International Journal of Scientific Research Engineering & Technology (IJSRET)

Volume 2 Issue 12 pp 835-836

March 2014

www.ijsret.org

ISSN 2278 0882

Digital Holographic Microscopy for high Resolution

Paresh V. Modh
R.R. Mehta collage of science
C.L. Parikh collage of commerce, Palanpur, Gujarat, India

ABSTRACT:
Digital Holographic Microscopy (DHM) provides
the quantitative distribution of the optical path
created by transparent. We used the reference
beam in DHM at an offset angle. Images of object
are generally obtained using photographic
method. The use of angular spectrum method is
also helpful to solve number of problems related
to microscopic.
KEY WORDS:
Two- step process, interference, reconstruction of
the image, digital holography, quantitative phase
imaging, biological specimens, Zero background,
and living cells.
Figure 1

I.
INTRODUCTION:
Holography is a two- step process. First step is
the recording of hologram where the object is
transformed into a photographic record and the
second step is the reconstruction in which the
hologram is transferred into image. A hologram is
the result of interference occurring between two
waves, an object beam which is the light scattered
off the object and a coherent back ground, the
reference beam, where the light is reaching the
photographic plate directly.
Recording of the Hologram:
In fig (1)There are two parts of the laser beam
<1> reference beam goes directly to the
photographic plate <2> The second beam of light
is directly on to The object to be photographed.
These two beams interfere and form interference
fringes. This photograph is known as holograph as
shown in fig (1) but this is negative plate.
In fig (2) reconstruction of the image must be
necessary in order to get real image. This type of
hologram is known as a transmission hologram
since The image is seen by looking Through it.

Figure 2
II.
IMPORTANCE:
In this letter, digital holographic microscopy
(DHM) is very important to get high resolution,
high- fidelity image of quantitative phase contrast
microscopy. In DHM, interferometric technique is
used. DHM is very simple and easy to operate
technique based on fig. (1) and fig. (2) Digital

IJSRET @ 2014

International Journal of Scientific Research Engineering & Technology (IJSRET)

Volume 2 Issue 12 pp 835-836

March 2014

holography, an emergent imaging technique,

offers an excellent approach for quantitative phase
imaging.
III.
APPLICATION:
With the help of conventional bright field
microscope, we cannot examine biological
specimens, such as living cells and their
intercellular constituents, because of transparent.
So these difficulties are removed by DHM. With
the help of DHM technique, we can see the living
cells, even though they differ from their
surrounding in terms of absorbance and colour.
We can get absolute phase distribution images of
living neurons in a culture obtained by use of the
DHM with accuracy in The 2.60 range.
There are number of technique like dark field
microscopy, Zernike phase contrast and Nomarski
differential interference contrast microscopes. In
dark field microscopy, we consider only the
scattering centers and boundaries contribute to the
image single against a Zero background. The
phase variation is converted into amplitude
variation, in the phase contrast microscopy. In
differential interference contrast (DIC), the
interference of two sheared polarization
components result in images that have shadow
effect and this give three- dimensional perfection
of the object. But we cannot get quantitative phase
image with the help of above techniques.
Holographic technique became a practical
proposition only after the invention of lasers.
Leith and upatnicks prepared laser holograms for
the first time so in fig. (3), Digital holographic
microscopy, laser is used.
In fig (3), basic configuration of The DHM is
shown. In digital holography, it is possible to
focus numerically on different object planes
without using any optomechanical movement.
The DHM is basically a Mach- Zehnder
interferometer. In figure 3 Bs= beam splitter, S=
sample, C= condenser, MO= microscope
objective, O
= object wave, M= Mirror, R=
Reference wave.
A laser beam is divided by a BS. The sample (S)
is illuminated by one beam through a condenser

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ISSN 2278 0882

(C). When the transmitted light by M is passing

through MO, then it is converted into object wave.
Here reference wave and object wave both are
interfere to each other. Holograms are recorded on
a standard monochrome CCD video camera.
Quantitative phase images are obtained by the
DHM according to an original procedure using a
single recorded hologram.

Figure 3
IV.
CONCLUSION:
We have demonstrated that a DHM is an efficient
and easy- to operate interferometric technique.
It is used biological environment to visualize
living cells in a culture. Holographic phase
images are obtained with 0.9m diffraction
limited lateral resolution. In DHM photographic
plate, each contains information about the entire
object form even a small part of the hologram the
entire image can be reconstructed if only with a
reduced clarity and definition of the image.
REFERENCES:
(1) M.K.
Kim,
Tomographic
Threedimensional imaging of a biological
specimen using Wavelength- scanning
digital interference holography.
(2) J.W. Goodman, introduction to Fourier
optics.
(3) T.M. Kreis, M. Adams and W.P.O.
Jueptner, Methods of digital holography.
(4) A
text
book
of
optics,
N.
subrahmanyamBrijlal, M.N. Avadhanulu.
(5) U. Schnar and W. P. Jueptner, Digital
Holography.

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