Introduction Microscope

In biology, there are lots of things that are interesting but too tiny to see well - sometimes to see at all - with the naked eye. A magnifying lens is good, but a series of lenses, each magnifying the image of the last, works better. That's a microscope. Anything that you want to look at with a microscope can be called your specimen, and the nature of your specimen may dictate what sort of microscope you need to see it with. If you are looking at the outside, you want a scope that scans the surface - a scanning microscope images light (or other radiation) reflected off the surface of a specimen. To look inside, you need to get light (or other radiation) to go through your specimen - it needs to be thin, semi-transparent, or both. A microscope that sees images passed through a specimen is a transmission microscope.

History
The first microscope to be developed was the optical microscope, although the original inventor is not easy to identify. Galileo is sometimes credited with inventing the first simple microscope in 1610. Evidence points to the first compound microscope appearing in the Netherlands in the 1620s, probably an invention of eyeglass makers there. Two eyeglass makers there are variously given credit: Hans Lippershey (who developed an early telescope) and Zacharias Janssen (also claimed as the inventor of the

telescope). Robert Hooke is also cited as a possible inventor of the compound microscope. There are other claims that the microscope and the telescope was invented by Roger Bacon in the 1200s. Giovanni Faber coined the name microscope for Galileo Galilei's compound microscope in 1625 (Galileo had called it the "occhiolino" or "little eye").

Types of microscopes
Electron microscopy

In the early 1900s a significant alternative to light microscopy was developed, using electrons rather than light to generate the image. Ernst Ruska started development of the first electron microscope in 1931 which was the transmission electron microscope (TEM). The transmission electron microscope works on the same principle as an optical microscope but uses electrons in the place of light and electromagnets in the place of glass lenses. Use of electrons instead of light allows a much higher resolution.
Light Microscopy

It was not until the 1660s and 1670s that the microscope was used extensively for research in Italy, The Netherlands and England. Marcelo Malpighi in Italy began the analysis of biological structures beginning with the lungs. Robert Hooke's Micrographia had a huge impact, largely because of its impressive illustrations. The greatest contribution came from Antonie van Leeuwenhoek who discovered red blood cells and spermatozoa and

helped popularise microscopy as a technique. On 9 October 1676, Van Leeuwenhoek reported the discovery of micro-organisms.[6] In 1893 August Khler developed a key technique for sample illumination, Khler illumination, which is central to modern light microscopy. This method of sample illumination gives rise to extremely even lighting and overcomes many limitations of older techniques of sample illumination. Further developments in sample illumination came from Fritz Zernike in 1953 and George Nomarski 1955 for their development of phase contrast and differential interference contrast illumination which allow imaging of transparent samples.

Types of Light Microscopy

Bright field microscope

is the simplest of all the optical microscopy illumination techniques. Sample illumination is transmitted (i.e., illuminated from below and observed from above) white light and contrast in the sample is caused by absorbance of some of the transmitted light in dense areas of the sample. Bright field microscopy is the simplest of a range of

techniques used for illumination of samples in light microscopes and its simplicity makes it a popular technique. The typical appearance of a bright field microscopy image is a dark sample on a bright background, hence the name.

Dark field microscopy

(dark ground microscopy) describes microscopy methods, in both light and electron microscopy, which exclude the unscattered beam from the image. As a result, the field around the specimen (i.e. where there is no specimen to scatter the beam) is generally dark.

Fluorescence Microscope

A fluorescence microscope is an optical microscope that uses fluorescence and phosphorescence instead of, or in addition to, reflection and absorption to study properties of organic or inorganic substances. The "fluorescence microscope" refers to any microscope that uses fluorescence to generate an image, whether it is a more simple set up like an epifluorescence microscope, or a more complicated design such as a confocal microscope, which uses optical sectioning to get better resolution of the fluorescent image.

Phase contrast microscopy

Phase contrast microscopy is an optical microscopy technique that converts phase shifts in light passing through a transparent specimen to brightness changes in the image. Phase shifts themselves are invisible, but become visible when shown as brightness variations.

Figure 1: The same cells imaged with traditional bright field microscopy (left) and with phase contrast microscopy (right). When light waves travels through a medium other than vacuum, interaction with the medium causes the wave amplitude and phase to change in a manner dependent on properties of the medium. Changes in amplitude

(brightness) arise from the scattering and absorption of light, which is often wavelength dependent and may give rise to colors. Photographic equipment and the human eye are only sensitive to amplitude variations. Without special arrangements, phase changes are therefore invisible. Yet, phase changes often carry important information.

Polarised Microscopy
Polarised light microscopy uses plane-polarised light to analyse structures that are birefringent; structures that have two different refractive indices at right angles to one another (e.g. cellulose microfibrils). Normal, unpolarised, light can be thought of as many sine waves, each oscillating at any one of an infinite number of orientations (planes) around the central axis. Plane-polarised light, produced by a polar, only oscillates in one plane because the polar only transmits light in that plane.

Principles
The magnification of small things is a necessary facet of biological research, but the fine detail in cells and in subcellular components requires that any imaging system be capable of providing spatial information across small distances. Resolution is defined as the ability to distinguish two very small and closely-spaced objects as separate entities. Resolution is best when the distance separating the two tiny objects is small. Resolution is determined by

certain physical parameters that include the wavelength of light, and the light-gathering power of the objective and condenser lenses. A simple mathematical equation defines the smallest distance (d min) separating the two very small objects:

dmin = 1.22 x wavelength / N.A. objective + N.A. condenser

This is the theoretical resolving power of a light microscope. In practice, specimen quality usually limits dmin to something greater than its theoretical lower limit. N.A. (Numerical Aperture) is a mathematical calculation of the light-gathering capabilities of a lens.

