Sunday, October 4, 2015

Lab 1: Principles and Use of Microscope

1.1 Setting up and using the microscope

Introduction:
A microscope is an instrument used to see objects that are invisible to the naked eye. The science of investigating small objects using such an instrument is called microscopy. Microscopic means invisible to the eye unless aided by a microscope. There are many types of microscopes. The most common use is the optical microscope (light microscope), which uses visible light and a system of lenses to magnify images of small samples.

The diagram below shows the parts of the microscope:


STRUCTURAL COMPONENTS OF A MICROSCOPE
The three basic, structural components of a compound microscope are the head, base and arm.
•           Head/Body: Houses the optical parts in the upper part of the microscope
•           Base: Supports the microscope and houses the illuminator
•           Arm: Connects to the base and supports the microscope head. It is also used to carry the   microscope.

OPTICAL COMPONENTS OF A MICROSCOPE
There are two optical systems in a compound microscope: Eyepiece Lenses and Objective Lenses:
·         Eyepiece or Ocular: The lens at the top that you look through. Typically, standard eyepieces have a magnifying power of 10x.
·         Eyepiece Tube: Holds the eyepieces in place above the objective lens. Binocular microscope heads typically incorporate a diopter adjustment ring that allows for the possible inconsistencies of our eyesight in one or both eyes. Binocular microscopes also swivel (Interpupillary Adjustment) to allow for different distances between the eyes of different individuals.
·         Objective Lenses: The primary optical lenses on a microscope. They range from 4x-100x and typically, include, three, four or five on lens on most microscopes.
·         Nosepiece: Holds two or more objective lenses and can be rotated to easily change power.
·         Coarse focus knob: It is used to focus on the specimen. It may move either the stage or the upper part of the microscope in a relative up and down motion.  
·         Fine Focus knobs: It is the smaller round knob on the side of the microscope used to fine-tune the focus of the specimen after using the coarse adjustment knob.
·         Stage Clips: Used when there is no mechanical stage. The viewer is required to move the slide manually to view different sections of the specimen.
·         Aperture: The hole in the stage through which the base (transmitted) light reaches the stage.
·         Illuminator: The light source for a microscope, typically located in the base of the microscope.
·         Iris Diaphragm: Controls the amount of light reaching the specimen. It is located above the condenser and below the stage.
·         Condenser Focus Knob: Moves the condenser up or down to control the lighting focus on the specimen.
Magnification and resolution:
            Magnification is the ability to make small objects seem larger, such as making a microscopic organism visible. There are four magnifications in the microscope used:
4x objective X 10x eyepiece = 40x magnification
10x objective X 10x eyepiece = 100x magnification
40x objective X 10x eyepiece = 400x magnification
100x objective X 10x eyepiece = 1000x magnification
            Resolution is the ability to distinguish two objects from each other. Not to be confused with magnification, microscope resolution is the shortest distance between two separate points in a microscope’s field of view that can still be distinguished as distinct entities.
Objective:
-To learn the proper way of handling and care of microscope.
-To understand the basic concept of magnification and resolution of a microscope.

Materials and reagents:
-Microscope slide and cover-slip

Procedure:
(Refer to the lab manual)

Results 

Typical Bacillus is observed under the microscope


40x magnification



 100x magnification


400x magnification



1000x maginification (oil immersion)


 Discussion

We observed the specimen from lowest magnification that is 40x to the highest magnification 1000x (oil immersion). The specimen that we observed is Typical Bacillus. Bacillus is a genus of Gram-positive which appeared pink in color when observed under the microscope. Bacillus is one of the best understood prokaryotes, in terms of molecular and cellular biology. The cell wall of Bacillus is a structure on the outside of the cell that forms the second barrier between the bacterium and the environment and at the same time maintains the rod shape and withstands the pressure generated by the cell’s turgor. The cells are straight, round-ended or square-ended rods. The size of Typical Bacillus is tiny and the surface as it was observed under the magnification of 1000x with oil immersion, it looked smooth and the texture is moist. Most species motile by peritrichous flagella. Many Bacillus species have little or no pathogenicity and are rarely associated with disease in humans or lower animals except Bacillus anthracis, Bacillus cereus and Bacillus subtilis. Some species are insect pathogens.

Conclusion

In conclusion, we have learnt the right way of handling the microscope with care and also how to observe a specimen under the microscope from the lowest magnification to the highest magnification using a correct way. We find that the higher the magnification, the clearer the image that we can see under the microscope.



1.2 Examination of cells


Introduction

Living microorganisms are very diverse in their type, size and shape but most importantly many of them are also colorless when viewed using the microscope. It is essential as a biology student that we can search and found easily the microorganisms that is used so that we can examine and learn from it.

As such, by using the wet mount methods which does not require any special equipment, not harmful when done properly and significantly quick and easy we can studies more on microorganism easier. The wet mount methods are a technique that enable us to experience the proper use of microscope and aseptically prepare a slide thus, we are learning in a safe environment.

Objective

-To provide an experience in the use of microscope
-To illustrate the diversity of cells and microorganisms

Materials and reagents

-Culture
-Immersion Oil
-Lens Tissue 
-A microscope slide containing stained microorganisms
-Inoculating loop
-Bunsen burner 
-Slide and coverslip

Result


Lactobacillus (1000x magnification with oil immersion)


Discussion

The wet mount methods enable us to observe the Lactobacillus more clearly of the sizes and the shape. Oil immersion fills the space between the objective lens and specimen and matches the refractive index of the glass cover slip and glass objective lens. At a given focal length, greater numerical aperture can be achieved.
Lactobacillus is a genus of Gram-positive facultative anaerobic or rod-shaped bacteria. They are a major part of the lactic acid bacteria group. In humans they are part of the vaginal microbiota. Lactobacillus is a type of bacteria with multiple different species in the genus. Most Lactobacillus species in humans are considered harmless. Lactobacilli live in the urinary digestive and genital tracks of humans.Some Lactobacillus species are used as starter culture in industry for controlled fermentation in the production of yogurt, cheese, beer, wine etc.
In this experiment, aseptic techniques is important to ensure that the bacteria is not contaminated. It is also to protect the user from infection and to prevent the spread of microorganisms. Through this experiment, Lactobacillus appeared to be translucent because it was not stained.

Conclusion

The bacteria can be seen more clearly with the highest magnification under oil immersion. Added with wet mount method, the bacteria can be seen clearly in its natural state. Proper aseptic technique is important because some of the pathogenic agents can cause infection or serious illness to the user if they are not handled in a proper way. 

References
https://en.wikipedia.org/wiki/Microscope
http://www.microscope.com/education-center/microscopes-101/compound-microscope-parts/

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