Roxanne Diane R. Uy MST BIO MICROSCOPE AND MICROMETRY ACTIVITY # 2 I. Introduction A.

Background Microscopes are intended to measure small objects. Size is one of the things needed to be measured using microscope, which can be achieved by using the rulers of a microscope-ocular and stage micrometer. The ocular micrometer (see Figure 1-A), known as an arbitrary ruler, has no assigned unit since the size of a magnified image will change depending on the objective being used. Therefore, it must be calibrated with a stage micrometer (see Figure 1-B), for each of the objective lens, in order for it to have a quantitative value. A stage micrometer is considered to be the true ruler, since it has units (millimeters (mm) or micrometers (m)). When calibrating, one will line up the stage micrometer with the ocular micrometer and count the number of divisions on the ocular micrometer per millimeter or micrometer on the staged micrometer. The number of divisions will change as the magnification changes.

B. Question Q1: What is the relationship of magnification of the objective lens (A) with the calibration constant of the micrometer (B)? C. Objective To determine the importance of calibration in doing accurate measurement To establish the relationship of magnification of the objective lens and the calibration constant of the micrometer. D. Hypothesis H0: There is no significant relationship between A and B H1: There is a significant relationship between A and B II. Materials and Method

A B C Figure 1. A-Ocular micrometer, B- Stage micrometer, C- Ocular micrometer superimposed on a stage micrometer

B C Figure 2. Materials used for the activity. A-Compound microscope, B- Stage micrometer, C- Ocular micrometer

Obtain a stage and ocular micrometer (see Figure 2). Place the stage micrometer on the stage of the microscope. Rotate one of the microscope eyepieces until the lines of the ocular micrometer are parallel with those of the stage micrometer. Match the lines on the left edges of the two micrometers by moving the stage micrometer so that the graduations of the ocular micrometer are superimposed over those of the stage micrometer. Determine the number of ocular micrometer spaces that fall within a given number of stage micrometer spaces. Calculate the distance between each ocular graduation by using the following formula: 1 ocular micrometer space (m) = x spaces on the stage micrometer y spaces on the ocular micrometer Repeat the procedure for the 10X, 40X, and 100X objectives and record results. After acquiring the result for all objective lens, compute the calibration constant using the following formula: Calibration Constant = Stage Micrometer Ocular Micrometer x 0.01mm

Table 2. Result of Pearson Correlation of the Calibration Constant and the Magnification of Objective Lenses
Pearson Product Moment Correlation Statistic Mean Biased Variance Variable X 38.5 1446.75 Variable Y 9.675 91.211875

Biased Standard Deviation 38.0361669993179 9.55049082508329 Covariance Correlation Determination T-Test -362.85 -0.749145117857612 0.561218407609895 -1.5993987470537 0.250854882142388 0.125427441071194 2

III. Results and Discussion Table 1. Average calibration constant per magnification of the objective lens of 13 students Objective Lens LPO HPO (10x) (40x) 10.1 2.5

p-value (2 sided) p-value (1 sided) Degrees of Freedom

Calibration Constant Mean

Scanner (4x) 25.1

IOI (100x) 1.0

Number of Observations 4

IV. Conclusion Based on the results, I therefore reject the null hypothesis and accept the alternative hypothesis-magnification and calibration has a strong relationship. As the magnification of the objective lens increases, the calibration constant decreases, establishing a inverse relationship among the two. OBJECTIVES V. References Agar Scientific (n.d.). Calibration standards and specimens. Retrieved July 8, 2013, from http://www.agarscientific.com/media/import/03_Calibrati on_Standards_pgs_39-84_date_17_06_10_web.pdf City College of San Francisco (n.d.). Quantitative microscopy. Retrieved July 8, 2013, from http://fog.ccsf.cc.ca.us/dvandepo/documents/microscopy_ quant.pdf Damon, A. W. (2011). Micrometer eyepiece. Retrieved July 8, 2013, from http://www.mrdamon.com/experiments/1ib_bio/micrometer.htm DoctorFungus (n.d.). Ocular micrometer. Retrieved July 8, 2013, from http://www.doctorfungus.org/thelabor/sec11.pdf Gardner, D. (2007). Measuring with a microscope. Retrieved July 8, 2013, from http://www.ccmr.cornell.edu/ret/modules/documents/Mea suringwithaMicroscope.pdf

Figure 3. Shows the Magnification of the Objective Lens against the averaged calibration constant Table 1 shows the magnification of the different objective lens, from the lowest (Scanner-4x), to the highest (IOI-100x). Mean of the calibration constant is also shown having a downward trend. To test for the relationship of the two variables, Pearsons relationship was used. Table 2 gives a correlation result of 0.749145117857612 . A linear relationship was established. To add up, a negative coefficients tell us that there is an inverse relationship: when one variable increases, the other one decreases. A result of -.70 or higher indicates a very strong negative relationship between the 2 variables.

Leica DME, & Idaho State University (n.d.). Microscope use. Retrieved July 8, 2013, from http://www.isu.edu/departments/biolab/Training/Microscopes.p df Microtec Microscopes (n.d.). Calibration of microscope eyepiece graticule. Retrieved July 8, 2013, from http://www.tecmicroscopes.co.uk/Downloads/Microtec%20%20graticle%20calibration.pdf Phillips Community College (n.d.). The microscope. Retrieved July 8, 2013, from http://www.biologyjunction.com/Microscope%20Lab.pdf Pyser SGI (2005, September 6). Stage Micrometers. Retrieved July 8, 2013, from http://www.pysersgi.com/images/thumbnails/Graticules/Stage%20Micrometers% 20web.pdf University of Cambridge International Examinations (2006). Teaching AS biology practical skills. Retrieved July 8, 2013, from http://papers.xtremepapers.com/CIE/Cambridge%20Internation al%20A%20and%20AS%20Level/Biology%20(9700)/9700_no s_ps_2.pdf Wessa P., (2012), Pearson Correlation (v1.0.6) in Free Statistics Software (v1.1.23-r7), Office for Research Development and Education, URL http://www.wessa.net/rwasp_correlation.wasp/ Zeiss (1998, January). Standard 25 ICS transmitted light microscope. Retrieved from http://www.zeiss.com/C1256F8500454979/0/093ADF70031823 51C1256F8F00392651/$file/b-40-029standard25_ics.pdf