Tang C, Biemond I, Lamers C B
University Hospital, Leiden, The Netherlands.
Biotechniques. 1995 May;18(5):886-9.
The present study evaluated a new imaging technique that demonstrated the application of storage phosphor autoradiography in the localization and quantification of cholecystokinin receptors in rat brains and compared the results with film autoradiography. Cryostat sections were incubated with [125I]Bolton-Hunter-labeled sulfated cholecystokinin octapeptide followed by exposure to a storage phosphor-imaging screen and suitable autoradiography film. To obtain satisfactory images, it took 6 days with film autoradiography vs. 15 hours with the storage phosphor technique. Both film and storage phosphor autoradiograms showed the same cholecystokinin receptor distribution in brain sections; however, the film imaged more details. To reach the lowest possible response ratio between low and high receptor density regions in rat brains, storage phosphor autoradiography was about 240-fold faster than film. In addition, the new technique presented a considerably larger exposure time range for maintaining that ratio. The binding per area showed a linear relationship with the thickness of sections between 5 and 14 microns. In the linear response range, the quantitative results of both methods are comparable. In conclusion, storage phosphor autoradiography is a faster technique for localizing and quantifying peptide receptors in tissue sections but slightly compromised in resolution when compared with film autoradiography.
本研究评估了一种新的成像技术,该技术展示了存储磷光体放射自显影术在大鼠脑内胆囊收缩素受体定位和定量中的应用,并将结果与胶片放射自显影术进行了比较。将冷冻切片与[125I]博尔顿-亨特标记的硫酸化胆囊收缩素八肽孵育,然后暴露于存储磷光体成像屏和合适的放射自显影片上。为获得满意的图像,胶片放射自显影需要6天,而存储磷光体技术需要15小时。胶片和存储磷光体放射自显影片均显示脑切片中胆囊收缩素受体分布相同;然而,胶片成像的细节更多。为使大鼠脑内低受体密度区和高受体密度区之间的响应比达到尽可能低的水平,存储磷光体放射自显影术比胶片快约240倍。此外,新技术在维持该比例方面呈现出大得多的曝光时间范围。每单位面积的结合与5至14微米厚的切片厚度呈线性关系。在该线性响应范围内,两种方法的定量结果具有可比性。总之,存储磷光体放射自显影术是一种用于组织切片中肽受体定位和定量的更快技术,但与胶片放射自显影术相比,分辨率略有下降。