| Fluorescent microscopic imaging technology is widely used in zoology, neuroscience, cell biology, molecular biology, and other research fields due to its specificity and contrast. The most commonly used detection instruments for fluorescence microscopy imaging are fluorescence microscopy and in vivo imagers, although in vivo imagers have not been extensively reported for plant detection and analysis. This study analyzed the wheat seeds containing red fluorescent markers (DsRed) and green fluorescent markers (GFP) using fluorescence microscopy, in vivo imaging, and PCR comparative observation. The results showed that the fluorescence expression observed with fluorescence microscopy was mainly located at the root tip and young bud growth regions, allowing detailed observation of individual samples. However, due to the limited field of view of the microscope, conducting large-scale analysis was challenging.The in vivo fluorescence imaging instrument could simultaneously observe a large number of plants with high throughput. When observing GFP and RFP, these markers could be clearly distinguished from the wild-type exhibiting high specificity and minimal background noise. Moreover, unlike fluorescence microscopy, the fluorescence intensity of RFP was higher and brighter. After PCR identification and analysis, both devices were able to correctly screen positive plants, and the results were consistent. This study provides theoretical basis and technical support for the identification and screening of transgenic plants. |
