Excercise 3 analysis - Nabeel Siddiqui nhs 256 Bio 206L...

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Nabeel Siddiqui nhs 256 Bio 206L Wed. 9:00-1:00 (49530) 2/7/2010 Microscopy Techniques Exercise # 3: Analysis 1. When using higher power objectives, we must use the oil immersion technique in order to account for the high refractive index of light. This causes the light coming through the specimen to be lost before entering the objective. The oil helps to counter the refractive index of light. To place immersion oil on a slide, one should focus on the specimen on a lower power objective such as 40x. After the specimen is in focus, one should turn the nose piece of the objectives halfway between the 40x and 100x objectives. Using a dropper, one should place one drop of oil directly onto the specimen on the slide without moving the slide itself. Then one should turn the 100x objective into place and focus the specimen as needed. The x y knobs should not have to be adjusted if the specimen was in focus while using the lower power objective. As long as the field of view has not changed, the specimen should still be in the focal plane because only the magnification has changed. When going back to a lower power objective after using the immersion oil, one should clean the excess oil off the 100x objective using a specific lens cleaner. Also one should clean the excess oil off the slide before turning into a lower power objective. 2. Based on our observations, the red blood cells seem to have no nucleus and appear to be concave in shape. There are not too many detailed intracellular features apparent in the red blood
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cells. Red blood cells seem to outnumber the white blood cells by a ratio of 1000:1 and seem to be much smaller in size compared to the white blood cells. The white blood cells are purple in color compared to the red blood cells. They are much larger and less frequent than the red blood cells. They have a large circular shape and a dark nucleus present in the specimen. Surface Area Volume Red Blood Cells 1.53 micrometers ² 0.18 micrometers ³ White Blood Cells 3.14 micrometers ² 1.57 micrometers ³ Sample Calculations: SA= 4π (3.5)² Volume= 4/3 π (3.5)³ 3. Phase contrast microscopy solves the problem of contrast. Compared to brightfield illumination, phase contrast gives a more detailed and contrasted image of the specimen. The lines and structures of the specimen are more prominent and offer a greater contrast. This is possible because of how the light in phase contrast microscopy behaves. The wavelengths of light are either phased or shifted when the pass through the specimen. When they are phased, a constructive wavelength is formed which allows for a better image in those particular
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This note was uploaded on 01/25/2011 for the course BIO 206L taught by Professor Unknown during the Spring '08 term at University of Texas at Austin.

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Excercise 3 analysis - Nabeel Siddiqui nhs 256 Bio 206L...

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