Light Microscope to Determine Scale of Object

Light Microscope to Determine Scale of ObjectLight MicroscopeSyed IbrahimIntroductionThe development of the microscope has been vital to much scientific advancement in biology (Kriss Kriss 1998). Microscopes allow humans to see objects that would otherwise be unseen by the stark naked eye. The easygoing microscope uses a series of third genus Lenses to magnify an object. The condenser lens align and focus the light from the illumination source through the stage, onto the specimen. (Murphy, 2001) After passing through the specimen, the light goes to the objective lens which collect diffracted light and magnify the image of the specimen, typically 4X, 10X, 40X, or 100X (Murphy, 2001). The light finally reaches the ocular lens. The ocular lens also focus and magnify the image, besides this is typically 10X or 15X (Murphy, 2001). After passing through the ocular lens, the light reaches the observers eyes.Microscopes do not just magnify the image of an object, that also increase it s resolution (Heidcamp et al., 2014). Magnification is the increase in the dimensions of an image, while resolution is the ability to distinguish two components of the image (Alberts et al., 2008). In other words, the gush is the size of the image while the resolution is the clarity or quality of the image (Heidcamp et al., 2014). There is no limit of magnification because the size of an image nominate be increased indefinitely, but there is a limit of resolution because of the properties of light (Alberts et al., 2008). Due to diffraction, the limit of resolution for light microscopes is close to half the wavelength of light divided by the numerical aperture. (Hell, 2007). The numerical aperture is a measure of the number of light rays collected by the objective lens of a microscope, and it is dependent on the refractive advocate and the sine of half of the cone angle (Heidcamp et al., 2014). These can be combined to give the following equation (Heidcamp et al., 2014)Where = wav elength of light = refractive index = half of the cone angleBased on the above equation, decreasing the wavelength of light, increasing the refractive index, or increasing the cone angle will subside the limit of resolution, thus increasing the resolution of an image. The smallest limit of resolution of a light microscope is 0.2m (Alberts et al., 2008).Microscopes can be used to examine microorganisms. In this testing ground Spirogyra, Paramecium and Saccharomyces cerevisiae were examined. Spirogyra are filamentous algae that are typically 10m-100m wide and their filaments may be a few centimeters hanker (Parmentier, 1999). Spirogyra are often launch in freshwater are distinguishable by their spiral chloroplasts (Fathima et al., 2007). Paramecium are unicellular protists with cilia that are typically found in aquatic habitats and are usually 100m-3500m (Morgan, 1999 Wichterman, 1986). Saccharomyces cerevisiae (yeasts) are unicellular fungi that are typically 3m-6m in size (Schne iter, 2004). Since the naked eyes limit of resolution is 100m, these organisms are too small to be observed by the human eye solo (Heidcamp et al., 2014). Light microscopy was used to increase magnification and resolution so that the individual organism as intimately as their internal structures may be distinctly observed.The purpose of this lab was to use a bright field microscope to determine the scale of each objective, to examine Spirogyra, Paramecium, wild-type yeasts and fab1 mutant yeasts under a microscope, as well as to learn the essentials of micropipetting.ResultsPart A Lab 1 Report SheetsPlease refer to attached sheets.Part B Answers to Assigned QuestionsWhen the dimensions for the letter e using 4X, 10X or the naked eye were compared in good example 1.2, they were all approximately the same, as seen below. Using the light microscope gave more precise dimensions as compared to the naked eye. When comparing the different magnifications of the light microscope, they