3.

2: Staining Microscopic Specimens and Descriptions

Learning Objectives
Differentiate between simple and differential stains
Describe the unique features of commonly used stains
Explain the procedures and name clinical applications for Gram, endospore, acid-fast, negative capsule, and flagella
staining
What to describe about cells under the microscope

Clinical Focus: part 2

Wound infections like Cindy’s can be caused by many different types of bacteria, some of which can spread rapidly with
serious complications. Identifying the specific cause is very important to select a medication that can kill or stop the growth of
the bacteria.
After calling a local doctor about Cindy’s case, the camp nurse sends the sample from the wound to the closest medical
laboratory. Unfortunately, since the camp is in a remote area, the nearest lab is small and poorly equipped. A more modern lab
would likely use other methods to culture, grow, and identify the bacteria, but in this case, the technician decides to make a wet
mount from the specimen and view it under a brightfield microscope. In a wet mount, a small drop of water is added to the
slide, and a cover slip is placed over the specimen to keep it in place before it is positioned under the objective lens.
Under the brightfield microscope, the technician can barely see the bacteria cells because they are nearly transparent against
the bright background. To increase contrast, the technician inserts an opaque light stop above the illuminator. The resulting
darkfield image clearly shows that the bacteria cells are spherical and grouped in clusters, like grapes.

Exercise 3.2.1
1. Why is it important to identify the shape and growth patterns of cells in a specimen?
2. What other types of microscopy could be used effectively to view this specimen?

In their natural state, most of the cells and microorganisms that we observe under the microscope lack color and contrast. This
makes it difficult, if not impossible, to detect important cellular structures and their distinguishing characteristics without
artificially treating specimens. We have already alluded to certain techniques involving stains and fluorescent dyes, and in this
section we will discuss specific techniques for sample preparation in greater detail. Indeed, numerous methods have been
developed to identify specific microbes, cellular structures, DNA sequences, or indicators of infection in tissue samples, under the
microscope. Here, we will focus on the most clinically relevant techniques.

Preparing Specimens for Light Microscopy

In clinical settings, light microscopes are the most commonly used microscopes. There are two basic types of preparation used to
view specimens with a light microscope: wet mounts and fixed specimens.
The simplest type of preparation is the wet mount, in which the specimen is placed on the slide in a drop of liquid. Some
specimens, such as a drop of urine, are already in a liquid form and can be deposited on the slide using a dropper. Solid specimens,
such as a skin scraping, can be placed on the slide before adding a drop of liquid to prepare the wet mount. Sometimes the liquid
used is simply water, but often stains are added to enhance contrast. Once the liquid has been added to the slide, a coverslip is
placed on top and the specimen is ready for examination under the microscope.
The second method of preparing specimens for light microscopy is fixation. The “fixing” of a sample refers to the process of
attaching cells to a slide. Fixation is often achieved either by heating (heat fixing) or chemically treating the specimen. In addition
to attaching the specimen to the slide, fixation also kills microorganisms in the specimen, stopping their movement and metabolism
while preserving the integrity of their cellular components for observation.
To heat-fix a sample, a thin layer of the specimen is spread on the slide (called a smear), and the slide is then briefly heated over a
heat source (Figure 3.2.1). Chemical fixatives are often preferable to heat for tissue specimens. Chemical agents such as acetic

Access for free at OpenStax 3.2.1 https://bio.libretexts.org/@go/page/31773

acid, ethanol, methanol, formaldehyde (formalin), and glutaraldehyde can denature proteins, stop biochemical reactions, and
stabilize cell structures in tissue samples (Figure 3.2.1).

