运用图像处理软件将同视野两种单标记免疫荧光图像合成为双标记图像的方法研究

探讨以计算机图像处理技术将普通荧光显微镜采集的同视野单标记免疫荧光图像合成为双标记图像的方法。应用AdobePhotoshop7.0和ImageJ两个图像处理软件,将普通荧光显微镜采集的同视野单标记荧光图像叠加处理成双标记荧光图像,并同激光扫描共聚焦显微镜获得的同一双标记免疫荧光图像进行比较。普通荧光显微镜采集的单标记荧光图像在分辨率及清晰度上逊于激光扫描共聚焦显微镜采集的单标记图像,但显示的脊髓室管膜细胞形态更为完整,细胞突起较长且更连续。因此,在没有激光扫描共聚焦显微镜时,可用AdobePhotoshop7.0或ImageJ图像处理软件将普通荧光显微镜采集的同视野两种单标记免疫荧光图像合成为保持原有特点、简便而清晰的双标记荧光图像。

METHODOLOGICAL STUDY OF SYNTHESIZING DOUBLE-STAINED IMMUNOFLUORESCENCE IMAGE FROM TWO SINGLE-STAINED IMAGES BY USING IMAGE-PROCESSING SOFTWARES

The present study investigated the methodology of synthesizing two single-staining immunofluorescence images into one overlapped double-stained image by using computerized image-processing techniques. With Adobe Photoshop 7.0 and Image J image-processing softwares, two single-stained immunofluorescence images taken with a common fluorescence microscope in the same spinal cord area were synthesized into one overlapped double-stained image and such a synthesized image was compared with the same double-stained image processed with a laser scanning confocal microscope. The single-stained fluorescence images collected with a common fluorescence microscope showed a better shape integrity of the spinal ependymal cells and longer uninterrupted processes, although the capacity and defining powers of the images are not as predominant as those of the same images processed with the confocal microscopy. Thus, in the absence of a confocal microscope, co-field double-stained immunofluorescence images can be obtained from single-stained images collected with a common fluorescence microscope using Adobe Photoshop 7.0 or Image J image-processing techniques.