Bright field microscopy was used for stained samples; while Dark field and Phase contrast microscopy are used for the non-stained specimens which can e illuminated from the side and top (for specimen that are not suitable to be observed under dark field microscopy) respectively. Koehler illumination is done to create high contrast illumination and improve microscopic image outcome. The aim was the study was multi fold; generating microscopic specimen of CHOC cells in which we used observed with different microscopic visualization techniques as well as the different stages of mitosis.
Then after we examined the crucial relationship between structure and function. Finally, the most efficient way of cleaning your hands and prevent bacterial growth, using was analyzed. The CHOC cell samples were fixed; one with formaldehyde and another sample with maceration solution (ethanol/hydrochloric acid) and stained with origin solution . In order to evaluate the levels of hygiene resulting from different methods of washing one’s hands, colonies formed on agar plates with fingerprints after using : water, water with soap and 70% OTOH, were compared with one another and with unwashed hand colonies.
The structure-function relationship is important for the identification of cell types. By identifying the cell organelles, we determined the tissues in the pictures showed as liver, kidney, increase, plasma, or accessory testis cells (Datelined and Brim, 2013). 4. Discussion 4. 1 Microscopy As one of the three sections of the experiment, microscopic specimen of CHOC cells were generated and used to learn about microscopic visualization techniques as well as the different stages of mitosis examined cell division in CHOC cells.
The variation on the contrast techniques are based on differences with the source of the light for the light microscopy. With light microscopy the specimen is illuminated from below and thus requires stained specimen; this enhances visualization of the cell or certain cellular components under a gyroscope. The use of different stains enables one to preferentially stain certain cell components (such as a nucleus or a cell wall) or the entire cell. Most stains can be used on fixed, thus non-living cells, while only some can be used on living cells (Basic cell staining, 2013).
Dark field microscopy, where specimen is illuminated from the side, is ideal when dealing with specimen that are unstained, transparent and absorbing little to no light. However, most of the details in cells are undetectable in bright field microscopy because there is too little contrast between structures with similar transparency and there is inadequate natural pigmentation. Thus we use the great range in refractive index various organelles show, providing an opportunity to distinguish them.
By this a non-visible phase shift is transformed into differences in the amplitudes of light waves, which are visible for the human eye (Phase contrast microscopy, 2012). Comparing all three we found stained samples in phase contrast the best as we were able to detect most details including the cell cycle with it. In Interprets the cell is engaged in metabolic activity and preparation for mitosis (the next four phases in cell division). Chromosomes are not clearly seen in the clues, although the nucleolus could still be visible. The connectors is the major microcircuit organizing center in animals where a microcircuit radial array originates.
Whereas in Protease chromatin in the nucleus begins to condense and recognizable under light microscope as chromosomes. The nucleolus disintegrates and controller begin moving to opposite ends of the cell while fibers extend from the controverts forming the mitotic spindle. We found 22 % of the cells to be going through Protease, which is a lower number compared to the Telephone. This could have been due to the fact that the count was not add from the same area or the cells were fixated while one cycle was ending. Cells undergoing Protease and Interprets are difficult to differentiate as they are very similar.
However, in the cell cycle Interprets cells replicate the DNA and grow, but do not undergo mitosis until they get to Protease. In Interprets the nuclear envelope is present and the misconstrues are not organized in a mitotic spindle. Higher magnification and improved resolution , enabling observation of the detailed differences between both cells could improve the results. The nuclear membrane dissolves in Promethean; protein structures, constrictors, attach to the controverts where the misconstrues, spindle fibers, attach during cell division to pull the sister chromatics apart.
Whereas in Metastases the spindle fibers align the chromosomes in the center of the cell nucleus and the line is known as the metastases plate. The arrangement helps to ensure that in the process where the chromosomes are separated, each new nucleus will have one copy of each chromosome. The pair of chromosomes separate at the constrictors and move to opposite sides of the cell o in the stage referred to as Anapest. The separation motion is caused by a combination of eigenvector event along the spindle misconstrues and physical interaction of polar misconstrues.
The least prominent phase in the counted cells is Anapest, with 17%. This could have been due to the fact that the Anapest and Metastases were exchanged during Finally in Telephone chromatics arrive at opposite poles of cell, and new membranes are form around the daughter nuclei. This is when chromosomes and spindle fibers disperse and are no longer visible under the light microscope. The most prominent phase observed was Telephone , with 38%. Cytokines, partitioning of the cell, begins during this stage.
In the CHOC cells, woodiness results when a fiber ring around the center of the cell composed of a protein called acting contracts pinching the cell into two daughter cells, each with one nucleus (Cell cycle and mitosis, 2004). Cells during interprets are more flat and oval than those undergoing mitosis. While the cells in mitosis are more round yet during the different phases show varying cell organelle structure. Since cells are not stained, organelles are not visible, expect for the nucleus and cell membrane. The fixation of living cells and tissues is an immediate stop of the life processes taking place within and around them.
Fixation is a tool for preservation for a long time, otherwise it may cause disintegration of the structures. The first sample of the CHOC cells are treated with ethanol and hydrochloric acid. These alcohol/acid combination has significant effects on the cell and are used to prepare the cells for staining with origin. Ethanol is a non- additive precipitant fixative. Proteins are fixed by dehydration and precipitation, the degree to which this is done relies on the amount of water present and the solubility of the materials in the mixture.
Thus the Ethanol/HCI fixative is water ere or contain only minor amounts of water so that precipitation is a major effect. By disrupting the hydrophobic interactions that give many proteins their tertiary structure ethanol solubility the membrane. This precipitation and aggregation is a very different mechanism from the cross linking that occurs with the formaldehyde fixative. Ethanol causes considerable shrinkage and hardening of tissue during fixation. In addition to DNA denaturing, HCI alone is associated with tissue swelling; combining the two may result in better preservation of the cells.