Figure 3.2.1 : (a) A specimen can be heat-fixed by using a slide warmer like this one. (b) Another method for heat-fixing a
specimen is to hold a slide with a smear over a microincinerator. (c) This tissue sample is being fixed in a solution of formalin (also
known as formaldehyde). Chemical fixation kills microorganisms in the specimen, stopping degradation of the tissues and
preserving their structure so that they can be examined later under the microscope. (credit a: modification of work by Nina Parker;
credit b: modification of work by Nina Parker; credit c: modification of work by “University of Bristol”/YouTube)
In addition to fixation, staining is almost always applied to color certain features of a specimen before examining it under a light
microscope. Stains, or dyes, contain salts made up of a positive ion and a negative ion. Depending on the type of dye, the positive
or the negative ion may be the chromophore (the colored ion); the other, uncolored ion is called the counterion. If the chromophore
is the positively charged ion, the stain is classified as a basic dye; if the negative ion is the chromophore, the stain is considered an
acidic dye.
Dyes are selected for staining based on the chemical properties of the dye and the specimen being observed, which determine how
the dye will interact with the specimen. In most cases, it is preferable to use a positive stain, a dye that will be absorbed by the cells
or organisms being observed, adding color to objects of interest to make them stand out against the background. However, there are
scenarios in which it is advantageous to use a negative stain, which is absorbed by the background but not by the cells or organisms
in the specimen. Negative staining produces an outline or silhouette of the organisms against a colorful background (Figure 3.2.2).

Figure 3.2.2 : (a) These Bacillus anthracis cells have absorbed crystal violet, a basic positive stain. (b) This specimen of
Spinoloricus, a microscopic marine organism, has been stained with rose bengal, a positive acidic stain. (c) These B. megaterium
appear to be white because they have not absorbed the negative red stain applied to the slide. (credit a: modification of work by
Centers for Disease Control and Prevention; credit b: modification of work by Roberto Danovaro, Antonio Pusceddu, Cristina
Gambi, Iben Heiner, Reinhardt Mobjerg Kristensen; credit c: modification of work by Anh-Hue Tu)
Because cells typically have negatively charged cell walls, the positive chromophores in basic dyes tend to stick to the cell walls,
making them positive stains. Thus, commonly used basic dyes such as basic fuchsin, crystal violet, malachite green, methylene
blue, and safranin typically serve as positive stains. On the other hand, the negatively charged chromophores in acidic dyes are
repelled by negatively charged cell walls, making them negative stains. Commonly used acidic dyes include acid fuchsin, eosin,
and rose bengal.

Access for free at OpenStax 3.2.2 https://bio.libretexts.org/@go/page/31773

Some staining techniques involve the application of only one dye to the sample; others require more than one dye. In simple
staining, a single dye is used to emphasize particular structures in the specimen. A simple stain will generally make all of the
organisms in a sample appear to be the same color, even if the sample contains more than one type of organism. In contrast,
differential staining distinguishes organisms based on their interactions with multiple stains. In other words, two organisms in a
differentially stained sample may appear to be different colors. Differential staining techniques commonly used in clinical settings
include Gram staining, acid-fast staining, endospore staining, flagella staining, and capsule staining.

Exercise 3.2.2

1. Explain why it is important to fix a specimen before viewing it under a light microscope.
2. What types of specimens should be chemically fixed as opposed to heat-fixed?
3. Why might an acidic dye react differently with a given specimen than a basic dye?
4. Explain the difference between a positive stain and a negative stain.
5. Explain the difference between simple and differential staining.

Gram Staining
The Gram stain procedure is a differential staining procedure that involves multiple steps. It was developed by Danish
microbiologist Hans Christian Gram in 1884 as an effective method to distinguish between bacteria with different types of cell
walls, and even today it remains one of the most frequently used staining techniques. The steps of the Gram stain procedure
are illustrated in Figure 3.2.3 and listed below.

Figure 3.2.3 : Gram-staining is a differential staining technique that uses a primary stain and a secondary counterstain to distinguish
between gram-positive and gram-negative bacteria.
1. First, crystal violet, a primary stain, is applied to a heat-fixed smear, giving all of the cells a purple color.
2. Next, Gram’s iodine, a mordant, is added. A mordant is a substance used to set or stabilize stains or dyes; in this case, Gram’s
iodine acts like a trapping agent that complexes with the crystal violet, making the crystal violet–iodine complex clump and
stay contained in thick layers of peptidoglycan in the cell walls.
3. Next, a decolorizing agent is added, usually ethanol or an acetone/ethanol solution. Cells that have thick peptidoglycan layers in
their cell walls are much less affected by the decolorizing agent; they generally retain the crystal violet dye and remain purple.

