Simple staining

Experiment no. 2.1

Simple staining

In a simple staining technique, a basic, cationic dye is flooded across a

sample, adding color to the cells.

Cationic dye (cationic- positively charged ions) is used because the cell wall
and cytoplasm of bacterial cells have a negative charge which makes the
positively charged dye attracted to the negatively charged cells, enhancing
the ability of the stain to stick to and color the cells.

cont...

In using this technique, it is important to note that before a sample can be

stained with a simple stain, it must be heat fixed to the slide. This kills the
bacteria, attaches the cells to the slide, and enhances the stain uptake.

Methylene blue is a classic example of a simple stain. This blue stain will
color all cells blue, making them stand out against the bright background of
the light microscope.

P. aeruginosa

enterobacter

S. Epidermidis

Learning
evaluation

1. Of what value is a simple stain?

Ans:
A simple stain, like Iodine, can make cell parts show up
that would otherwise nearly invisible since they are colorless.

2. Can dyes other than methylene blue be used for direct staining?
Ans:
Yes,dyes such as gram safranin, and gram crystal violet
can also be used for direct staining because they are also
cationic like methylene blue. Which means they are positively
charged and will react to the bacterial cell wall which is
negatively charged.

3. What would happen if no heat fixing were done? Or too much heat applied?
Ans:
Without heat fixing, you would lose your sample at the
time you will apply and wash your dye or the cells will be
wasted right off your slide. Too much heat applied will
damage the cells and you will not be able to see them under
the microscope

End