Bacillus Spe Lab Report

The prepared slides of Bacillus sp. were placed on the microscope stage and by using 4X and then 10X objective, focus the microscope on an appropriate field containing bacteria. Centre the bacteria in the field of view, by manipulating the lateral movement knob. Continue rotate the nose piece to the 40X objectives and use fine focus to refocus. The next objectives lens is 100X. Place a small drop of immersion oil when the nose piece at the position between the 40X and 100X objective is obtained, so that the 100X objectives is rotated into the oil. The shape of bacteria were draw. Clean the immersion oil from 100X by rubbing it with the lens paper dipped into the alcohol, xylol or toluene.

Ocular micrometre is the simple matter to measure the

Examine the smear under microscope from 4x objective until 100x objective. Draw the cell, shape, arrangement and colour of cells.

For negative staining, culture of Klebsiella were used. A small drop of nigrosine were place on the slide and after that place the culture from the slant in the drop of stain. Spread the drop of stain with another clean slide by push the spreader slide towards the clean end of the slide being stained dragging the drop behind the spreader slide and producing a broad, even and thin smear. Allow it to dry and examine it under the microscope.

The heat-fixed smear of E. coli sp. and Staphylococcus sp. were prepared for the Negative staining. Placed the fixed smear on the staining rack and flooding with the crystal violet for 1 minute. By using the tab water, rinse the slide. After that, by using Lugol’s Iodine, cover the whole slide for 1 minute. The washing step were repeated. Using (95% v/v) alcohol wash off the iodine solution until the washing are pale violet. Immediately rinse with tab water. Using the Safranin solution (0.5% w/v), cover the smear for 1 minute. Use tab water to rinse off the stain. By using paper towel, blot dry the smear and observed under the

were used for the Endospore staining. The smear prepared by air-dry and heat-fixed. Boiled the water in the beaker the hot plate. Place the wire stain rack on the beaker which has steam coming up from boiled water. A small piece of paper towel were placed on top of smear to keep dye from evaporating to fast and giving more contact time between the dye and the bacterial cell walls. Using Malachite green and flood the smear and leave it for 5 minutes in the fumes hood cupboard. The piece of paper towel were removed. Wash really well with the water. Flood the smear using Safranin for 1 minutes. Wash the stain. Observed the smear under the microscope.

Klebsiella sp. were used for the Capsular staining. Small amount of bacterium were mix aseptically with wire loop and after that flame the loop. Placed the slide on the staining rack and mount the crystal violet. Heat the slide slowly under the flame a few times and let it to air-dry for 1 minute. Wash the crystal violet with tab water. Using the paper towel to dry the slide slowly and allow it to air- dry. Examine the smear under the