Any scientific study begins with the compilation of data, either from primary or secondary sources. Some of these experiments, however, require a systematic and thorough procedure carried out in laboratories to establish the theory being studied or to observe the mechanisms of a given microorganism (Valdes & Yin, 2016). This paper aims to critically carry out a lab study on the basic, Gram, and endospore stains on bacterial cells on the same basis. In addition, this paper will detail the different methods and resources needed in each process, explain the findings, and draw a conclusion based on the results. Besides, it will also explain why staining is crucial in the process of identifying bacteria, the differences between differential and simple stains and how staining is used on a regular basis.
Staining is important in the course of identifying bacteria since it is an auxiliary method employed in microscopy to help improve contrast in the microscopic images. Again, it is important in the sense that it contributes to highlight structures of microorganisms’ tissues for better viewing usually with the aid of different microscopes. Staining is also crucial because it helps in determining which bacteria are positive and negative as in the case of Gram staining. On the other hand, there are big differences between differential and simple stains (Savov et al., 2014). For instances, differential staining needs the use of three chemicals while simple stains require stains of different colors. Again, simple staining is different from differential staining in that stain in simple staining method stains all cells equally while stains in differential staining specific cells do not show especially when it is suspected that there is more than one type of bacteria present in the cell. On a regular basis, staining is employed in microscopes to check bacteria images. They are frequently used in medicine, chemistry and biology experiments to look at the organic tissues of various bacteria.
In this research different methods and materials were used for the simple, Gram and Endospores stains. In the simple stains, materials such as Methylene Blue dye, heat source, slides, dropper, water, a microscope and a round shape cocci were used (Rénes, Jakab, Nehme, & Nehme, 2015). The procedure for determining the structures of this micro- organism include; coming up with two cycle lines at the bottom of the slide and naming the frosted end TOP. This is followed by equally smearing E. coli broth culture on one of the drawn cycles on top of the slide. On the remaining cycle, a small amount of water is removed from it while a loop full of water is added. The slide is left to dry for some minutes then heated to fix the plate. The heated attached slide is subjected to Methylene Blue dye which is added by a dropper while the excess dye is eliminated with the help of water. The slide is then observed under a microscope.
On the other hand, in Gram stain, materials such as the source of heat, bacteria, slide microscope water and staining jar were used. Its procedure involves heating smears of the negative and positive Grams by passing them over a flame thrice. They are then put in a staining jar with crystal violet for one minute then washed to get rid of excess stains while excess water is removed tilting the slide. This is followed by putting the slide in a staining jar having mordant for at least one minute then rinsed and decolorized for 12 minutes. Again the slide is rinsed and placed on a piece of bibulous paper and then observed under a microscope. Besides that, in the endospore staining, materials such as bacteria, bibulous paper, Malachite Green, and microscope are used (Greilhuber & Doležel, 2009). The procedure in this stain involves, smearing the bacteria and covering it with a bibulous paper. This is followed by steaming the slide with Malachite Green for 10 minutes in a steamer. The slides are then removed so as to cool and then rinsed with water and counterstaining arrived Safranin for one minute. Lastly, the slide is rinsed blotted and observed with a microscope.
After all these procedures nothing was visible for the observation in any of the slides under the microscope. The possible causes of this problem may have been, either the decolorized was left for a long period thus affecting the cell of the organisms to be observed, or the cell was heated for a long time (Arango Aramburo, Castañeda Acevedo, & Olaya Morales, 2012). Another factor can be that the cells were left to dry for a long time when placed on the Malachite Green or it was blotted with paper towel hence tampering with the cell making it hard to observe.
Based on the result from the lab, it is true that the experiment did not follow the procedure the thus making it hard to find the correct results. I have learned that when carrying such research, it is crucial to observe the temperatures and other procedures so as not to find a wrong observation. Again, I learned that it is advantageous to use simple staining than differential staining since simple is quick and more reliable as compared to the differential. Also, organisms such as spirilla do not stain well while Bacillious species give a lot of endospores. However, stains affect endospores in that it makes its microorganisms less resistant hence bringing easiness in observing their structures.
Arango Aramburo, S., Castañeda Acevedo, J., & Olaya Morales, Y. (2012). Laboratory experiments in the system dynamics field. System Dynamics Review, 28(1), 94-106. http://dx.doi.org/10.1002/sdr.472
Greilhuber, J. & Doležel, J. (2009). 2C or not 2C: a closer look at cell nuclei and their DNA content. Chromosoma, 118(3), 391-400. http://dx.doi.org/10.1007/s00412-009-0205-9
Rénes, M., Jakab, A., Nehme, K., & Nehme, S. (2015). Laboratory experiments of point fixed glasses. Epitoanyag – Journal Of Silicate Based And Composite Materials, 67(2), 62-65. http://dx.doi.org/10.14382/epitoanyag-jsbcm.2015.10
Savov, E., Trifonova, A., Todorova, I., Gergova, I., Borisova, M., & Ananieva, M. et al. (2014). ASSESMENT OF THE RESISTANCE OF CLINICAL ISOLATES PSEUDOMONAS AERUGINOSA TO QUINOLONONES. Trakia Journal Of Science, 221-226. http://dx.doi.org/10.15547//tjs.2014.03.001
Valdes, R. & Yin, D. (2016). Fundamentals of Pharmacogenetics in Personalized, Precision Medicine. Clinics In Laboratory Medicine, 36(3), 447-459. http://dx.doi.org/10.1016/j.cll.2016.05.006