Publication: Counting particles in tissue sections: Choices of methods and importance
of calibration to minimize biases
Authors
von Bartheld, C.S.
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Publisher
Murcia : F. Hernández
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DOI
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info:eu-repo/semantics/article
Description
Abstract
Investigators must choose between counting
methods to quantify microscopic particles in tissues. The
c o nventional profile-based ("model-based" or "2D-")
counting methods have been criticized for their potential
biases due to assumptions about shapes, sizes, and
orientation of particles when converting profile counts
into cell numbers. New stereological methods ("designbased"
or "3D-") methods such as the optical disector or
p hysical disector were initially introduced as being
inherently unbiased. Recent calibration analyses and
comparisons of results from different investigators have
r evealed the potential for significant biases in the most
efficient and most frequently used design-based method,
the optical disector. This rev i ew aims to objective l y
assess the strengths and limitations of current profi l e -
and disector-based cell counting methods by
examination of studies in which these methods have
been calibrated against the "gold-standard", counts
obtained by 3-dimensional reconstruction of serial
sections. Advantages and disadvantages of each counting
method and the associated embedding and sectioning
techniques are compared and frequent mistakes and
pitfalls of each technique are discussed. The importance
of a calibration step for each technique is emphasized,
and a protocol is provided for a quick and simple
calibration by a "sampling" 3-D reconstruction of limited
serial sections. Trends in the usage of counting methods
are analyzed in four major journals. It is hoped that this r ev i ew will be helpful, for both inv e s t i gators and
manuscript rev i ewers, in clarifying some of the
contentious issues in the choice and implementation of
appropriate methods for particle counting in tissue
sections.
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