Identification of micro biological

organisms using digital holography

by
Raja Chakraborty
Department of Physics
Roll No:1521PH01

Sample chalk out of the presentation

Introduction
Literature Survey & Applications of DH
Set-up study
Discussion
Our future objective
Conclusion
References

Introduction

What is holography?

Literature survey
Applications of
Digital Holography

Marine

Environmental

Particle related

Biological

Set-up study

Optical setup of the digital in-line holographic microscope

Reference: Applications of Holographic Microscopy in Life Sciences,

http://dx.doi.org/10.5772/56176

Discussion
Marine applications

Involves study of zooplankton.

Underwater biological applications.

Study of marine organisms & subsea particles.

Incorporates Holo-camera (e.g Holocams, Hodars).

Holocam system deployed in East Sound,

WA, 2013.

Holocam data acquisition system

Reference: http://www.wetlabs.com

Raw holograms of the particle field outside & inside a thin layer of Phytoplankton.
The large chainformed in the right panel is genius Ditylum.
Reference:http://www.wetlabs.com

Environmental applications

Applications related to ice particles.

Applications related to cloud particles.

Study of micro properties of ground based

water particles.

Study of raindrops.

Particle related applications

Study of geometry (e.g size, shape, orientation

etc.) of particles like water, ice.

Detection of trajectories of particles.

Study of the velocimetry of the particle.

Detection of particles.

Particle tracking

DH technology as a
unique method for
detection of particles.

Several advantages &

Drawbacks of Digital
Holography.

Great applications &

future prospects.

Reference: S. Murata, N. Yasuda / Optics &

Laser Technology 32 (2000) 567- 574.

An example of hologram pattern

numerically obtained for 50 small
particles.

Set-up to record particle field

Typical digital in-line set-up for recording of particle field.

References: Ohmi. K., P. P. Sanjeeb & Joshi. B, Digital holography based particle
velocimetry, 10th International Conference on Fluid Control, Measurement, &
visualization, August 17-21, 2009, Moscow, Russia.

Biological applications

Detection & study of micro-organisms.

Investigation about Red blood cell.

Making of affordable point of care devices.

Study of cellular refractive indices.

Study of macroscopic biological

specimens

Fig. A : Bright field image of the Head of Drosophila melanogaster obtained with
conventional compound light microscope.
Fig. B & C : Two DIH imaged of Fig. A by using same objective separated by a depth of
6.8 micrometer. Blue laser; 2-mm pinhole; pinholeobject distance, 6.3 mm; pinhole
CCD chip distance, 6.9 cm.
Reference: Xu et al. PNAS, 98, 2011, 11301-11305.

Study of Red Blood Cell

Principal means of
delivering oxygen to the
body tissues.
The cytoplasm of
erythrocytes is rich in
hemoglobin.
The size of mature human
RBCs are 68 m in
diameter.
Nearly half of the blood's
volume (40% to 45%) is red
blood cells.

Reference: Wikipedia

Fig 1. Recorded Hologram.

Fig 2(a) &(b) : Phase & amplitude

image respectively.

Fig 2. Reconstructed Hologram

(1024 x 1024) pixel.

References: Salih. A & Tahir. K, Digital

in-line holography for blood cells,Int. J.
Phys. Sci., 8, 2013, 68-72

Our future objective

To identify different micro-organisms.

To study some different characteristics (for biological

organisms) through optical experiment.

To get some practical result like trajectory of particles, time

evolution of some bacteria, study of refractive indices etc.

Metrological study of Digital Holography.

To investigate about the significance & use of DH in the field

of bio-medicine, pharmacology , physiology, etc.

Conclusions

Visible range lasers can not produce good submicron

resolution.[ Xu et al. PNAS, 98, 2011, 11303].

lens-free imaging can reduce the dimensional gap between lab

on a chip components and microscopy to provide a powerful
approach to a portable POC diagnostic. [Kim et al. Journal of
Laboratory Automation, 17, 2011, 43 49].

It has huge real life biomedical application & use in laser

microsurgery of cells.

DH has great significance in the study of the dynamics of Red

blood cells.

References
.

Amsler, P., Stetzer, O., Schnaiter, M., Hesse, E., Benz, O. S., Moehler, and Lohmann, U., Ice crystal habits
from cloud chamber studies obtained by in-line holographic microscopy related to depolarization
measurements, Appl. Opt. 48, 2009, 5811-5822.

Bexon, R., Gibbs, J., and Bishop, G. D. Automatic assessment of aerosol holograms, J. Aerosol. Sci. 7,
1976, 397-407.

Black, K. S., Sun, H. Y., Craig, G., Paterson, D. M., Watson, J., and Tolhurst, T., Incipient erosion of
biostabilized sediments examined using particle-field optical holography, Environ. Sci. Technol. 35, 22752281.

Borrmann, S. and Jaenicke, R., Application of micro holography for ground-based in situ measurements in
stratus cloud layers: a case study, J. Atmos. Ocean. Technol. 10, 1993, 277-293.

Borrmann, S., Jaenicke, R., Maser, R., and Arends, B., Instrument intercomparison study on cloud droplet
size distribution measurements: holography vs. laser optical particle counter, J. Atmos. Chem. 19, 1994, 253258.

Borrmann, S., Jaenicke, R. and Neumann, P., On spatial distributions and inter-droplet distances in stratus
clouds with in-line holography, Atmos. Res. 29, 1993, 229-245.

Brown, P. R. A., Use of holography for airborne cloud physics measurements, J. Atmos. Oceanic Tech. 6,
1989, 293-306

Caballero, D., J. A., Loomis, N., Li, W., Hu, Q., Milgram, J., Barbastathis, G., and Davis, C., Advances in plankton imaging
using digital holography, in Computational Optical Sensing and Imaging, Optical Society of America, 2007, paper DMB5.

Costello, D. K., Carder, K. L., Betzer, P. R., and Young, R.W., In situ holographic individual particle dynamics and ocean flux
measurements, Deep-Sea Imaging of Settling Particles: Applications for Research 36,
36, 1989, 1595-1605.

Fugal, J. P., Shaw, R. A., Saw, E. W., and Sergeyev, A. V., Airborne digital holographic system for cloud particle
measurements, Appl. Opt. 43,
43, 2004, 5987-5995.

Fugal, J. P., Schulz, T. J., and Shaw, R. A., Practical methods for automated reconstruction and characterization of particles in
digital inline holograms, Measure. Sc. Technol. 20,
20, 2009, 075501.

Hobson, P. R. and Watson, J., Accurate three-dimensional metrology of underwater objects using replayed real images from
in-line and off-axis holograms, Measure. Sc. Technol. 10,
10, 1999, 1153-1161.
Hobson, P. R., Watson, J., The principles and practice of holographic recording of plankton, J. Opt. A: Pure Appl. Opt. 4,
2002, 534-549.

Hobson, P. R., Krantz, E. P., Lampitt, R., Rogerson, A., and Watson. J., A preliminary study of the distribution of plankton
using hologrammetry, Opt. Laser Technol.29
Technol.29,, 1997, 25-33.

Kendall, L. C, Robert, G. S, and Peter, R. B., In situ holographic measurements of the sizes and settling rates of oceanic
particulates, J. Geophvs. Res. 87, 1982, 5681-5685.

Kaikkonen,V. A., Ekimov, D., and Mkynen, A. J., A holographic inline imaging system for meteorological applications,
Proc. IEEE Int.I2MTC, 2013, 1630-1635.

Thank you