Pupil filters for extending the field-of-view in light-sheet microscopy

Area of Science:

• Microscopy
• Optical Engineering
• Biophysics

Background:

• Light-sheet fluorescence microscopy (LSFM) offers advantages for biological imaging but is limited by a narrow field of view.
• Extending the field of view is crucial for imaging larger biological structures or populations of cells with LSFM.

Purpose of the Study:

• To develop and validate a method for extending the field of view in LSFM using pupil filters.
• To optimize the design and implementation of binary phase modulation pupil filters for enhanced imaging.

Main Methods:

• Pupil filters, implemented as binary phase modulation elements, were designed using numerical optimization.
• The optimized filter structures were experimentally fabricated, scaled, and aligned within the LSFM system.
• The effect of the pupil filters on the field of view was quantitatively assessed.

Main Results:

• Numerical optimization successfully determined optimal pupil filter structures.
• Experimental implementation resulted in a significant and practical extension of the microscope's field of view.
• The approach demonstrated the feasibility of wide-field imaging with LSFM.

Conclusions:

• Binary phase modulation pupil filters are an effective strategy for extending the field of view in LSFM.
• The combination of numerical optimization and experimental validation provides a robust method for filter implementation.
• This technique represents a valuable advancement towards achieving wide-field light-sheet microscopy.