Protocol for Slide Preparation for Daily Pollen Count

1) Remove tape (exposed for previous 24 hours) from drum and align on perspex ruled block.

2) Cut 24 hour exposed section and transfer carefully to clean, dated slide. Placing a small drop of distilled water in the middle of the slide prior to lowering the tape will help the latter to adhere during inversion of slide.

Wind vane Lid assembly

Rain shield

Orifice Rotation lock

Motor cover

Wind vane Lid assembly

Rain shield

Orifice Rotation lock

Motor cover

Fig. 13.8-a Burkard seven-day recording volumetric spore trap.

Trapping Surface

Trapping Surface

Lid Start reference pointer lock nut

Orifice start position

Fig. 13.8b Burkard seven-day recording volumetric spore trap showing details of lid with clock-driven drum with adhesive-coated tape.

3) Meantime phenolyzed basic fuchsin stain has to be kept for melting on a hot plate.

4) Transfer sufficient melted stain to cover area of cover slip when placed on slide. Pasteur pipette is used for this (cover slips are 20 x 60 mm No. 1 thickness).

5) Invert slide with tape onto cover slip bearing stain by means of a mounted needle. Apply gentle pressure to cover slip having inverted the slide with cover slip + tape on upper surface.

6) Place slide on microscope stage with date and start of exposure period on the left of the stage, at mid-point of the slide.

7) Move the slide to where the edge of the tape is aligned with the right hand of the microscope field.

8) Note the position on the horizontal vernier scale.

9) Move the tape 2 mm on to begin the first transect.

10) Move the slide vertically across the field examining x 400 magnification (x 10 eyepiece in conjunction with x 40 objective).

11) Identify and count each pollen grain as it appears. Grains are counted even if only partially visible in the left hand margin of the field, but not counted if partially covered in the right hand margin of the field (i.e., polar halves).

12) Record each taxon [usually at generic level, but for grasses at family (Poaceae) level].

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