Far Eastern University

Nicanor Reyes Street

Sampaloc, Manila

LABORATORY EXERCISE NO. 1B

BACTERIOLOGY: Bacterial Morphology and Gram Staining

Submitted by:

Camba, Alexie Justine Louise B.

Kua, Cyrine Khryssta

Magtibay, Erick Louie T.

Santing, Rica A.

Tiongco, Althea

Submitted to:

Ms. Leny Saldo

©
Biofacultymember2020

Bio1227 Microbiology and Parasitology

Department of Biology

Institute of Arts and Sciences

Far Eastern University

 Far Eastern University
Nicanor Reyes Street
Sampaloc, Manila

INTRODUCTION

We all recall our mothers warning us about germs and how they could infect us. She's

correct, of course. Have you ever thought about how microorganisms are identified? The study of

how living organisms develop is known as Morphology. (Carrillo-Lopez & Yahia, 2019) A gram

stain is used to divide bacteria into two groups depending on their cell wall construction.

Identification of bacteria is critical for disease diagnosis. Peptidoglycans are substances found in

the cell walls of both gram positive and gram negative bacteria.

METHODOLOGY

To reach the goal of this study, we used the Gram staining process to help us determine if

the bacteria were gram-positive or gram-negative. The Gram staining involves three processes:

staining with a water-soluble dye called crystal violet, decolorization, and counterstaining, usually

with safanin.

©
Biofacultymember2020

Bio1227 Microbiology and Parasitology

Department of Biology

Institute of Arts and Sciences

Far Eastern University

 Far Eastern University
Nicanor Reyes Street
Sampaloc, Manila

RESULTS AND DISCUSSIONS

Click on the following link: Search Help in Finding Bacteria Cell Labeling - Online Quiz
Version. Each member must play the game. Make as many attempts until you will perfect the
game. Print screen you final score.

Review the cytoplasmic membrane of bacteria. You may refer to the following URL:
https://www.cellsalive.com/cells/bactcell_js.htm OR search this topic on other preferred websites

TIONGCO: CAMBA:

©
Biofacultymember2020

Bio1227 Microbiology and Parasitology

Department of Biology

Institute of Arts and Sciences

Far Eastern University

 Far Eastern University
Nicanor Reyes Street
Sampaloc, Manila

MAGTIBAY: SANTING:

KUA:

©
Biofacultymember2020

Bio1227 Microbiology and Parasitology

Department of Biology

Institute of Arts and Sciences

Far Eastern University

 Far Eastern University
Nicanor Reyes Street
Sampaloc, Manila

Describe and illustrate the cytoplasmic membrane of Gram (-) and Gram (+)

Photo from Sarthaks eConnect:

https://www.sarthaks.com/885472/describe-the-structure-gram-positive-and-gram-negative-
bacterial-cell-wall-usi

Gram-negative bacteria have two membranes that sandwich a layer of peptidoglycan (diderm

bacteria), whereas Gram-positive bacteria have a thicker layer of peptidoglycan surrounding a

single membrane (monoderm bacteria).

©
Biofacultymember2020

Bio1227 Microbiology and Parasitology

Department of Biology

Institute of Arts and Sciences

Far Eastern University

 Far Eastern University
Nicanor Reyes Street
Sampaloc, Manila

Answer the following questions: (Just choose at least 5 questions to answer)

1. What is the advantage of the Gram stain over the simple stain?

● Gram stains highlight different bacteria types through use of special dyes. It aids in the

diagnosis of specific organisms. it differentiates between gram positive and gram negative

bacteria.

2. What is the theory about the mechanism of the Gram-stain reaction?

● The technique for gram staining tests the ability of the bacteria to retail crystal violet dye

after the solvent treatment or the procedure is based on the ability of the microorganism to

retain the color of stains used during gram negative and gram positive reaction.

● gram positive bacterial has thick peptidoglycan layer than the gram-negative bacteria and

thus it retains the crystal violet dye whereas gram negative bacteria consists of more lipids

in their cell wall and thus treatment of ethanol washes off the crystal violet dye and turns it

into pinkish red color after it is treated with safranin.

3. What part of the bacterial cell is most involved with Gram staining, and why?

● The most crucial step of the gram staining method is the decolorizing step where ethanol

is added to both gram negative and gram positive bacteria so that it washes off the crystal

violet dye from the gram-negative bacteria.

©
Biofacultymember2020

Bio1227 Microbiology and Parasitology

Department of Biology

Institute of Arts and Sciences

Far Eastern University

 Far Eastern University
Nicanor Reyes Street
Sampaloc, Manila

● But if the decolourizer is left for too long, it may wash off the crystal violet dye from both

gram positive and gram negative bacteria

4. What is the purpose of the alcohol solution in the Gram stain?

● It is used as a decolourizing agent that allows iodine crystal violet complexes to leak out

of thin peptidoglycan layer.

5. What counterstain is used in Gram staining? Why is it necessary? Could colors other

than Safranin red be used?

● Gram staining is not complete without counterstain because gram negative bacteria take

color of the counterstain. It is necessary because safranin is used and the basic function or

malachite green can also be used.

CONCLUSION

The Staining procedure is a convenient and quick technique which is also extremely

reliable as well as easily accessible in research facilities. As a result, the initial characterization

and segmentation of each representative of the domain, it acts as a fundamental procedure in the

bacterial identification based on subjective staining properties, where it examines specific

domains.

©
Biofacultymember2020

Bio1227 Microbiology and Parasitology

Department of Biology

Institute of Arts and Sciences

Far Eastern University

 Far Eastern University
Nicanor Reyes Street
Sampaloc, Manila

REFERENCE

Carrillo-Lopez, A., & Yahia, E.M., (2019). Morphology and Anatomy. ScienceDirect.
https://www.sciencedirect.com/topics/agricultural-and-biological-sciences/
Atlas of Oral Microbiology, 2015. Sciencedirect.com.
https://www.sciencedirect.com/topics/medicine-and-dentistry/microbial-morphology

Monica Z. Bruckner (2021) Gram Staining. serc.carleton.edu.

https://serc.carleton.edu/microbelife/research_methods/microscopy/gramstain.html

Tripathi N, Sapra A. Gram Staining. [Updated 2021 Feb 23]. In: StatPearls [Internet]. Treasure
Island (FL): StatPearls Publishing; 2021 Jan-. Available from:
https://www.ncbi.nlm.nih.gov/books/NBK562156/

©
Biofacultymember2020

Bio1227 Microbiology and Parasitology

Department of Biology

Institute of Arts and Sciences

Far Eastern University