Microscope Slides Preparation
Styles and Techniques
Using prepared microscope slides
Objects magnified under compound microscopes are mounted onto microscope slides.
Made of glass or plastic, slides are approximately 1x3 inches and between 1mm-1.2
mm thick.
Multiple methods of preparation allow for advanced viewing of inorganic and organic objects.
Flat and Concave Styles
The most basic of all microscope slides is a flat rectangular piece of soda lime glass,
borosilicate cover glass or plastic, with ground edges.
All corners are a sharp 90-degrees and, along with a rough outer edge, can cause minor
finger cuts if not handled with care.
The top and/or bottom edges of a slide can be frosted, enabling easy marking for
sample identification and/or orientation. The etched frosting keeps all pen marks safely
away from the sample and a selection of frosted colors provides additional means of
categorization.
Rounded safety corners to prevent accidental cuts as well as beveled edges with clipped corners ideal for blood
samples are options available for both generic and frosted slides.
Concave microscope slides contain one or more surface depressions ideal for liquid
solutions and larger specimens. These more expensive microscope slides can be used
without a cover.
Some manufacturers produce plastic chambers with a predetermined number of slides
with covers.
Filled calibrated wells or flasks are viewed quickly without preparing or clipping
individual slides to the microscope stage, making this especially useful in sediment
studies, such as urine analysis; in addition, some tray designs can be placed in an
incubator or refrigerator, allowing for the study of cultured samples.
Additional Features
Some cells and tissues cannot adhere to a plain glass surface and require a positive charge or surface
modifications.
Saving time and money, electrostatic charged slides are a popular choice for
researchers of histology, cytology and pathology.
Surfaces treated with biological reagents can make a slide water-proof, resistant to
certain chemicals and reduce instances of cross-contamination.
Additional variations to microscope slides include:
Etched grid system or graticule
Enables researcher to monitor and communicate area(s) of interest
Aids in hand sketching
Helps geographical plotting
Estimates size and scale
Dual concavity
Side-by-side comparisons, including sample to control
Reduces risk of cross-contamination
Transparent Mica
Rarely used substitute for glass
Less prone to dust, scratching
Prevents glare
Cover Slips
Almost always made of borosilicate or silicate glass, cover slips hold samples in place
and protect them from inadvertent movement and contamination.
It also protects the microscope, preventing direct contact between the sample and lens
as well as accidental leakage of water-based preparations.
The thin, transparent cover glass is usually square and available in types Number 1 and
Number 2.
Suited for high-resolution microscopy and oil immersion preparations, Number 1 covers
are .13-.17mm thick. Number 2 covers, .17-.25 mm, are designed for general purpose.
Less frequent characteristics include rectangular shapes, alternative materials such as
quartz and certain types of plastic, etched lines or grids and additional thicknesses.
If not creating a permanent slide with glue and/or sealant, cover slips can be removed
and sterilized for multiple reuses.
Preparation Techniques: Dry Mounts, Wet Mount,
Squash, Staining
The main methods of placing samples onto microscope slides are wet mount, dry
mount, smear, squash and staining.
Dry Mount:
The dry mount is the most basic technique: simply position a thinly sliced section on
the center of the slide and place a cover slip over the sample.
Dry mounts are ideal for observing hair, feathers, airborne particles such as pollens and
dust as well as dead matter such as insect and aphid legs or antennae. Opaque
specimens require very fine slices for adequate illumination.
Since they are used for primarily inorganic and dead matter, dry mounts can
theoretically last indefinitely.
Wet Mount:
Used for aquatic samples, living organisms and natural observations, wet mounts
suspend specimens in fluids such as water, brine, glycerin and immersion oil. A wet
mount requires a liquid, tweezers, pipette and paper towels.
To prepare the slide:
Place a drop of fluid in the center of the slide
Position sample on liquid, using tweezers
At an angle, place one side of the cover slip against the slide making contact
with outer edge of the liquid drop
Lower the cover slowly, avoiding air bubbles
Remove excess water with the paper towel
Although wet mounts can be used to prepare a significantly wide range of microscope
slides, they provide a transitory window as the liquid will dehydrate and living
specimens will die.
