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首页> 外文会议>Optical Investigations of Cells In Vitro and In Vivo >Autofocus for phase-contrast microscopy: investigation of causes of nonunimodality
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Autofocus for phase-contrast microscopy: investigation of causes of nonunimodality

机译：相聚焦显微镜自动聚焦：非单峰性原因的调查

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Abstract: Autofocus functions based on the measurement of the high spatial frequency content of the image have proven most precise and robust for microscopy. In some situations, however, the through-focus response of the highpass autofocus functions exhibit local maxima, or side peaks. These violations of unimodality can produce spurious autofocus results. Through-focus phase-contrast images of a stage micrometer and their spectra were studied in order to identify the conditions under which these side peaks are generated. The through-focus spectra suggest the side peaks occur when the bandpass of the filter includes the frequencies that undergo contrast reversals. These contrast reversals occur as a fundamental characteristic imposed by the optical system. Contrast reversals, however, are not seen in the frequency range near the optical cutoff. Thus, it was possible to design narrow bandpass filters that isolated only these frequencies and produced unimodal autofocus functions. Based on unimodality, improved sharpness, and consideration of the conditions present in scanning applications, sharply defined highpass filters should prove best for general microscopy autofocus. !7
机译：摘要：基于对图像的高空间频率内容的测量，自动对焦功能已被证明对显微镜最精确，最可靠。但是，在某些情况下，高通自动对焦功能的全焦点响应会表现出局部最大值或侧峰。这些违反单峰性的行为会产生虚假的自动对焦结果。为了确定产生这些副峰的条件，研究了载物台测微计的全焦点相衬图像及其光谱。通过焦点的频谱表明，当滤波器的带通包括经历对比度反转的频率时，会出现侧峰。这些对比度反转是由光学系统施加的基本特征。但是，在光截止附近的频率范围内看不到对比度反转。因此，有可能设计出窄带通滤波器，该滤波器仅隔离这些频率并产生单峰自动聚焦功能。基于单峰性，提高的清晰度以及对扫描应用程序中存在的条件的考虑，清晰定义的高通滤镜应该证明是常规显微镜自动聚焦的最佳选择。 ！7

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