Multispectral in-line hologram reconstruction with aberration compensation applied to Gram-stained bacteria...

Area of Science:

• Optical microscopy
• Holography
• Image reconstruction

Background:

• Optical aberrations limit the accuracy of multispectral digital in-line holographic microscopy (DIHM).
• Reconstruction of transmittance, phase, and morphology is affected by these aberrations, impacting repeatability.

Purpose of the Study:

• To develop and validate a method for calibrating and correcting optical aberrations in DIHM.
• To improve the repeatability and accuracy of quantitative phase and transmittance reconstructions.

Main Methods:

• Model fitting calibration using transparent beads to estimate wavelength-dependent, position-variant aberrations.
• Regularized inverse problem approach (IPA) for joint reconstruction of multi-wavelength holograms.
• Incorporation of shift-variant chromatic and geometrical aberrations into the forward model.

Main Results:

• The methodology demonstrated repeatable evaluation of optical aberrations.
• Improved repeatability in the reconstruction of transmittance and phase information.
• Enhanced contrast in spectral signatures for improved object analysis.

Conclusions:

• The developed method effectively corrects for optical aberrations in DIHM.
• This approach enhances quantitative imaging, particularly beneficial for stained biological samples like bacteria.
• Improved reconstructions support applications in microscopy-based analysis and classification.