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General Mic Study Guide

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Microbiology lab study guide Exam 1
Know step by step instructions on use of microscope to view a specimen (image remains in focus even when changing objective lens – parfocal) * Bright field- Dark image against bright background. * Dark field- Field surrounding the specimen appears black while object is brightly illuminated. * Phase contrast- Specimens appear as various levels of “darks” against a bright background. * Fluorescence- fluorescence dye is used which emits fluorescence under UV light.. * Total Magnification = Magnification by the objective lens X Magnification by ocular lens. * FOCUS MICROSCOPE PROPERLY by starting with scanning lens and moving up each objective, using scope’s parfocal property

Know step by step instructions for ocular micrometer calibration using the stage micrometer * Calibration = Stage Micrometer /Ocular Micrometer * Stage Micrometer: mm convert to um * Ocular Micrometer: 0 – 100 Ocular Units * EX: 0.2mm X 1,000 um = 200 um * 1mm * Placed stage micrometer superimposed by the ocular micrometer. * First left line of the ocular micrometer aligned with one of the marks on the stage micrometer * EX 25 of ocular micrometer on 8 major lines of the stage micrometer are perfectly aligned= 800 µm ÷ 25 ocular units = 32 um/OU * Record more than one measurement * Calibrate each magnification *
Know the different parts of the microscope – stage, course & fine adjustment knobs, etc

Know the definitions of parfocal, resolution, numerical aperture etc. * Parfocal -Image remains in focus when objectives are changed requiring only minor adjustments with the fine focus knob * Resolution The ability of the microscope to separate close details. The light microscope cannot resolve objects smaller than 0.2 µm. Specimen

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