ha d percentage differences of 4% and 8% in the length and width respectively. Overall, it makes sense that all three measurements gave roughly the same dimensions as they were all measuring the same specimen.Dimensions of the letter eNaked Eye Light Microscope (4X) Light Microscope (10X) Percentage difference between 4X and 10XBased on the observations from action 1.3, it was apparent that Spirogyra have cell walls while Paramecium do not. As well, Paramecium have cilia while Spirogyra do not.After pipetting as required for Exercise 1.4, a minute amount of water remained in the Eppendorf tube, and there was no air gap in the tip of the pipette. This means that slightly more than 50L of water was pipetted into the Eppendorf tube. For this reason we practiced again, and this time no liquid remained. For future labs, we must ensure that we are extra attentive to ensure we pipette the correct amount of liquid.During Exercise 1.5, it was observed that fab1 mutant yeasts appeared to have a thicker cell membrane than the wild-type yeasts. This thicker cell membrane may have been an enlarged vacuole at heart the cell that was pressing up against the cell membrane.Part C ResearchThere are many types of light microscopes, including bright-field microscopes, dark-field microscopes and phase-contrast microscopes (Alberts et al., 2008). Phase contrast microscopes rely on the phase-shifting of light as it passes through parts of the specimen of different relative thickness and density (Zernike, 1942).Search Engine Web of ScienceSearch legal injury phase contrast microscopic filtered by date from 1900 to 1950Reference Zernike, F. (1942). Phase contrast, a newly method for the microscopic observation of transparent objects.Physica,9(7), 686-698.After researching, a microscope was found with the following specifications and price (Cole-Parmer, 2014)Microscope Phase Contrast Microscope with Digital Camera (3 megapixels), Binocular, 115 VAC, 60 HzDistributer Cole-ParmerModel Number RK-48925-04Approximate equipment casualty $2,932.46CND/EACHSearch Engine GoogleSearch Terms Phase Contrast Microscope with Digital CameraReference Cole-Parmer. (2014). Phase Contrast Microscope with Digital Camera.Cole-Parmer. Retrieved September 15, 2014, from http//www.coleparmer.ca/ growth/Phase_Contrast_Microscope_with_Digital_Camera_Binocular_115_VAC_60_Hz/RK-48925-04ReferencesAlberts, B., Johnson, A., Lewis, J., Raff, M., Roberts, K., Walter, P. (2008).Molecular Biology of the Cell(5th ed.). New York Garland Science.Cole-Parmer. (2014). Phase Contrast Microscope with Digital Camera.Cole-Parmer. Retrieved September 15, 2014, from http//www.coleparmer.ca/Product/Phase_Contrast_Microscope_with_Digital_Camera_Binocular_115_VAC_60_Hz/RK-48925-04Fathima, M., Shantha, N., Rajagovindan, N. (2007).Botany(Revised ed.). Chennai Tamil Nadu Textbook Corporation.Heidcamp, W., Antonescu, C., Botelho, R., Victorio-Walz, L. (2014).Laboratory Manual Cell Biology BLG311(Fall 2014 ed. ). Toronto Ryerson University.Hell, S. W. (2007). Far-Field Optical Nanoscopy.Science,316(5828), 1153-1158.Kriss, T. C., Kriss, V. M. (1998). fib of the Operating Microscope From Magnifying Glass to Microneurosurgery. Neurosurgery,42(4), 899-907.Morgan, M. (1999). Paramecium. Microscopy-UK. Retrieved September 15, 2014, from http//www.microscopy-uk.org.uk/index.html?http//www.microscopy-uk.org.uk/ponddip/paramecium.htmlMurphy, D. B. (2001).Fundamentals of light microscopy and electronic imaging. New York Wiley-Liss.Parmentier, J. (1999). Spirogyra. Microscopy-UK. Retrieved September 15, 2014, from http//www.microscopy-uk.org.uk/index.html?http//www.microscopy-uk.org.uk/ponddip/spirogyra.htmlSchneiter, R. (2004).Genetics, Molecular and Cell Biology of Yeast. Fribourg University of Fribourg Switzerland.Wichterman, R. (1986).The Biology of Paramecium(2nd ed.). New York Plenum Press.Zernike, F. (1942). Phase contrast, a new method for the microscopic observation of transparent object s.Physica,9(7), 686-698.