Amano E
Environ Health Perspect. 1981 Jan;37:165-8. doi: 10.1289/ehp.8137165.
Pollen grains are haploid gametes of uniform shape and size, and can be obtained in large quantity. If appropriate traits are used, they can be an excellent material for investigation of rare but important biological events like intracistronic recombinations or mutations induced by very low level of mutagens. This advantage will be further improved, if the laborious counting and examination can be made automatically. For automation of pollen analysis, techniques of flow analysis and image analysis would be applicable. Flow analysis with a optical detector was tested using maize pollen. Pollen grains were transported by gentle suction through a glass capillary which was placed under a microscope. Interruptions of the light path by pollen grains were detected by a silicon photocell after optical magnification and converted into electric pulses. The frequency distribution of pulse height was examined by a multichannel pulse height analyzer. 10(6) pollen grains would be counted and classified within about 30 min for a pollen suspension dilute enough for separation of each pulse. The flow system tested seems promising for detection of Wx mutant pollen in a wx pollen population after iodine staining if illumination of sample particles is improved.
花粉粒是形状和大小均一的单倍体配子,且能够大量获取。如果使用合适的性状,它们可以成为研究罕见但重要的生物学事件(如顺反子内重组或极低水平诱变剂诱导的突变)的优良材料。如果能够自动进行繁琐的计数和检测,这一优势将得到进一步提升。对于花粉分析的自动化而言,流动分析和图像分析技术都是适用的。使用玉米花粉对配备光学检测器的流动分析进行了测试。花粉粒通过轻柔的抽吸作用经置于显微镜下的玻璃毛细管输送。花粉粒对光路的阻断在光学放大后由硅光电管检测到,并转换为电脉冲。脉冲高度的频率分布由多通道脉冲高度分析仪进行检测。对于稀释程度足以使每个脉冲分离的花粉悬浮液,在约30分钟内可对10⁶个花粉粒进行计数和分类。如果能够改善样品颗粒的照明条件,所测试的流动系统在碘染色后检测wx花粉群体中的Wx突变体花粉方面似乎很有前景。