Abstract
Aims-To investigate (1) whether adequate
immunohistochemical staining can
be achieved on sections cut from plastic
embedded bone marrow trephine biopsy
specimens after microwave heating in citrate
buffer; and (2) whether this immunohistochemical
staining is comparable with
that achieved on routine sections cut from
paraffin wax embedded trephine biopsy
specimens after decalcification procedures.
Methods-Sixty five consecutive bone
marrow trephine biopsy specimens of
more than 1 cm in length were divided
transversely into two equal parts. One part
was processed in paraffin wax foliowed by
decalcification. The other part was embedded
in the epoxyresin Polarbed 812 followed
by the cutting of 1 pm sections. Both
parts underwent immunohistochemical
staining by an identical panel of antibodies.
With Polarbed 812 plastic embedded
sections, microwave heating in
citrate buffer was undertaken before the
application of antisera.
Results-On sections cut from plastic embedded
material, immunohistochemical
staining was generally satisfactory, easy
to interpret and comparable with that
achieved with paraffin wax embedded
material. Exceptions were antibodies
to neutrophil elastase and CD61 where
immunostaining was consistently negative
on plastic embedded sections. Immunohistochemical
staining for CD20 was
consistently more reliable on plastic
embedded sections.
Conclusions-The results provide evidence
that, with few exceptions, satisfactory
immunohistochemical staining is possible
on plastic embedded bone marrow trephine
biopsy specimens after microwave
heating in citrate buffer. This, combined
with the advantage of superior cellular
morphology with semi-thin (1 pm) sections
of plastic embedded material, make
such embedding procedures the preferred
method for the processing ofbone marrow
trephine biopsy specimens.
ABSTRACT
The microwave has always promised to shorten tissue processing times. However, historically much of the use of the microwave has produced inadequate and varied results. This science has been further
hampered by assumptions and misunderstandings as to how the microwave shortens processing times. This article discusses the
important variables and advances in microwave processing and also illustrates why it was not technically possible for the variables to have been discovered earlier. Examples of protocols which can be used to standardize microwave processing techniques to produce consistent high quality results are given. The new methodology is
contrasted with the methodology of the past 30 years to illustrate the new advances in microwave tissue processing.
KEYWORDS:
transmission electron microscopy, light microscopy, microwave processing, fixation, paraffin processing, immunolabeling, invivo
labeling