Thursday, 1 October 2015

LAB 1: PRINCIPLES AND USE OF MICROSCOPE



1.1 Setting up and using the microscope

Introduction
There are certain living organisms that are too small to be seen with our naked eye, such as microorganisms. So, the microscope is used to create an enlarged view or magnified visual of the microorganisms. There are two common types of microscope, light microscope and electron microscope. The light microscope is an instrument use visible light and magnifying lenses to view the microorganisms. It will provide visible details of the specimen to human eyes. It produces a better resolution than eyes, means that it able to distinguish two very small and close spaced objects as separate entities.

The part of microscope

The eyepiece tube receives the light that passes through the objective lens then redirects it to the eyepiece lens. The eyepiece lens have the magnification power of 10x.The objective lens of microscope provide four magnifications power, which are 4x, 10x, 40x and 100x. In order to produce a magnified primary image, the objective lens focuses the light beam which passed through the specimen. 
The stage of microscope is a place to put the slide of specimen. On the stage, there is a pair of stage clips that used to keep the slide in place. The diaphragm is used to adjust the total light that will enter the condenser; it has different size of holes and is used to control the intensity of light that will be directed onto the slide. The light source of microscope is come from the light source bulb which located at the base of microscope. 

The coarse adjustment knob and fine adjustment knob are important in the focusing of the image of specimen. When these knobs are adjusted, the stage will be raise or lower. The sequence of using the knobs is coarse adjustment knob followed by the fine adjustment knob. The purpose of the condenser lens is to focus the light onto the specimen.

Magnification and resolution
Resolving power of microscope is the ability to distinguish two very close distance and small objects as the different entities. Magnification is an apparent enlargement in size of an object so the object can be seen clearer and more details can be seen. By multiplying the objective lens' power and the eyepiece lens' power, the total magnification of the image of the specimen can be seen.
There are four magnifications given by microscope:

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


Objectives
To learn the correct and accurate way to use a bright-field microscope
To improve the knowledge regarding the magnification and resolution of the microscope
To provide more experience in the use of microscope

Materials and Reagents
Cover-slip, immersion oil, marker pen and microscope slide 

Procedure
(Refer to lab manual)

Results
Bacteria types: Cocci, Bacilli, Spirilli

 40 x magnifications
 100 x magnifications

 400 x magnifications
1000 x magnifications (oil immersion)


Discussion
There are three basic shapes of bacteria: coccus, bacillus, and spiral. Cocci may be spherical, oval, elongated, or flattened. They can exist in single coccus, pairs (diplococci), in chains (streptococci), in group of four (tetrad), in cube (sarcinae) and in clusters (staphylococci). Cocci do not dry out quickly and retain water, but they absorb nutrients slowly. Bacillus is rod-shaped and has four types: single bacillus, diplobacilli, strepobacilli, and coccobacillus. Bacillus has a large surface area that able to absorb nutrients, but they might dry out easily. Example for bacillus bacteria is Escherichia coli. Spiral bacteria have twists in three types: Vibrios (curved), Spirilla (helical) and Spirochetes (thin). Spirilla have flagella at both ends that allow them to move like a corkscrew, and capable to move faster than other bacteria. Their rigid cell wall helps to maintain their shape.


Conclusion
The experiment 1.1had showed the accurate way of using light microscope. We learn the components of microscope and their functions. Through this experiment, we learn the correct procedure to use the microscope to obtain a clear image of cells and microorganisms.


References
http://classes.midlandstech.edu/carterp/courses/bio225/chap04/lecture2.htm




1.2 Examination of cells

Introduction
Oil immersion technique is used to enhance the resolving power of microscope. The bacteria observed with the oil immersion objective are finer in resolution and have a brighter image. Wet mount method is used due to its quick preparation, simple steps and possible to observe moving and living organisms. 

Objectives
To learn how to make the wet mount slide
To obtain more experiences in using the microscope

Materials and reagents
Culture, immersion oil, lens tissue, a microscope slide containing stained microorganisms, inoculating loop, Bunsen burner, slide and cover slip

Lactobacillus fermentum


Procedure
(Refer to lab manual)

Results
Bacteria type: Lactobacillus fermentum
Shape: Rod
Size: Tiny
Colour: Transparent white



 1000 x magnifications (oil immersion)



Discussion
Lactobacillus sp is a rod-shaped, gram-positive and non-spore-forming bacterium belongs to family LactobacillaceaeLactobacillus sp has the ability to produce lactic acid as a by-product of glucose metabolism. Lactobacillus sp is normally non motile and can survive in both aerobic and anaerobic environments. Although they require rich media to grow, they are ubiquitous and may survive in anywhere contains carbohydrates, including respiratory, gastrointestinal, and urogenital tract of human and animals. In daily life, Lactobacillus sp is useful in the production of cheese and yogurt; in manufacturing of pickles and beverages like wine and juices. In medicine field, Lactobacillus sp is used for treating diarrhea and digestion problems such as irritable bowel syndrome (IBS) and colic in babies.


Conclusion
In the experiment 1.2, we observe the behaviour of Lactobacillus sp by using the oil immersion objective lens which is in high power. Besides, wet mount method is used to see the nature state of the bacteria. The technique of using microscope in experiment 1.1 helps us to use the microscope correctly to obtain a clear image of Lactobacillus sp. 
 

References
 

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