Difference between Phase Contrast Microscopy and Differential Interference Contrast Microscopy

Phase Contrast vs DIC

Phase Contrast and Differential Contrast Microscopes

Phase contrast microscopy and Differential Interference Contrast (DIC) microscopy are two advanced optical light microscopy techniques to produce high contrast images of unstained and living cells. Both the microscopes utilize various contrast enhancing techniques to produce high contrast images.

Phase contrast microscopy is an optical-microscopy technique developed by Frits Zernike in 1934 to produce high contrast images of unstained live specimens. The phase contrast microscopy works by converting the phase shifts of light passing through a transparent specimen to detectable brightness changes in the image.

Differential Interference Contrast (DIC) microscopy, also called as Nomarski Interference Contrast (NIC) Microscopy, was first invented by Georges Nomarski in 1952. DIC microscopy uses more sophisticated contrast enhancing techniques than phase contrast system. It works by separating a polarized light source into two orthogonally polarized mutually coherent parts which are spatially displaced at the sample plane, and recombined before the final image formation. DIC produce more pronounced contrast difference than phase contrast image.

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The present post discusses the similarities and differences between phase contrast microscopy and Differential Interference Contrast Microscopy with a Comparison Table.

Similarities between Phase Contrast Microscopy and DIC Microscopy

Ø  Both Phase Contrast and DIC microscopes produce high contrast images.

Ø  Both are light microscopes, uses the visible spectrum of light.

Ø  Both uses two light beams to generate the final image.

Ø  Both are used for visualizing the live cells and living events.

Ø  Staining is not required for both types of microscopes.

Ø  Both the microscopes follow the Abbe’s law (limit of resolution).

Difference between Phase Contrast Microscopy and DIC Microscopy

Sl. No.Phase Contrast Microscope (PCM)Differential Interference Contrast Microscope (DIC)
What is Phase ContrastDIC image Exampe
1Phase contrast microscopy was developed by Zernike in 1934DIC was developed by Nomarski in 1952
2Phase contrast microscope works by the detection of sharp changes in the refractive indexDIC microscope works by the detection of continuous change of refractive index
3Phase contrast microscopy utilizes two light beams without beam splittersDIC utilizes two light beams with beam splitters
Light Path of Phase Contrast MicroscopeLight path of DIC
4Produce image by the incomplete separation of direct and diffracted light raysProduce image by the complete separation of direct and diffracted light rays
5Does not use a polarizerUses a polarizer
6Optical system of phase contrast microscopy do not requires beam recombinessOptical system of DIC requires beam recombiners
7Uses annular diaphragm and phase plate to create contrastDoes not use annular diaphragm and phase plate
8Nomarski’s prism is not used in the optical pathNomarski’s prisms are used in the optical path
9Produce a ‘halo’ around edges of the imageNo ‘halo’ is produced around the edges on the image
How halo is produced in Phase contrastHow halo is avoided in DIC
10Images from the phase contrast microscopy only provide qualitative dataImages from the DIC can produce both qualitative and quantitative data
11No three dimensional effects on the imagesImages are with a pseudo three dimensional effect
Image in Phase contrast3D Effects in DIC
12Full numerical aperture of the condenser and objective lenses cannot be utilized because of the masking effect of phase plate used in the light pathDIC microscopes can utilize the full numerical aperture of objective and condenser lenses
13Images in phase contrast microscope are not sensitive to the orientation of specimenImages in the DIC are highly sensitive to the orientation of the specimen
14Axial resolution lowAxial resolution high
15Optical sectioning is not possibleOptical sectioning is possible due to the high axial resolution
16Cost of the microscope is moderateCost is very high



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