Organisms such as protozoa may only live 30 minutes under a wet mount slide;
applying petroleum jelly to the outer edges of the cover slip creates a seal that may
extend the life of the slide up to a few days.
In addition, larger protozoan such as paramecium may be too large and/or move too
quickly under the wet mount.
In these circumstances, adding ground pieces of cover glass to the water before the slip layer will create added
space and chemicals or strands of cotton can be added to slow the movement of paramecium, amoeba and
ciliates.
Smear Slides:
Smear slides require two or more flat, plain slides, cover slips, pipette and tissue paper:
Pipe a liquid sample such as blood or slime onto a slide
Using the edge of the second slide, slowly smear the sample creating a thin,
even coating
Put a cover slip over the sample, careful not to trap air bubbles
Remove excess liquid
Ideally, smears should dry naturally in an environment of moderate, steady
temperature.
The angle of the smearing slide determines the length of the smear; a steeper angle
creates a shorter smear. For samples such as blood, begin by backing the smearing
slide into the sample and then push across the slide, pulling the blood in the opposite
direction to create a smooth layer.
A thicker slide can be created with two drops, but only with the blood of mammals as
the erythrocytes lack a nucleus allowing cells to be amassed in multiple layers.
Read further about Blood Smears - process and technique, possible presence of
artifacts
Squash Slides:
Designed for soft samples, squash slides begin by preparing a wet mount; place lens
tissue over the cover glass; gently press down, careful not to destroy the sample or
break the cover glass, and squash the sample; remove excess water.
Staining:
A variety of methods exist for staining microscope slides, including non-vital or in vitro
stains of non-living cells and vital or in vivo stains of living tissue. Staining provides
contrast through color that reveals structural details undetected in other slide
preparations.
Staining solutions such as iodine, methylene blue and crystal violet can be added to wet
or dry mounts.
A simple staining method:
Add a drop of staining solution on the edge of one side of the cover slip
Position the edge of a paper towel on the opposite end
Allow dye to be pulled across the specimen
Stains are especially useful in the fields of histology, virology and pathology, allowing
researchers to study and diagnose diseases, identify gram positive and negative
bacteria as well as examine detailed attributes of a variety of cells.
How to study a microscope slide
Observing microorganisms under a microscope is a very simple and interesting activity! The procedure is
quite simple: The microscope slide should be placed on a microscope stage and then clamped with
special spring clips. But viewing slides under a microscope requires careful attention in order to increase
the educational effect and minimize the chance of breakage.
The detailed steps:
1. Put your microscope on a flat surface. If your microscope is equipped with an electric illuminator, make
sure that you have an outlet nearby.
2. To carry your microscope from one place to another, keep one hand on the tube holder, and the other
hand under the base of the tripod. This will allow you to avoid accidental drops and breakage of the
instrument.
3. If your microscope is equipped with an illuminator, connect the microscope to the power source to
turn it on, and then remove the eyepiece cover.
4. Holding the microscope slide by the edges, place it on the microscope stage (the cover slip should be
on top). Slowly move the microscope slide so the specimen is in the center right under the microscope
objective lens.
5. Turn the revolving nosepiece so that the objective lens with the smallest increase (usually the smallest
in size as well) is imposed on the specimen. Using the coarse adjustment knobs, lower the objective lens
as closely as possible to the slide, but without making contact with the slide. Control the process looking
directly at the microscope stage (not through the eyepiece). This is especially important when working
with high-powered objectives.
6. Looking through the eyepiece, slowly lift the objective lens away from the specimen, using the coarse
adjustment knobs, until the image is in focus. After that, use the fine adjustment mechanism to obtain
the maximum sharpness of the image.
7. Change the objective lens to a more powerful one by turning the revolving nosepiece. Remember to
control the distance between the objective lens and the stage to avoid damage to the objective or
specimen. Repeat step 5 - this time with a higher magnification power. Use this method every time you
change magnification of the instrument.
