Blomquist E, Arro E, Brunk U, Collins V P, Fredriksson B A
Acta Pathol Microbiol Scand A. 1980 Sep;88(5):327-37. doi: 10.1111/j.1699-0463.1980.tb02503.x.
A method is described which allows a comparison in the transmission electron microscope (TEM) of cells with different remaining proliferative capacity from one and the same culture. The method takes advantage of a mini-cloning technique employing hapatotactic palladium islands in combination with micro-dissection and preparation for TEM of islands carrying various numbers of cells after 10 days in culture, when all miniclones have become density dependent growth inhibited. By means of this technique non-dividers were compared with miniclones of dividers composed of five to eight cells originating from single cells. Moreover, large, immotile cells without peripheral ruffling activity, known to be non-dividers, were compared with small, ruffling cells, known to be dividers, in the reflection-interference mode in sparse cultures of living cells, and in the TEM mode as whole cell preparations after critical point drying of cells cultured on formvar-coated, gold EM-grids. Non-dividers proved to contain a moderate number of residual bodies, well developed Golgi areas, and often branched or circular mitochondria; they were thinly spread over the substratum with many focal points of contact, and large areas of close apposition between cell and substratum.
本文描述了一种方法，该方法可在透射电子显微镜（TEM）下对来自同一培养物中具有不同剩余增殖能力的细胞进行比较。该方法利用了一种微型克隆技术，该技术采用趋肝性钯岛结合显微切割，并在培养10天后对携带不同数量细胞的岛进行TEM制备，此时所有微型克隆均已受到密度依赖性生长抑制。通过该技术，将不分裂细胞与由单细胞产生的五到八个细胞组成的分裂细胞微型克隆进行比较。此外，在活细胞稀疏培养的反射干涉模式下，以及在涂有福尔马膜的金EM网格上培养的细胞经过临界点干燥后作为全细胞制备物的TEM模式下，将已知为不分裂细胞的无外周褶皱活性的大的、不活动细胞与已知为分裂细胞的小的、有褶皱细胞进行比较。结果表明，不分裂细胞含有适量的残余小体、发育良好的高尔基体区域，且线粒体常呈分支状或圆形；它们稀疏地分布在基质上，有许多接触焦点，细胞与基质之间有大面积的紧密贴附。