Access for free at OpenStax 3.2.3 https://bio.libretexts.org/@go/page/31773

 However, the decolorizing agent more easily washes the dye out of cells with thinner peptidoglycan layers, making them again
colorless.
4. Finally, a secondary counterstain, usually safranin, is added. This stains the decolorized cells pink and is less noticeable in the
cells that still contain the crystal violet dye.
The purple, crystal-violet stained cells are referred to as gram-positive cells, while the red, safranin-dyed cells are gram-negative
(Figure 3.2.4). However, there are several important considerations in interpreting the results of a Gram stain. First, older bacterial
cells may have damage to their cell walls that causes them to appear gram-negative even if the species is gram-positive. Thus, it is
best to use fresh bacterial cultures for Gram staining. Second, errors such as leaving on decolorizer too long can affect the results.
In some cases, most cells will appear gram-positive while a few appear gram-negative (as in Figure 3.2.4). This suggests damage
to the individual cells or that decolorizer was left on for too long; the cells should still be classified as gram-positive if they are all
the same species rather than a mixed culture.
Besides their differing interactions with dyes and decolorizing agents, the chemical differences between gram-positive and gram-
negative cells have other implications with clinical relevance. For example, Gram staining can help clinicians classify bacterial
pathogens in a sample into categories associated with specific properties. Gram-negative bacteria tend to be more resistant to
certain antibiotics than gram-positive bacteria. We will discuss this and other applications of Gram staining in more detail in later
chapters.

Figure 3.2.4 : In this specimen, the gram-positive bacterium Staphylococcus aureus retains crystal violet dye even after the
decolorizing agent is added. Gram-negative Escherichia coli, the most common Gram stain quality-control bacterium, is
decolorized, and is only visible after the addition of the pink counterstain safranin. (credit: modification of work by Nina Parker)

Exercise 3.2.3

1. Explain the role of Gram’s iodine in the Gram stain procedure.

2. Explain the role of alcohol in the Gram stain procedure.
3. What color are gram-positive and gram-negative cells, respectively, after the Gram stain procedure?

Clinical Focus: Part 3

Viewing Cindy’s specimen under the darkfield microscope has provided the technician with some important clues about the
identity of the microbe causing her infection. However, more information is needed to make a conclusive diagnosis. The
technician decides to make a Gram stain of the specimen. This technique is commonly used as an early step in identifying
pathogenic bacteria. After completing the Gram stain procedure, the technician views the slide under the brightfield
microscope and sees purple, grape-like clusters of spherical cells (Figure 3.2.5).

Access for free at OpenStax 3.2.4 https://bio.libretexts.org/@go/page/31773

 Figure 3.2.5: Cindy's specimen (credit: modification of work by American Society for Microbiology)

Exercise 3.2.4

1. Are these bacteria gram-positive or gram-negative?

2. What does this reveal about their cell walls?

Acid-Fast Stains
Acid-fast staining is another commonly used, differential staining technique that can be an important diagnostic tool. An acid-fast
stain is able to differentiate two types of gram-positive cells: those that have waxy mycolic acids in their cell walls, and those that
do not. Two different methods for acid-fast staining are the Ziehl-Neelsen technique and the Kinyoun technique. Both use carbol-
fuchsin as the primary stain. The waxy, acid-fast cells retain the carbol-fuchsin even after a decolorizing agent (an acid-alcohol
solution) is applied. A secondary counterstain, methylene blue, is then applied, which renders non–acid-fast cells blue.
The fundamental difference between the two carbol-fuchsin-based methods is whether heat is used during the primary staining
process. The Ziehl-Neelsen method uses heat to infuse the carbol-fuchsin into the acid-fast cells, whereas the Kinyoun method does
not use heat. Both techniques are important diagnostic tools because a number of specific diseases are caused by acid-fast bacteria
(AFB). If AFB are present in a tissue sample, their red or pink color can be seen clearly against the blue background of the
surrounding tissue cells (Figure 3.2.6).

