         9 . The microscopy system of  claim 8 , further comprising:
 a telescope arrangement positioned between the third spinning disk with diverging microlens array and the camera for focusing the fluorescence emissions onto the camera for detection.   
     
     
         10 . The microscopy system of  claim 7 , further comprising:
 a tube lens positioned between the second spinning disk with pinhole array and the objective for focusing the single light beam to illuminate the sample and then focus the fluorescence emissions emitted by the illuminated sample through the second spinning disk with pinhole array, the first spinning disk with converging microlens array and the third spinning disk with diverging microlens array before detection of the fluorescence emissions by a camera positioned in operative communication with the third spinning disk with diverging microlens array.