Why perform staining process on micro-organism ?
Staining on micro-organisms using dye for more clearly identify morphological characteristics , components of microorganisms by Enhanced the contrast in samples.
Identification of Bacteria
After isolation of pure culture. it needs to be indentified Morphology, characteristics of media, components of cell membrane, resistance, metabolism via different techniques.
SIMPLE STAINING
In which bacterial smear stained by using a Single stain dye {Methylene blue, safranine, crystal violet dye}.
This staining process is used for indentification of Morphological characteristics { shape and size} of microorganisms (bacteria).
That which does not differentiate micro-organisms.
Principle:-
Bacterial colonies (Negatively charged) stained by Positive charged dye .
So dye attached easily on the bacterial surface due to -+ charge attraction.
So bacteria visible ( high intensity with light background) under microscopy.
Generated negative charge on bacterial colonies by adding of OH- (Hydroxide ion) or remove of H+ (hydrogen ion).
Procedure/process:-
👉 Firstly slide is taken
👉 Slide cleaned/washed with detergents then air dry and sterilize it.
👉 After sterilization makes smear of bacterial suspension on slide by using sterile inoculating loop .
👉 Then smear slide allow to air dry , for heat fixation on smear slide, it passes over the flame for 3-4 times .
👉 After heat fixation smear staining by few drops of methylene blue dye.
👉 Then allow to air dry (few times).
👉 After drying smear slide washed excess dye under distilled water.
👉 Then, now slowly wiping the smear slide with tissue paper.
👉 Then smear slide covered with coverslip.
👉 Now bacterial slide(smear) ready to observe the bacteria under microscopy.
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Thank you.
👉👉Part-2 Differential Staining
👉Also read other microbiology topics
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