Clear visualization of the products of nonradioactive in situ hybridization in plant tissue by simple dark-field microscopy
✍ Scribed by Sachihiro Matsunaga; Shigeyuki Kawano; Tetsuya Higashiyama; Noriko Inada; Tsuneyoshi Kuroiwa
- Publisher
- Elsevier Science
- Year
- 1997
- Tongue
- English
- Weight
- 287 KB
- Volume
- 28
- Category
- Article
- ISSN
- 0968-4328
No coin nor oath required. For personal study only.
✦ Synopsis
Nonradioactive in situ hybridization is a powerful technique for determining levels of mRNA that have accumulated in tissues, and bright-field microscopy has been used extensively to detect signals generated by nonradioactive in situ hybridization. We converted an inverted microscope to a dark-field microscope by equipping it with a handmade ring-slit. When the in situ hybridization pattern in a plant tissue was examined by this simple method of dark-field microscopy, the hybridization signals appeared clearly bluish-purple within the silvery white cell walls of plant tissues. This method eliminates the necessity for counterstaining because the silvery white cell walls clearly reveal the shape of the plant tissues. Furthermore, color reproduction of processed images obtained by simple dark-field microscopy clearly reveals the sites of expression of mRNA.