I am looking for people who tried different coatings on glass slides or petri dishes in order to fix plant cells. I have suspension cells that I want to see in a liquid medium, I tried poly-l-Lysine but the result is not satisfiying.
I know differetn coatings exist for animal cell observations, like gelatin, streptavidin and other types that will attach the plsma membrane by adhesion. The presence of a cell wall in plant cells is an additional challenge. Anyone tried to fix those cells with pectins or other substances? Help please!
Plant tissues must be dehydrated for observation in most electron microscopes. Although a number of sample processing techniques have been developed for preserving plant tissues in their original form and structure, none of them are guaranteed artefact-free. The current paper reviews common scanning electron microscopy techniques and the sample preparation methods employed for visualisation of leaves under specific types of electron microscopes. Common artefacts introduced by specific techniques on different leaf types are discussed. Comparative examples are depicted from our lab using similar techniques; the pros and cons for specific techniques are discussed. New promising techniques and microscopes, which can alleviate some of the problems encountered in conventional methods of leaf sample processing and visualisation, are also discussed. It is concluded that the choice of technique for a specific leaf sample is dictated by the surface features that need to be preserved (such as trichomes, epidermal cells or wax microstructure), the resolution to be achieved, availability of the appropriate processing equipment and the technical capabilities of the available electron microscope.
Scanning electron microscopy (SEM) is an ideal technique for examining plant surfaces at high resolution. Plant tissues must be preserved by dehydration for observation in an electron microscope because the coating system and the microscopes operate under high vacuum and most specimens cannot withstand water removal by the vacuum system without distortion1.
In order to examine the native structure of the sample, some microscopes are designed to image frozen hydrated samples and more recently environmental SEM microscopes have been developed which can image the sample in their native-hydrated state. These microscopes are specialised equipments and may not be available in many labs. Hence, sample preparation by dehydration is still an important consideration for observation in conventional microscopes.
For samples that necessitate dehydration, many techniques other than just air-drying have been developed to remove water from the sample, all aiming at minimal distortion of the cell and maximal preservation of the original form and structure. These techniques include freeze-drying, critical point drying, and various types of chemical fixation treatments prior to dehydration of samples. However, acceptable methods offer less than ideal preservation for some plant species and may be inconsistent. The inconsistency is largely due to diversity in tissue types, form, structure and composition of plants. Inconsistencies also arise from variation in individual skills and equipment used across different labs. Hence, new/modified techniques are continually being tested and developed for the preparation of specific plant tissues for visualisation under electron microscopes.
Thank you for you help. I am trying to measure plasticity with AFM and not SEM. The real challenge is how to attach suspension cells without fixing them or alter their viability.
May I suggest a reading that might help................I am not of much help:
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2913783/
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