References
1. Microbiology - Krishna Prakashan Media, page 13 2. Albert Van Helden, Sven Dupr, Rob Van Gent, Huib Zuidervaart, The Origins of the Telescope, pages 32-36 3. Microbiology - Krishna Prakashan Media, page 13
4. William Godwin (1876). "Lives of the Necromancers". 5. Gould, Stephen Jay (2000). "Chapter 2: The Sharp-Eyed Lynx,

Outfoxed by Nature". The Lying Stones of Marrakech: Penultimate Reflections in Natural History. New York, N.Y: Harmony. ISBN 0224-05044-3.

6. Wootton, David (2006). Bad medicine: doctors doing harm since

Hippocrates. Oxford [Oxfordshire]: Oxford University Press. ISBN 019-280355-7.

7. Knoll,

Max

(1935).

"Aufladepotentiel

und

Sekundremission

elektronenbestrahlter Krper". Zeitschrift fr technische Physik 16: 467475.

8. Morita

S (2006). Roadmap of Scanning Probe Microscopy.

NanoScience and Technology. Berlin: Springer. ISBN 3-540-34314-8.

9.

^ Majumdar A (1999). "Scanning Thermal Microscopy". Annual Review of Materials Science 29: 50585.

Bibcode:1999AnRMS..29..505M. doi:10.1146/annurev.matsci.29.1.505.

INDEX

 Introduction of Microscope History of Microscope Types of Microscope 1. Electron Microscope 2. Light Microscope Types of Light Microscope 1. Bright Field Microscope 2. Dark Field Microscope 3. Fluorescence Microscope 4. Phase Contrast Microscope 5. Polarised Microscope Principle Reference

Acknowledgement

Preparing

a project of this nature is an arduous task and I was

fortunate enough to get support from large number of person . I wish to express my deep sense of gratitude to all those who generously helped in successful completion of this report by sharing their invaluable time and knowledge It is my proud and privileged to express my deep regard to Respected Dr. J.P.N . Pandey Principal of Govt. Girls P.G College of Excellence Sagar Dr. Neera Sahay H.O.D. Zoology Department undertake the project I feel extremely exhilarated to have completed this project under the able for allowing me to

and inspiring guidance

of Mr. Rakesh Kumar Saket he rendered me all

possible help and guidance while reviewing the manuscript in finalizing this report . I also extend my deep regards to my teachers , family members, friends and all those whose encouragement has infused courage in me to complete the work successfully.

Gulafsha Kassab B.Sc. Sem IIIrd

Certificate

The project report titled Microscope in Sagar city prepared by Gulafsha Kassab B.sc. (Biotech) IIIrd Sem. under the guidance and supervision of Mr. Rakesh Kumar Saket , for partial fulfillment of the Degree B Sc.

Signature of supervisor

Signature of Examiner

Signature of H.O.D.

Acknowledgement

Preparing

a project of this nature is an arduous task and I was

fortunate enough to get support from large number of person . I wish to express my deep sense of gratitude to all those who generously helped in successful completion of this report by sharing their invaluable time and knowledge

It is my proud and privileged to express my deep regard to Respected Dr. J.P.N . Pandey Principal of Govt. Girls P.G College of Excellence Sagar Dr. Neera Sahay H.O.D. Zoology Department undertake the project I feel extremely exhilarated to have completed this project under the able and inspiring guidance of Mr. Rakesh Kumar Saket he rendered me all for allowing me to

possible help and guidance while reviewing the manuscript in finalizing this report . I also extend my deep regards to my teachers , family members, friends and all those whose encouragement has infused courage in me to complete the work successfully.

Sameeksha Mishra B.Sc. Sem IIIrd

Certificate

The project report titled Microscope in Sagar city prepared by Sameeksha Mishra B.sc. (Biotech) IIIrd Sem. under the guidance and supervision of Mr. Rakesh Kumar Saket , for partial fulfillment of the Degree B Sc.

Signature of supervisor

Signature of Examiner

Signature of H.O.D.

References
1.

Southern, Edwin Mellor (5 November 1975). "Detection of specific sequences among DNA fragments separated by gel electrophoresis". Journal of Molecular Biology 98 (3): 503517. doi:10.1016/S00222836(75)80083-0. ISSN 0022-2836. PMID 1195397.

2. Towbin et al.; Staehelin, T; Gordon, J (1979). "Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications". PNAS 76 (9): 4350. doi:10.1073/pnas.76.9.4350. PMID 388439.
3.

Burnette, W. Neal (April 1981). "Western Blotting: Electrophoretic Transfer of Proteins from Sodium Dodecyl Sulfate-Polyacrylamide Gels to Unmodified Nitrocellulose and Radiographic Detection with Antibody and Radioiodinated Protein A". Analytical Biochemistry 112 (2): 195 203. doi:10.1016/0003-2697(81)90281-5. ISSN 0003-2697.

PMID 6266278.
4. Biochemistry 3rd Edition, Matthews, Van Holde et al, Addison Wesley

Publishing, pg 977