Lab 2 - Techniques of Microscopy

Lab 2 - Techniques of Microscopy - Anil Kanungo BIO 206L...

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8/26/2008 Anil Kanungo BIO 206L Thursday 6-10pm Lab #2: Techniques of Microscopy Introduction: This lab explored the basics of using a light microscope and examining specimen. Students learned to set Köhler, dye specimen, and use digital photography to record specimen. Techniques of microscopy were taught during this lab session. Materials: Olympus Research Microscope Razor Blade 3 Slides with Cover Slips Bottle of 1N HCl Spot Plate Bottle of Aceto-Orcein Solution Pencil (with eraser) CCD Camera and Computer Bottle of PBS (Phosphate Buffered Saline) Forceps Toothpick Bottle of H 2 O Bottle of Methylene Blue Elodea leaf Slime mold plasmodium Kalanchoe leaf Paramecium Procedures: The lab this week involved extensive training focusing and adjusting the Olympus Microscope. In particular, students learned: the effects of lamp intensity, how to move objects on the stage, about the functions of the field and aperture diaphragms, to set Köhler, and to stain specimen. Students cut onion root tips, stained them accordingly, and placed them on slides to study them under a microscope. The TA demonstrated various other specimen. A CCD camera was used for digital imaging and an instructional video was shown preceding the laboratory exercises. Results and Analysis: Exercise 1: Instructional Video Watched video in which exercises 2 and 3 were illustrated. Exercise 2: Focusing the Microscope This exercised involved basic focusing procedures and setting the microscope into focus using a prepared slide containing the following: Image shown at 10x Page 1 of 5
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8/26/2008 Anil Kanungo BIO 206L Thursday 6-10pm Exercise 3: Setting Köhler Köhler was set by adjusting the field diaphragm and the condenser. Exercise 4: Computing Resolution Resolution was computed using the formula d = 0.612λ / NA and recorded on the Reticle Calibration handout. Exercise 5: Onion Root Tip Observed Through Stained Preparation About 2mm of a live onion root tip was cut and placed in a porcelain spot plate. 3 drops were added to the tip and allowed to sit for ~3 minutes in order to help break down cell walls. Using forceps, the root tip was then removed from its current depression and placed into another depression on the spot plate so 3 drops of the aceto-orcein solution could be added. The tip soaked in the solution for another ~3 minutes. The root tip was then placed on the slide and a drop of aceto-orcein solution was placed on the tip to wet mount it. A cover slip wash squashed
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