Decher J, Bachmann P
Histochemistry. 1979 Sep;63(2):221-6. doi: 10.1007/BF00644544.
The amount of light energy a particle absorbs does not depend upon correct focus. The change in the path of light rays brought about by defocusing causes absorbing areas to be registered as areas of higher transmittance than when in focus. Already wellknown in photometry, this effect is put to use by the "blind focus" method at the television texture analysis system (TAS, Leitz). Some chromophores within the object to be measured are compared to a preset value of transmittance, for example T = 0.40. Only the area representing the structures as dense or denser than the preset density are registered. If the structures are out of focus the size of the registered area is too low, since by defocusing, structures to be measured become pale and diffuse, the correct focus corresponds to the largest area to a preset value of transmittance.
粒子吸收的光能数量并不取决于正确对焦。散焦引起的光线路径变化会使吸收区域被记录为比聚焦时具有更高透光率的区域。这种效应在光度学中早已为人所知,电视纹理分析系统(TAS,徕卡)的“盲对焦”方法就利用了这一效应。将被测物体内的一些发色团与预设的透光率值(例如T = 0.40)进行比较。只有代表结构密度等于或高于预设密度的区域才会被记录下来。如果结构失焦,记录区域的尺寸会过小,因为通过散焦,被测结构会变得暗淡且模糊,正确对焦对应于达到预设透光率值的最大区域。