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Thursday, August 11, 2011
Method of Making Whole Mount Preparations
Method of Making Whole Mount Preparations Whole mount is a method of making preparations that will be observed with a microscope with no prior presence of the cutting process. So in this method, the preparation is observed that intact preparations either in the form of cells, tissues, organs and individuals. Images produced by whole-mount preparations is seen in intact form as when the organism is still alive so that observations can be made only limited to the morphology in general terms. Plants were observed to be small so it can fit on the glass object. While on a rather large plant can be done trimming (pruning) in order to become more neat and small. Examples of plants used to make preparations using the whole-mount preparations are moss, sorry spikes, leaves with trikoma and leaves with stomata. Poses of making whole-mount preparations in the moss plant that is first conducted the selection of individual moss which will be observed as much as ± 5 individuals. This is done as a repetition of each treatment so that if at one loop there are procedural errors, can be avoided by looping the other. Selection is based on the morphology of moss that is still intact vegetative and generative structures that can be observed that there are differences in the two structures. After the selection of individuals, then the moss is placed in FAA fixative solution. This treatment aims to kill the moss and fix the structure contained on the moss so that the structure does not change significantly after the moss is turned off. After fixed for ± 1 hour, then moss is washed 3 times using aquades. Then inserted into the moss Xylol solution. It aims to eliminate the remnants of fixative solution remaining on the outside of the body moss. Then the mold is transferred into a solution of hematoxylin dye, tryphan blue-lactophenol solution to provide color contrast is more apparent on the body parts of moss which will be observed. Then the moss was incubated for 2 days at room temperature to give sufficient time and temperature so the dye solution can really get into the body (tissue) mold. Moss then dehydrated in a series of alcohol 15%, 30%, 50%, 70%, 85%, 90%, 95%, absolute alcohol to remove the remnants of the liquid contained in the body of moss that is feared could be a bacterial growth substrate so that the moss becomes rot before observed. After the moss was transferred onto a glass object that has been smeared with a solution that serves as an adhesive Hoyer. In order to facilitate observation, Hoyer himself useful adhesive to attach the moss on the object and cover glass so that the observed object can not move cultivated at the time of the adhesive Hoyer there is no air bubbles that would interfere in the process of observation. After that Whole mount preparations are finished, ready to be observed under the microscope.
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