Bacteria Classification By Gram Staining Essay Research

Bacteria Classification By Gram Staining Essay, Research Paper

Bacteria Classification By Gram Staining

THE AMERICAN UNIVERSITY IN CAIRO

BIOLOGY DEPARTMENT

SCIENCE 453: Biology FOR ENGINEERS REPORT No.1

Presented By: Karim A. Zaklama 92-1509 Sci. 453-01

24/2/96

Aim:

To prove a sample of research lab prepared bacteriums and categorize it

harmonizing to Christian? s gm positive and gram negative categories and besides by

sing it under a high powered microscope and oil submergences ; sort its

form and note any particular features.

Introduction:

Bacteria was categorised into two groups in 1884 by the Danish

Bacteriologist Christian, gm positive and gram negative by a staining

technique where the ability to avoid de-coloration of Crystal Violet solution

by intoxicant would render the class of gm positive, and gram negative if the

bacteriums is de-coloured. This could be noted by the concluding coloring material of the

bacteriums: a violet coloring material where Gram positive and a pink coloring material of the Safranin

added pending the de-colouring procedure.

Materials:

1. Bacteria Sample 2. Microscope Slide 3. Gram Staining Kit and Wash Bottles

a. Crystal Violet Solution b. Iodine Solution c. 95 % Ethyl Alcohol vitamin D.

Safranin e. Distilled Water 4. Bibulous Blotting Paper 5. Microscope 6. Oil

Procedure:

A. Preparation:

1. Bacteria is cultivated on agar jelly in an brooder at 25? C for 24 hours. 2.

Obtain a microscope slide and with a toothpick, smear a thin coat of the

bacteriums sample onto the slide 3. Cover the smear with a bead Crystal Violet

and go forth standing for 20 seconds 4. Wash off the discoloration with distilled H2O ;

drain and smudge off the extra with boozy paper. 5. Use Gram? s Iodine on

the vilification and leave to stand for 1 minute. 6. Drain the extra I and use

95 % Ethyl intoxicant for 20 2nd continuance or till the intoxicant runs clearly from

the slide. 7. The vilification should rinsed for a few seconds with distilled H2O to

halt the action of the intoxicant. 8. Drain and smudge off the extra with boozy

9. Introduce Safranin to the vilification and leave standing for 20 seconds. 10. Wash

off the discoloration with distilled H2O ; drain and smudge off the extra with boozy

paper. 11. Leave the slide to air prohibitionists.

B. Examination:

1. Put the slide under microscope on low powered lens. 2. Travel the slide

utilizing the setup until the sample can be seen as a fuzz under the microscope.

3. F

ocus the lens to guarantee that there is a sample straight under the lens. 4.

Move to higher powered lens, repeat measure 3. 5. Travel to higher powered lens,

repeat measure 3 6. Move microscope aside and add Oil submergence, leave for a few

seconds and re-examine the slide.

Note Shape and coloring material and any other observations.

Consequences and Observations:

It was apparent by ocular scrutiny that the intoxicant was de-colouring or a

least partly de-colouring the bacterium.

The sample appeared a dark pink or shut to violet by the bare oculus ; a

microscope was needed to guarantee consequences.

Under the low powered microscope sunglassess of pink were noted.

Under the medium power, the sunglassess were more clear but no form could be made

out.

Under the high powered microscope bunchs of pink rod ( B ) shaped bacteriums

cells could be observed.

Under Oil Immersion and high powered lens the cells could seen more separated out

and therefore a clearer indicant of the pink coloring material, bacilli form and spores could

be made out in the single cells.

Decision:

The Shape was noted as Bacilli ( Rod-like ) shaped cells ; a gm variable

form, distinct in either Gram Negative or Gram positive bacteriums.

The concluding coloring material of the cells were stained pink by the Safranin screening

the de-coloration of the crystal violet turn outing the bacterium is of the gm

negative category.

Under oil submergence the cells became more thin and under the high

powered lens of the microscope spores could be seen, as small bubbles, in the

cells. This tells us that the bacterium was in its terminal province.

The presence of spores in the bacterium at its terminal province Tells us

that the bacteriums could be an old civilization. Old bacterium civilizations which are gm

positive tend to de-colour, yet more easy than gram negative bacteriums. The

velocity of de-coloration was non inspected really clearly therefore no farther decision

could be reached, yet it is possible that this an old civilization of Bacilli shaped

Gram Positive bacterium.

Recommendation:

It is recommended that the same sample be tested once more for de-

colour ; concentrating on de-coloration velocity. If the de-coloration is fast so

the sample is decidedly gram negative, slow de-coloration would state us it is

gm positive. For future samples it would be recommended to maintain the bacterium

sample for this specific trial for merely 16 hours every bit recommended to avoid the

presence of old civilizations which are anomalous to this trial.