Giles J J, Kavanagh H J, Bannigan J G
Department of Human Anatomy and Physiology, University College, Dublin 2. Ireland.
J Microsc. 1995 Sep;179(Pt 3):282-5. doi: 10.1111/j.1365-2818.1995.tb03643.x.
The chick embryo shell-less culture technique allows continuous observation and access to the developing vessels of the area vasculosa (AV); hence, its value as an angiogenesis assay system. The main drawback of the method is that adequately contrasted photographs cannot be achieved without injection of a contrast medium which kills the embryo making further observation of the same specimen impossible. Furthermore, injection prior to 72 h incubation has a high failure rate. In an attempt to overcome these problems, we explored the possibility of using a high-contrast photographic process. Embryos were explanted into shell-less culture after 48 h incubation and photographed through a stereo microscope. After trials with different films we found that Kodalith ortho Type 3 (Kodak) and Technical Pan film (Kodak) produced images of the vasculature which were identical in contrast and detail to India-ink-injected specimens photographed with conventional film. In addition, excellent images could be recorded as early as 48 h of incubation.
鸡胚无壳培养技术能够对血管区(AV)正在发育的血管进行连续观察并直接接触;因此,它作为一种血管生成检测系统具有重要价值。该方法的主要缺点是,在不注射造影剂的情况下无法获得对比度足够的照片,但注射造影剂会杀死胚胎,导致无法对同一标本进行进一步观察。此外,在孵化72小时之前进行注射的失败率很高。为了克服这些问题,我们探索了使用高对比度摄影方法的可能性。在孵化48小时后将胚胎移出进行无壳培养,并通过立体显微镜进行拍照。在对不同胶片进行试验后,我们发现柯达正色3型胶片(柯达公司)和技术全色胶片(柯达公司)所产生的脉管系统图像,其对比度和细节与用传统胶片拍摄的注射印度墨水的标本相同。此外,早在孵化48小时时就能记录到出色的图像。