Microscope, method and computer program for obtaining quantitative phase images by means of digital holographic microscopy

A microscope for obtaining quantitative phase images by means of digital holographic microscopy, comprising:

• a coherent light source (1) and a coherent light beam splitter (3) arranged at the output thereof for generating an object beam (Lo) with which a sample is illuminated and a reference beam (Lr);
• recording means (12) recording a hologram of said sample from an interference pattern of said reference beam (Lr) and a beam (Lo′) generated in said sample by the transmission or reflection of said object beam (Lo) thereon;
• and
• an optical system including optical elements arranged spaced apart from one another forming:
• a main optical path between a point of entry of said object beam (Lo) and said recording means (12), including optical elements making up a telecentric afocal system;
• and
• a reference optical path arranged between a point of entry of said reference beam (Lr) and said recording means (12), and sharing part of the optical elements with said main optical path;
• wherein:
• that said recording means (12) record said hologram in the image plane of said optical system, and
• said reference beam (Lr) is plane forming an angle φ with the optical axis of the microscope such that the following equations are complied with:
• NA
• M
• ≤
• λ
• 2
• 
• (
• 2
• +
• 3
• )
• 
• Δ
• 
• 
• p
• 
• 
• and
• 
• 
• sin
• -
• 1
• 
• (
• 3
• 8
• 
• π
• 
• NA
• M
• )
• ≤
• φ
• where λ is the wavelength generated by the coherent light source (1).