Exercise 3.2.5

Why are acid-fast stains useful?

Using Microscopy to Diagnose Tuberculosis

Mycobacterium tuberculosis, the bacterium that causes tuberculosis, can be detected in specimens based on the presence of
acid-fast bacilli. Often, a smear is prepared from a sample of the patient’s sputum and then stained using the Ziehl-Neelsen
technique (Figure 3.2.6). If acid-fast bacteria are confirmed, they are generally cultured to make a positive identification.
Variations of this approach can be used as a first step in determining whether M. tuberculosis or other acid-fast bacteria are
present, though samples from elsewhere in the body (such as urine) may contain other Mycobacterium species.
An alternative approach for determining the presence of M. tuberculosis is immunofluorescence. In this technique,
fluorochrome-labeled antibodies bind to M. tuberculosis, if present. Antibody-specific fluorescent dyes can be used to view the
mycobacteria with a fluorescence microscope.

Access for free at OpenStax 3.2.5 https://bio.libretexts.org/@go/page/31773

 Figure 3.2.6 : Ziehl-Neelsen staining has rendered these Mycobacterium tuberculosis cells red and the surrounding growth
indicator medium blue. (credit: modification of work by American Society for Microbiology)

Capsule Staining
Certain bacteria and yeasts have a protective outer structure called a capsule. Since the presence of a capsule is directly related to a
microbe’s virulence (its ability to cause disease), the ability to determine whether cells in a sample have capsules is an important
diagnostic tool. Capsules do not absorb most basic dyes; therefore, a negative staining technique (staining around the cells) is
typically used for capsule staining. The dye stains the background but does not penetrate the capsules, which appear like halos
around the borders of the cell. The specimen does not need to be heat-fixed prior to negative staining.
One common negative staining technique for identifying encapsulated yeast and bacteria is to add a few drops of India ink or
nigrosin to a specimen. Other capsular stains can also be used to negatively stain encapsulated cells (Figure 3.2.7). Alternatively,
positive and negative staining techniques can be combined to visualize capsules: The positive stain colors the body of the cell, and
the negative stain colors the background but not the capsule, leaving halo around each cell.

Figure 3.2.7 : (a) India-ink was used to stain the background around these cells of the yeast Cryptococcus neoformans. The halos
surrounding the cells are the polysaccharide capsules. (b) Crystal violet and copper sulfate dyes cannot penetrate the encapsulated
Bacillus cells in this negatively stained sample. Encapsulated cells appear to have a light-blue halo. (credit a: modification of work
by American Society for Microbiology; credit b: modification of work by American Society for Microbiology)

Exercise 3.2.6

How does negative staining help us visualize capsules?

Endospore Staining
Endospores are structures produced within certain bacterial cells that allow them to survive harsh conditions. Gram staining alone
cannot be used to visualize endospores, which appear clear when Gram-stained cells are viewed. Endospore staining uses two
stains to differentiate endospores from the rest of the cell. The Schaeffer-Fulton method (the most commonly used endospore-
staining technique) uses heat to push the primary stain (malachite green) into the endospore. Washing with water decolorizes the
cell, but the endospore retains the green stain. The cell is then counterstained pink with safranin. The resulting image reveals the
shape and location of endospores, if they are present. The green endospores will appear either within the pink vegetative cells or as

Access for free at OpenStax 3.2.6 https://bio.libretexts.org/@go/page/31773

separate from the pink cells altogether. If no endospores are present, then only the pink vegetative cells will be visible (Figure
3.2.8).

Figure 3.2.8 : A stained preparation of Bacillus subtilis showing endospores as green and the vegetative cells as pink. (credit:
modification of work by American Society for Microbiology)
Endospore-staining techniques are important for identifying Bacillus and Clostridium, two genera of endospore-producing bacteria
that contain clinically significant species. Among others, B. anthracis (which causes anthrax) has been of particular interest
because of concern that its spores could be used as a bioterrorism agent. C. difficile is a particularly important species responsible
for the typically hospital-acquired infection known as “C. diff.”

