Microscopic and counting techniques
The counting and identification of spores and pollen grains should be done through light microscope (LM). It is usually possible and convenient to identify and count samples from prepared slides by scanning at a magnification of x400 (12, 19). This provides sufficient magnification for routine identification, and has advantage that the field of view resulting from the use of x40 objective lens is usually appropriate for slides with a reasonable pollen density. Additionally, the use of x100 objective lens has to be recommended for ambiguous grains.
The number of pollen and spore should be counted in order to represent the pollen sum. Commonly counting pollen for analysis requires at least 200 pollen grains per sample (13). Nevertheless, it should be counted as much as possible. In this study, pollens and spores should be counted at least 200 grains from 4 slides per sample to calculate as percentage to obtain the proportion of pollen taxa and make a pollen diagram. However, if it can not be reached, counting result will show the result of total counting from 6 slides