期刊
APPLICATIONS IN PLANT SCIENCES
卷 3, 期 4, 页码 -出版社
BOTANICAL SOC AMER INC
DOI: 10.3732/apps.1400112
关键词
confocal microscopy; fluorescence; morphology; optical sectioning; petal cells
资金
- National Science Foundation Doctoral Dissertation Improvement Grant [DEB-1406650]
- Division Of Environmental Biology
- Direct For Biological Sciences [1406650] Funding Source: National Science Foundation
Premise of the study: Visualizing flower epidermal cells is often desirable for investigating the interaction between flowers and their pollinators, in addition to the broader range of ecological interactions in which flowers are involved. We developed a protocol for visualizing petal epidermal cells without the limitations of the commonly used method of scanning electron microscopy (SEM). Methods: Flower material was collected and fixed in glutaraldehyde, followed by dehydration in an ethanol series. Flowers were dissected to collect petals, and subjected to a Histo-Clear series to remove the cuticle. Material was then stained with aniline blue, mounted on microscope slides, and imaged using a compound fluorescence microscope to obtain optical sections that were reconstructed into a 3D image. Results: This optical sectioning method yielded high-quality images of the petal epidermal cells with virtually no damage to cells. Flowers were processed in larger batches than are possible using common SEM methods. Also, flower size was not a limiting factor as often observed in SEM studies. Flowers up to 5 cm in length were processed and mounted for visualization. Conclusions: This method requires no special equipment for sample preparation prior to imaging and should be seen as an alternative method to SEM.
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