2. Remove a small amount of the culture from the slant with an inoculating loop and disperse it in the drop of stain without spreading the drop.
3. Use another clean slide to spread the drop of stain containing the
organism using the following technique.
4. Rest one end of the clean slide on the center of the slide with the
stain. Tilt the clean slide toward the drop forming an acute angle
and draw that slide toward the drop until it touches the drop
and causes it to spread along the edge of the spreader slide. Maintaining
a small acute angle between the slides, push the spreader slide toward
the clean end of the slide being stained dragging the drop behind the spreader
slide and producing a broad, even, thin smear.
5. Allow the smear to dry without heating.
6. Focus a thin area under oil immersion and observe the unstained cells surrounded by the gray stain.
Results of a Negative stain