Abstract
The advantages of using a telecentric imaging system in digital holographic microscopy (DHM) to study biological specimens are highlighted. To this end, the performances of nontelecentric DHM and telecentric DHM are evaluated from the quantitative phase imaging (QPI) point of view. The evaluated stability of the microscope allows single-shot QPI in DHM by using telecentric imaging systems. Quantitative phase maps of a section of the head of the drosophila melanogaster fly and of red blood cells are obtained via single-shot DHM with no numerical postprocessing. With these maps we show that the use of telecentric DHM provides larger field of view for a given magnification and permits more accurate QPI measurements with less number of computational operations.
Highlights
Label-free quantitative phase imaging (QPI) of transparent microscopic specimens is the distinctive hallmark of digital holographic microscopy (DHM)
The ability of performing QPI has been extended to other microscopy methods like spatial light interference microscopy or spatial light interference tomography;[25,26] similar applications have been reported by the use of these imaging approaches
These phase changes encode the information about the refractive index and thickness of the specimen; their accurate measurement is determinant in the different applications where DHM can be utilized
Summary
Label-free quantitative phase imaging (QPI) of transparent microscopic specimens is the distinctive hallmark of digital holographic microscopy (DHM). In DHM, the fringe pattern recorded after the interference between a reference wave and the image of the wave diffracted by the microscopic sample allows the retrieval of the phase changes introduced by the sample These phase changes encode the information about the refractive index and thickness of the specimen; their accurate measurement is determinant in the different applications where DHM can be utilized. QPI-DHM working in the telecentric mode can provide the biomedical community with high-resolution, nondistorted phase images, which can be very useful for the diagnosis or screening of different diseases; biologist can be provided with trustable tool for specimen identification To this end, in this work, the performances of telecentric DHM and nontelecentric DHM are compared in terms of accuracy of the QPI measurements carried out on biological specimens with very different sizes. The obtained experimental results demonstrate the superiority of the telecentric mode for the study of biological samples
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