Federica Brandizzi, Ruzin SE. 1999. Plant microtechnique and microscopy . 322 pp. Oxford, New York: Oxford University Press. 32.50 (softback). , Annals of Botany, Volume 86, Issue 3, September 2000, Page 708,
There is a vast literature on plant microtechniques and microscopy proposing a broad collection of protocols with general schedules that have to be optimized depending on the species, the nature of the tissue and the aim of the experiment; therefore each schedule has to be adjusted for every histological experiment to be performed, so that a high quality histological sample could be obtained in the shortest possible time. The availability of ad-hoc schedules for a certain tissue of a determined species supposes a considerable saving of time. It also becomes of great value especially for researchers with limited knowledge and experience in this field, but whose research topic requires the application of histological procedures to keep moving forward in their investigation.
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Different staining procedures have been performed depending on the target structures or compounds to be observed. There is a huge variety of available stains in botanical microtechnique, but hematoxylin still remains the standard nuclear stain for histological studies. However, hematoxylin by itself is a very weak dye and is of no value in microtechnique if it is not used in conjunction with a mordant that causes it to act as a very strong basic dye. Safranin O and fast green FCF staining combination, easier to apply than hematoxylin, is also considered one of the most valuable stains both for nuclear as well as anatomical and embryological studies and it has even replaced hematoxylin stain for routine work in some cases [40]. This staining combination has been applied in embryological studies of several plant species [9, 41, 42] and, in grapevine, as far as we know, for carpel morphogenesis, flower formation, fruit set and early stages of fruit development studies [37, 38, 43].
Microtechnique is an aggregate of methods used to prepare micro-objects for studying.[1] It is currently being employed in many fields in life science. Two well-known branches of microtechnique are botanical (plant) microtechnique and zoological (animal) microtechnique.
With respect to both plant microtechnique and animal microtechnique, four types of methods are commonly used, which are whole mounts, smears, squashes, and sections, in recent micro experiments.[2] Plant microtechnique contains direct macroscopic examinations, freehand sections, clearing, maceration, embedding, and staining.[3] Moreover, three preparation ways used in zoological micro observations are paraffin method, celloidin method, and freezing method.[4]
Some general microtechnique can be used in both plant and animal micro observation. Whole mounts, smears, squashes, and sections are four commonly used methods when preparing plant and animal specimens for specific purposes.[2]
Sections are known as thin slices need to be tested in all studies of cellular structures.[13] This technique can be used for the preparation of tissue of animals and plants.[14] For using under optical microscopy, the thickness of the material should be between above 2 and 25 micrometers. When observing under electron microscopy, sections should be from 20 to 30 nanometers.[2] Microtome can be used in sectioning of sufficiently thin slices. If the objects cannot satisfy the requirement of thickness, materials are required to be dehydrated using alcohol before section.[12] Three commonly used sectioning method are freehand section technique, paraffin method, and celloidin method.
Botanical microtechnique is an aggregate of methods providing micro visualization of gene and gene product in an entire plant.[15] Plant microtechnique is also a study providing valuable experimental information.[3] Plant microtechnique involves classical methods developed over a hundred years ago and new methods developed to expand our research scope and depth in botanical micro studies.[15] Both traditional and new micro technique is useful for experimental research, and some will have a significant influence on further study.[3] Different methods are used to prepare plant specimens, including direct macroscopic examinations, freehand sections,[16] clearing, maceration, embedding, and staining.
The zoological microtechnique is the art of the preparation for microscopic animal observation. Although many microtechniques can be used in both plant and animal micro experiments. Some methods may differ from itself when employed in different field. Three commonly used preparation ways used in zoological micro observations can be concluded as paraffin method, celloidin method, freezing method, and miscellaneous techniques.[4] 2ff7e9595c
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