Laboratory for Applied Microscopy Research, Marienburgstr. 23, D-56859, Bullay, Germany.
Microsc Res Tech. 2013 Oct;76(10):1050-6. doi: 10.1002/jemt.22265. Epub 2013 Jul 30.
Using the optical methods described, phase specimens can be observed with a modified light microscope in enhanced clarity, purified from typical artifacts which are apparent in standard phase contrast illumination. In particular, haloing and shade-off are absent, lateral and vertical resolution are maximized and the image quality remains constant even in problematic preparations which cannot be well examined in normal phase contrast, such as specimens beyond a critical thickness or covered by obliquely situated cover slips. The background brightness and thus the range of contrast can be continuously modulated and specimens can be illuminated in concentric-peripheral, axial or paraxial light. Additional contrast effects can be achieved by spectral color separation. Normal glass or mirror lenses can be used; they do not need to be fitted with a phase plate or a phase ring. The methods described should be of general interest for all disciplines using phase microscopy.
利用所描述的光学方法,可以在改良的光显微镜下观察具有增强清晰度的位相标本,使其免受标准相衬照明中明显的典型伪像的影响。特别是,不存在晕光和消光现象,横向和纵向分辨率最大化,并且即使在难以用正常相衬检查的有问题的制剂中,图像质量也保持不变,例如超过临界厚度的标本或被倾斜盖玻片覆盖的标本。背景亮度因此对比度范围可以连续调节,可以对标本进行同心-外围、轴向或傍轴照明。通过光谱颜色分离可以实现额外的对比效果。可以使用普通玻璃或反射镜透镜;它们不需要配备相板或相环。对于使用相衬显微镜的所有学科来说,所描述的方法都应该具有普遍的意义。