Fixation, embedding, and sectioning of plant tissues.
CSH Protocols
|March 2, 2011
Summary
This protocol details preparing plant tissues for sectioning using Paraplast embedding. This method is crucial for subsequent immunohistochemistry and in situ hybridization analyses.
Area of Science:
- Plant Biology
- Molecular Biology
- Histology
Background:
- Accurate tissue preparation is essential for reliable molecular analyses in plant science.
- Immunohistochemistry and in situ hybridization require well-preserved tissue morphology and antigen/nucleic acid integrity.
Purpose of the Study:
- To provide a detailed protocol for preparing plant tissues for high-quality sectioning.
- To enable successful downstream applications like immunohistochemistry and in situ hybridization.
Main Methods:
- Plant tissues are fixed to preserve cellular structure.
- Tissues are embedded in Paraplast, a paraffin wax medium, for stable support.
- Embedded tissues are sectioned to create thin slices suitable for microscopy.
Main Results:
- The protocol yields consistently sectionable plant tissues.
- Preserved morphology and molecular targets are suitable for advanced imaging techniques.
Conclusions:
- This Paraplast embedding protocol is effective for plant tissue preparation.
- It serves as a foundational step for detailed molecular and cellular investigations in plants.
Related Concept Videos
01:03Fixation and Sectioning
Two basic types of preparation are used to visualize specimens with a light microscope: wet mounts and fixed specimens.
The simplest type of preparation is the wet mount, in which the specimen is placed in a drop of liquid on the slide. A liquid specimen can be directly deposited on the slide using a dropper. Solid specimens, such as skin scraping, can be placed on the slide before adding a drop of liquid to prepare the wet mount. Sometimes the liquid is simply water, but stains are often added...
The simplest type of preparation is the wet mount, in which the specimen is placed in a drop of liquid on the slide. A liquid specimen can be directly deposited on the slide using a dropper. Solid specimens, such as skin scraping, can be placed on the slide before adding a drop of liquid to prepare the wet mount. Sometimes the liquid is simply water, but stains are often added...
01:20Preparation of Samples for Electron Microscopy
To be visualized by an electron microscope, either transmission or scanning, biological samples need to be fixed (stabilized) so the electron beam does not destroy them and dried thoroughly (desiccated/dehydrated) so the vacuum does not affect them. Fixation needs to be done as quickly as possible because the sample properties will start changing as soon as it is removed from its natural environment. For example, in a tissue sample, the oxygen levels begin decreasing, causing an altered...