We treated the unstained samples with formaldehyde. Formaldehyde reacts primarily with proteins; the allayed group can react with nitrogen and some other atoms of proteins. This will form a Methyl bridge is formed between two reactive atoms in proteins that are very close together. Other molecules (carbohydrates, lipids, nucleic acids) are thought to be trapped in a matrix of cross linked proteins, thereby forming a rigid assembly further conserving the cell (Cross linking fixatives, 2013 4. 2 Hygiene test Pathogens are frequently acquired on hands and hand washing is essential to remove them.
As the skin is dry, acidic and poor in nutrients its considered to be n inhospitable environment for most microorganisms. Microorganisms adapted to these conditions exist in stable populations known as resident or normal floury. While those acquired on the surface of the skin through contact with other people, objects or environment are known as transient skin flora. Transient microorganisms can usually be easily removed by briefly washing with soap and water. In the case of resident microorganisms , even though their numbers can be reduced by using soap or detergent they are not easily removed by it.
We expected that everybody working in a laboratory environment will bring bacteria onto the laboratory. The amount of microorganisms on the hand varies with the location; with thee density of microorganisms being the highest under the fingernails. Alcohol solutions (60 – 95 %) are quite microbial and active against both Gram-negative and Gram positive bacteria, fungi and most viruses. Furthermore, alcohol does appear to suppress bacterial growth on skin for a few hours after application and is effective against both resident and transient skin flora (Infection control in clinical practice, 2006, p. 57-159). According to the results from the hygiene test, the general pattern shows that washing with ethanol is better than with soap and water which is better than with just water. Washing by water itself would decrease the amount of colonies as some would be removed due to hydrophobic and/ or the pressure exerted by the water. Because soap is a salt, it partially separates into its component ions in water. The two ends of this ion behave in different fashions. When soap and water are mixed, soap molecules are uniformly dispersed.
This system is not a true solution , however, because the hydrocarbon portions of the soap’s ions are attracted to each other and form spherical aggregates known as micelles. When art is added to this system, it is taken into these micelles as tiny particles and rinsed away. In some cases however, the number of colonies increased after cleaning with soap or ethanol (Table 1 and Table 2 This could have been caused by closing the tap after with the hand to be tested or leaving the ethanol open for long which will make it evaporate thus less effective (personal communication with Dry.
Datelined, 2013-04-12). 4. 3 Electron microscopy identification One of the different aims of this experiment is that of the structure-function relationship ; the biological structure analysis better acquaints us with what ND/or how it works. After careful observation of the cell organelles, what they do and why they do it, we were able to determine the identity of each specific tissue. On ME picture #1 a tremendous amount of vesicles, multi vesicular bodies and Googol apparatus can be observed.
The accessory testes produce much- polysaccharides and fluids which are stored in vesicles and upon a signal are secreted to be added to the sperm cells, to form the semen (Datelined and Brim, 201 3). The vesicles are used to secrete proteins and other ,macromolecules to the outside of the cell while proteins are sorted and packaged by the Googol complex. Then after they fuse with the plasma membrane and discharge their contents to the exterior of the cell by the process of exoticism (Googol Apparatus and transport, 2010). Thus it fits the description of accessory testis tissue (Table 4).
The fact that there is no nucleus visible may be due to the fact that the cells were not shown as a whole (personal communication with Proof. Datelined, 12. 04. 2013). ME picture #2 shows a great deal of mitochondria alongside the microvolt and a capillary with basement membrane and poetess. The large amount of mitochondria found in kidney cells, are needed as re-absorption from the filtrate involves active transport. The microvolt cell lining in these cells increases the surface area for the absorption of materials from the filtrate.
The basement membrane is the floor of an epithelial membrane on which the basal cells rest. Poetess that wrap around the capillaries of the a network hat performs the first step of filtering blood. These all included in the ME picture fits the description of kidney tissue (The kidney, 2013). On ME picture #3 we observe quite a lot of genome granules and extensive RE (both smooth and rough) . The excessive presence of both endoplasmic testicle indicates that there has been major manufacture of macromolecules.
The rough RE is responsible in the production of membranes and secretors proteins; while the smooth RE has a wide range of functions including carbohydrate and lipid synthesis. The pancreas, a digestive organ, secrets digestive enzymes that are responsible for the breakdown of absorbed macromolecules. Digestive enzymes are released in inactive forms called genomes in order to prevent them from digesting the cells that produce them. On the basis of the functions, the ME picture fits the description of Pancreatic tissue (The pancreas, 2012).
On ME picture #4 a cell containing a large nucleus with dominating amount of weatherization, extensive rough endoplasmic reticulum with a few organelles were observed. Large amount of weatherization, non transcribed eukaryotic chromatin, result in only limited need for transcription by the high degree of specialization and the short life-span. Furthermore, the extensive rough RE implies the production of good deal of protein. Plasma cells are the B cells that float in the blood stream and synthesize and secrete antibodies . Thus we concluded that the ME picture is a plasma cell (personal communication with roof.
Datelined, 12. 04. 2013). On ME picture #5 shows glycogen, microbes, gall capillaries and a sinusoid.. Glycogen, glucose storage polysaccharide, is energy storage form for the cells. Microbes (lissome and peroxides) are responsible for the storage of acid hydrolysis enzymes that break down waste components and degrade of hydrogen peroxide respectively executing bio-detoxification. The presence of gall capillaries indicate a connection to the gall bladder while a sinusoid with microvolt indicates absorption characteristics as they are essential for the metabolic and detoxification functions.