Exercise 3.2.7

Is endospore staining an example of positive, negative, or differential staining?

Flagella Staining
Flagella (singular: flagellum) are tail-like cellular structures used for locomotion by some bacteria, archaea, and eukaryotes.
Because they are so thin, prokaryote flagella typically cannot be seen under a light microscope without a specialized flagella
staining technique. Flagella staining thickens the flagella by first applying mordant (generally tannic acid, but sometimes potassium
alum), which coats the flagella; then the specimen is stained with pararosaniline (most commonly) or basic fuchsin (Figure 3.2.9).

Figure 3.2.9 : A flagella stain of Bacillus cereus, a common cause of foodborne illness, reveals that the cells have numerous
flagella, used for locomotion. (credit: modification of work by Centers for Disease Control and Prevention)
Though flagella staining is uncommon in clinical settings, the technique is commonly used by microbiologists, since the location
and number of flagella can be useful in classifying and identifying bacteria in a sample. When using this technique, it is important
to handle the specimen with great care; flagella are delicate structures that can easily be damaged or pulled off, compromising
attempts to accurately locate and count the number of flagella.

Exercise 3.2.8

Is the flagella stain showing the true size of the flagella?

Access for free at OpenStax 3.2.7 https://bio.libretexts.org/@go/page/31773

Figure 3.2.10 : (credit “basic stains”: modification of work by Centers for Disease Control and Prevention; credit “Acidic stains”:
modification of work by Roberto Danovaro, Antonio Dell’Anno, Antonio Pusceddu, Cristina Gambi, Iben Heiner, Reinhardt
Mobjerg Kristensen; credit “Negative stains”: modification of work by Anh-Hue Tu)

Figure 3.2.11 : (credit “Gram stain”: modification of work by Nina Parker; credit “Acid-fast stain”: modification of work by
American Society for Microbiology; credit “Endospore stain”: modification of work by American Society for Microbiology; credit
“Capsule stain” : modification of work by American Society for Microbiology; credit “Flagella stain”: modification of work by
Centers for Disease Control and Prevention)

Access for free at OpenStax 3.2.8 https://bio.libretexts.org/@go/page/31773

Preparing Specimens for Electron Microscopy
Samples to be analyzed using a TEM must have very thin sections. But cells are too soft to cut thinly, even with diamond knives.
To cut cells without damage, the cells must be embedded in plastic resin and then dehydrated through a series of soaks in ethanol
solutions (50%, 60%, 70%, and so on). The ethanol replaces the water in the cells, and the resin dissolves in ethanol and enters the
cell, where it solidifies. Next, thin sections are cut using a specialized device called an ultramicrotome (Figure 3.2.12). Finally,
samples are fixed to fine copper wire or carbon-fiber grids and stained—not with colored dyes, but with substances like uranyl
acetate or osmium tetroxide, which contain electron-dense heavy metal atoms.

Figure 3.2.12 : (a) An ultramicrotome used to prepare specimens for a TEM. (b) A technician uses an ultramicrotome to slice a
specimen into thin sections. (credit a: modification of work by “Frost Museum”/Flickr; credit b: modification of work by U.S. Fish
and Wildlife Service Northeast Region)
When samples are prepared for viewing using an SEM, they must also be dehydrated using an ethanol series. However, they must
be even drier than is necessary for a TEM. Critical point drying with inert liquid carbon dioxide under pressure is used to displace
the water from the specimen. After drying, the specimens are sputter-coated with metal by knocking atoms off of a palladium
target, with energetic particles. Sputter-coating prevents specimens from becoming charged by the SEM’s electron beam.

Exercise 3.2.9

1. Why is it important to dehydrate cells before examining them under an electron microscope?
2. Name the device that is used to create thin sections of specimens for electron microscopy.

