EXERCISE NO.

4
Staining Techniques

Answer Sheets

Name: RHYN RUZ G. DOHILAG Rating: __________

Laboratory Section: BIO117 W67A78 Date: SEPT. 17, 2021

A. Observations. Draw the cells you have observed based on the demonstrations.

Illustration Description

Methylene
blue stains negatively
charged molecules in
the cell, including DNA
and RNA.
Cheek cells are
eukaryotic cells (cells
that contain a nucleus
and other organelles
within enclosed in a
membrane) that are
easily shed from the
mouth lining. It's
therefore easy to obtain
them for observation.

Figure 1. Simple Stain

The Gram stain

procedure distinguishes
between Gram positive
and Gram negative
groups by coloring
these cells red or violet.

It is a common technique
used to differentiate two
large groups of bacteria
based on their different
cell wall constituents.

Figure 2. Gram Stain

1. Using the CF you obtained for 40X objective lens from Ex. 3, determine the size of the
cheek cell below:

CF (40X) = low power magnification

CF (10X) high power magnification

CF (40X) = 100X
8 400X

CF (40X) = 100X (8)

400X

CF (40X) = 2

Cheek Cell Size = 25 divisions x 2 = 50 µm

Guide questions:

1. What are the differences between simple staining and differential staining?

- The simple staining will only tell you morphological shape and size, while the
differential staining will allow for determination of size, shape, and type of the cell
wall.

- Simple staining involves directly staining the bacterial cell with a positively

charged dye. While, differential staining distinguishes organisms based on their
interactions with multiple stains.

2. Identify the importance of each step in Gram staining.

- The Gram stain is a very important preliminary step in the initial characterization
and classification of bacteria. It is also a key procedure in the identification of
bacteria based on staining characteristics, enabling the bacteria to be examined
using a light microscope.
- The performance of the Gram Stain on any sample requires four basic steps that
include applying a primary stain (crystal violet) to a heat-fixed smear, followed by
the addition of a mordant (Gram’s Iodine), rapid decolorization with alcohol,
acetone, or a mixture of alcohol and acetone and lastly, counterstaining with
safranin.