Gram Stain
(M.I. 1.2)
Loading…
Dr. Sopia Mukherjee
Assistant professor
17/09/2024
• Define Grams staining
• List the reagents used to perform Gram stain procedure and describe their purpose.
• Independently perform Gram's staining on the given smear using appropriate reagents and
timing
• Focus the appropriate field and observe the stained smear and differentiate between Gram-
positive and Gram-negative bacteria
• Draw a correct diagram with proper labeling and write a correct report
• List 2 examples each of Gram-positive cocci, Gram positive bacilli, Gram negative cocci
and Gram-negative bacilli
• Describe the factors affecting the Grams staining
• List the clinical applications of Gram’s staining
Stain
Impregnation
Simple stain Negative stain Differential stain
stain
Loading…
Gram stain
Acid fast stain
Albert stain
Trophozoite of Trichomonas
Q. Bacterial capsules can be demonstrated by:
a) Albert’s stain
b) India ink
c) Ziehl-Neelsen stain
d) LPCB
Heat fixation to help the cells
adhere to the slide surface
Crystal violet / methyl violet/ Gentian violet
Gram’s
iodine
Acetone /ethyl alcohol/
acetone alcohol
Loading…
Safranine/ dilute carbol
fuchsin
Q. All the following are components of the cell wall of gram-negative
bacteria EXCEPT:
a) Peptidoglycan
b) Lipopolysaccharide
c) Outer membrane
d) Teichoic acid
1. Staphylococci species1. Clostridium species. 1. Neisseria species. 1. Escherichia coli
2. Streptococci species. 2. Corynebacterium 2. Morexella species 2. Klebsiella
species. pneumoniae
3. Bacillus anthracis
Q. Which of the following is not a gram-positive coccus?
a) Pneumococcus
b) Meningococcus
c) Enterococcus
d) Staphylococcus
• Rapid presumptive diagnosis of diseases such as Bacterial meningitis
• Selection of Empirical antibiotics based on Gram stain finding
• Selection of suitable culture media based on Gram stain finding
• Screening of the quality of the clinical specimens such as sputum
• Fastidious organisms
• Anaerobic organisms
• Yeasts
● Smear not fixed or excessive heat fixation
● Amount and time of the decolorizer
● Age of the culture
● Thickness of smear
● Cell wall deficient organisms like Mycoplasma , Ureaplasma
● Spirochaetes- have insufficient dimensions to be resolved by light microscopy
● Organisms that exist within host cells like Chlamydiae cannot be stained
● Viability of the organisms cannot be determined on Gram staining
● Less number of bacteria.
[Link]
