Hjelstuen M H, Rasch-Halvorsen K, Brekken C, Bruland O, de L Davies C
Department of Physics, The Norwegian Institute of Technology, University of Trondheim, Oslo, Norway.
Acta Oncol. 1996;35(3):273-9. doi: 10.3109/02841869609101641.
Penetration and binding of monoclonal antibody (MAb) in multicell osteosarcoma spheroids have been studied by autoradiography and confocal laser scanning microscopy (CLSM). Optical sectioning of the 3-dimensional spheroids was performed by CLSM. Owing to attenuation of fluorescence intensity, FITC-labelled MAb could not be detected at depths greater than 60 microm within the spheroids. The antibody uptake seen in autoradiographs and CLSM images 60 microm within the spheroids were essentially identical. MAb had reached all parts of the spheroids within 6 h. Quantitative measurements of the fluorescence intensity of FITC-labelled MAb seen in confocal images and measurements of MAb bound per cell using flow cytometry, showed that maximum uptake was reached after 6 h. The possibility to perform both quantitative and qualitative measurements makes CLSM a promising method for studying antibody uptake in thick tissue samples.
通过放射自显影和共聚焦激光扫描显微镜（CLSM）研究了单克隆抗体（MAb）在多细胞骨肉瘤球体中的渗透和结合情况。利用CLSM对三维球体进行光学切片。由于荧光强度的衰减，在球体内深度大于60微米处无法检测到异硫氰酸荧光素（FITC）标记的单克隆抗体。在球体60微米深处的放射自显影片和CLSM图像中观察到的抗体摄取情况基本相同。单克隆抗体在6小时内已到达球体的所有部位。对共聚焦图像中FITC标记的单克隆抗体荧光强度的定量测量以及使用流式细胞术对每个细胞结合的单克隆抗体的测量表明，6小时后达到最大摄取量。能够进行定量和定性测量使得CLSM成为研究厚组织样本中抗体摄取的一种有前景的方法。
