光学显微与电子成像 1.ppt

* Adjusting Microscope for Koeher Illumination 2. Lamp fills the front aperture of the condenser  Purpose: to achieve Koeher Illumination Tool: eyepiece telescope or Bertrand lens condenser objective diaphragms lamp power * Adjusting Microscope for Koeher Illumination 3. Focus the specimen  Purpose: to achieve Koeher Illumination Tool: eyepiece telescope or Bertrand lens objective focus dials stage controls NOTED: avoid objective plunge into specimen slide * Adjusting Microscope for Koeher Illumination 4. Focus and center the condenser  Purpose: reduce scattered light and improve image contrast condenser field diaphragm lamp power * Adjusting Microscope for Koeher Illumination 5. Adjust the condenser diaphragm  Purpose: to obtain the best resolution and contrast Tool: eyepiece telescope or Bertrand lens condenser diaphragm NOTED:~3/4 of the maximum diameter of the aperture * Adjusting Microscope for Koeher Illumination 6. Adjust the brightness  Purpose: to obtain optimal image brightness voltage neutral density filters NOTED: aperture diaphragm should never be closed downs as a way to reduce light intensity * * * * * * * Detectors for Optical Microscopy Film photomicrography Video imaging (Close Circuit TV) Digital imaging with a charge coupled device (CCD) Scanning microscopy Photomultiplier tube (PMT)-光电倍增管 Avalanche photodiode (APD)-雪崩光电二极管 * * What is Contrast（对比度）? Differences in light intensity, phase, polarization, color create image contrast, which allows individual features and details of the specimen to become visible. Contrast is defined as the difference in light intensity between the object and the adjacent background relative to the overall background intensity. Percent Contrast = [(I(s) - I(b))] / I(b) x 100% * Contrast Mechanisms（都有哪些对比度） Bright field (low contrast) 明场 Phase contrast 相位衬度 Differential interference contrast (DIC) 微分干涉 Dark field contrast 暗场 Polarization micr