Increasing the acquisition speed in oblique plane microscopy via aliasing

Area of Science:

• Biophysics
• Microscopy
• Cell Biology

Background:

• Oblique plane microscopy (OPM), a type of light-sheet fluorescence microscopy (LSFM), offers rapid volumetric imaging.
• OPM traditionally requires small scanning steps to satisfy Nyquist sampling due to tilted optical transfer functions (OTFs), limiting acquisition speed and increasing sample exposure.
• The tilted OTF in OPM necessitates careful sampling to avoid aliasing and maintain spatial resolution.

Purpose of the Study:

• To investigate if controlled under-sampling in OPM can accelerate image acquisition without compromising spatial resolution.
• To develop a method for recovering aliased OPM data to minimize artifacts and improve imaging speed.
• To demonstrate the application of this accelerated OPM technique for subcellular imaging.

Main Methods:

• Developed and applied a controlled under-sampling strategy during OPM acquisition.
• Utilized image recovery algorithms to correct for aliasing introduced by under-sampling.
• Performed 3D imaging of subcellular structures, including mitochondria and the endoplasmic reticulum.

Main Results:

• Demonstrated that judicious under-sampling in OPM can be recovered without loss of spatial resolution.
• Achieved significant speed gains, ranging from 2-fold to 4-fold, depending on system optical parameters.
• Successfully applied the method for rapid subcellular 3D imaging.

Conclusions:

• Controlled under-sampling is a viable strategy to accelerate OPM acquisition.
• This method allows for faster 3D imaging of cellular components with minimal artifacts.
• The developed technique enhances the utility of OPM for high-speed biological imaging.