Using Microscopy to Diagnose Syphilis

Figure 3.2.13 : (a) Living, unstained Treponema pallidum spirochetes can be viewed under a darkfield microscope. (b) In this
brightfield image, a modified Steiner silver stain is used to visualized T. pallidum spirochetes. Though the stain kills the cells,
it increases the contrast to make them more visible. (c) While not used for standard diagnostic testing, T. pallidum can also be
examined using scanning electron microscopy. (credit a: modification of work by Centers for Disease Control and Prevention;
credit b: modification of work by Centers for Disease Control and Prevention; credit c: modification of work by Centers for
Disease Control and Prevention)
The causative agent of syphilis is Treponema pallidum, a flexible, spiral cell (spirochete) that can be very thin (<0.15 μm) and
match the refractive index of the medium, making it difficult to view using brightfield microscopy. Additionally, this species
has not been successfully cultured in the laboratory on an artificial medium; therefore, diagnosis depends upon successful
identification using microscopic techniques and serology (analysis of body fluids, often looking for antibodies to a pathogen).
Since fixation and staining would kill the cells, darkfield microscopy is typically used for observing live specimens and
viewing their movements. However, other approaches can also be used. For example, the cells can be thickened with silver

Access for free at OpenStax 3.2.9 https://bio.libretexts.org/@go/page/31773

 particles (in tissue sections) and observed using a light microscope. It is also possible to use fluorescence or electron
microscopy to view Treponema (Figure 3.2.13).
In clinical settings, indirect immunofluorescence is often used to identify Treponema. A primary, unstained antibody attaches
directly to the pathogen surface, and secondary antibodies “tagged” with a fluorescent stain attach to the primary antibody.
Multiple secondary antibodies can attach to each primary antibody, amplifying the amount of stain attached to each Treponema
cell, making them easier to spot (Figure 3.2.14).

Figure 3.2.14 : Indirect immunofluorescence can be used to identify T. pallidum, the causative agent of syphilis, in a specimen.

Preparation and Staining for Other Microscopes

Samples for fluorescence and confocal microscopy are prepared similarly to samples for light microscopy, except that the dyes are
fluorochromes. Stains are often diluted in liquid before applying to the slide. Some dyes attach to an antibody to stain specific
proteins on specific types of cells (immunofluorescence); others may attach to DNA molecules in a process called fluorescence in
situ hybridization (FISH), causing cells to be stained based on whether they have a specific DNA sequence. Sample preparation for
two-photon microscopy is similar to fluorescence microscopy, except for the use of infrared dyes.

Exercise 3.2.10

What is the main difference between preparing a sample for fluorescence microscopy versus light microscopy?

Cornell University’s Case Studies in Microscopy offers a series of clinical problems based on real-life events. Each case study
walks you through a clinical problem using appropriate techniques in microscopy at each step.

Microscopy and Antibiotic Resistance

As the use of antibiotics has proliferated in medicine, as well as agriculture, microbes have evolved to become more resistant.
Strains of bacteria such as methicillin-resistant S. aureus (MRSA), which has developed a high level of resistance to many
antibiotics, are an increasingly worrying problem, so much so that research is underway to develop new and more diversified
antibiotics.
Fluorescence microscopy can be useful in testing the effectiveness of new antibiotics against resistant strains like MRSA. In a
test of one new antibiotic derived from a marine bacterium, MC21-A (bromophene), researchers used the fluorescent dye
SYTOX Green to stain samples of MRSA. SYTOX Green is often used to distinguish dead cells from living cells, with
fluorescence microscopy. Live cells will not absorb the dye, but cells killed by an antibiotic will absorb the dye, since the
antibiotic has damaged the bacterial cell membrane. In this particular case, MRSA bacteria that had been exposed to MC21-A
did, indeed, appear green under the fluorescence microscope, leading researchers to conclude that it is an effective antibiotic
against MRSA.
Of course, some argue that developing new antibiotics will only lead to even more antibiotic-resistant microbes, so-called
superbugs that could spawn epidemics before new treatments can be developed. For this reason, many health professionals are
beginning to exercise more discretion in prescribing antibiotics. Whereas antibiotics were once routinely prescribed for
common illnesses without a definite diagnosis, doctors and hospitals are much more likely to conduct additional testing to
determine whether an antibiotic is necessary and appropriate before prescribing.
A sick patient might reasonably object to this stingy approach to prescribing antibiotics. To the patient who simply wants to
feel better as quickly as possible, the potential benefits of taking an antibiotic may seem to outweigh any immediate health
risks that might occur if the antibiotic is ineffective. But at what point do the risks of widespread antibiotic use supersede the
desire to use them in individual cases?

