Preparing Histopathology Slides

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Histopathology Slides The First Steps to Prepare
the Highest Quality Slides
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To analyze any specimen in your histology lab,
you must prepare a perfect histopathology slide
first. For this, you should fix the specimen in
such a way that light passes evenly through it
and demarcates all its significant
features. Thus, preparing highest quality slides
begins with the few first steps.
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• What are these first steps?
• Here they are
• Choose the highest quality glass for your slides
  and cover slips.
• Choose the slides that fit on your microscope
  equipment and fulfill the purpose of your
  examination.
• Use the right slide mount.
• Use an effective mounting medium.
• Choose the right slide stains
• Lets discuss these steps in detail

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The glass slide

• The first step to prepare the highest quality
  slide is to choose the best kind of slide glass.
  Some glass slides have floating impurities in
  them which may create patches of false color and
  reduce the contrast and clarity.
• Microscope slides are generally made of optical
  quality glass. Some are made of soda-lime or
  borosilicate glass or special transparent
  plastics of optical quality. Choose one that fits
  your purpose of the experiment.
• If you will use fluorescent light to illuminate
  the specimen, use slides made from fused quartz.

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• The cover slips should be made of the highest
  quality optical glass and must be as thin as
  possible to reduce film effects.
• Glass slides are also made in various shapes for
  different purposes. For example, concave slides
  have a shallow upper surface to hold liquids.
  Graticule slides have measurement grids on
  their surface to aid in counting the cells or
  assessing the size of the objects. Some slides
  have special reservoirs and channels cut into
  them.

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The mount

• Mounting is the process of putting the specimen
  on the slide. The way the subject is mounted on
  the slide can affect the clarity of its image.
  The various slide mounts are
• Dry mounting places the thin, light,
  transmittable specimen onto the slide. A
  coverslip is placed on the top of the specimen
  to keep it flat and avoid contamination from the
  microscopes lens.

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• In a wet or temporary mount, the specimen is
  immersed in water or another liquid on the slide.
   A coverslip is placed to flatten the specimen
  and avoid refraction effects from the liquid.
• A permanent mount is used for animal or human
  tissue studies. All water is removed from the
  cells of the specimen which is then sliced with
  the microtome and placed on the slide. A stain is
  applied to bring out its important features. A
  coverslip is placed to stabilize and protect the
  slide.

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Mounting media

• Wet mounts use water or glycerol as the mounting
  media.
• Permanent mounting uses substances which can
  thicken and hold the specimens in place for a
  long time, thus, reducing decay and
  contamination.
• The mounting medium should not act as a lens
  otherwise, it will add additional light
  refraction and distortion. Plus, the mounting
  medium should have the same refractive index as
  the specimen.

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The stains

• Stains help to highlight the particular features
  of the cell anatomy.  Varieties of stains are
  used in a histopathology laboratory depending
  upon the specific purpose.
• The quality of your slide also depends on the
  appropriate use of stains. So, choose wisely
  according to what you wish to accomplish through
  your microscopy work.

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Taking care of these first steps will help you
prepare the highest quality histopathology
slides, save your time and money, and give you
your desired results.
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