Access for free at OpenStax 3.2.10 https://bio.libretexts.org/@go/page/31773

Description of Cells Under the Microscope
Once you have stained the cells, the next step is to use the microscope to describe them. You will always look for cell shape, the
grouping and the color(s) that appear. These descriptions of cell morphology (shape and structure) are your microscopic
observations. This is most commonly done with prokaryotes and the following tables contain proper names for shapes and
groupings you should use.

Figure 3.2.15: (credit “Coccus” micrograph: modification of work by Janice Haney Carr, Centers for Disease Control and
Prevention; credit “Coccobacillus” micrograph: modification of work by Janice Carr, Centers for Disease Control and Prevention;
credit “Spirochete” micrograph: modification of work by Centers for Disease Control and Prevention)

Access for free at OpenStax 3.2.11 https://bio.libretexts.org/@go/page/31773

 Figure 3.2.16 : Common cell arrangements.
Eukaryotic cells display a wide variety of different cell morphologies. Possible shapes include spheroid, ovoid, cuboidal,
cylindrical, flat, lenticular, fusiform, discoidal, crescent, ring stellate, and polygonal (Figure 3.2.17). Some eukaryotic cells are
irregular in shape, and some are capable of changing shape. The shape of a particular type of eukaryotic cell may be influenced by
factors such as its primary function, the organization of its cytoskeleton, the viscosity of its cytoplasm, the rigidity of its cell
membrane or cell wall (if it has one), and the physical pressure exerted on it by the surrounding environment and/or adjoining cells.

Figure 3.2.17 : Eukaryotic cells come in a variety of cell shapes. (a) Spheroid Chromulina alga. (b) Fusiform shaped Trypanosoma.
(c) Bell-shaped Vorticella. (d) Ovoid Paramecium. (e) Ring-shaped Plasmodium ovale. (credit a: modification of work by NOAA;
credit b, e: modification of work by Centers for Disease Control and Prevention).

Access for free at OpenStax 3.2.12 https://bio.libretexts.org/@go/page/31773

 Exercise 3.2.11

1. What are the proper names for the common shapes of bacterial cells?
2. What are the proper names for the common groupings of bacterial cells?
3. Why do eukaryote cells not always follow the same shapes and groupings as bacterial cells?

Key Concepts and Summary

Samples must be properly prepared for microscopy. This may involve staining, fixation, and/or cutting thin sections.
A variety of staining techniques can be used with light microscopy, including Gram staining, acid-fast staining, capsule
staining, endospore staining, and flagella staining.
Samples for TEM require very thin sections, whereas samples for SEM require sputter-coating.
Preparation for fluorescence microscopy is similar to that for light microscopy, except that fluorochromes are used.
Cells have several things unique about them when being viewed under the microscope. When recording observations, make
sure to describe cell shape, grouping and color(s).

Contributor
Nina Parker, (Shenandoah University), Mark Schneegurt (Wichita State University), Anh-Hue Thi Tu (Georgia Southwestern
State University), Philip Lister (Central New Mexico Community College), and Brian M. Forster (Saint Joseph’s University)
with many contributing authors. Original content via Openstax (CC BY 4.0; Access for free
at https://openstax.org/books/microbiology/pages/1-introduction)

This page titled 3.2: Staining Microscopic Specimens and Descriptions is shared under a CC BY license and was authored, remixed, and/or
curated by OpenStax.

Access for free at OpenStax 3.2.13 https://bio.libretexts.org/@go/page/31773