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Histochemistryasatoolinmorphological
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Annals of Diagnostic Pathology 16 (2012) 71 – 78
History of Pathology
Histochemistry as a tool in morphological analysis: a historical review
Mark R. Wick, MD⁎
Divisions of Surgical Pathology & Cytopathology and Autopsy Pathology, University of Virginia Medical Center, Charlottesville, VA, USA
Abstract
Keywords:
Histochemistry has an interesting history, extending back to ancient times, in some ways. Man has
long had a desire to understand the workings of the human body and the roles that various “humors”
or chemicals have in those processes. This review traces the evolution of histochemistry as an
investigative and diagnostic discipline, beginning with the efforts of medicinal chemists and
extending through a period in which histology was increasingly paired with biochemistry. Those
developments served as the underpinnings for an eventual marriage of microscopy, chemistry,
immunology, and molecular biology, as realized in the current practice of anatomical pathology.
© 2012 Elsevier Inc. All rights reserved.
Histochemistry; Histology; Immunohistochemistry; Biochemistry
One can defensibly argue that biochemistry and histology
originated from the same human interest, that is, a desire to
know the basic structure and composition of living things.
From the beginning of time, a series of observations—both
scientific and fanciful—accrued in an effort to inform that
topic. Such “data” emanated from several and diverse
sources, such as hunter-gatherers, alchemists, mathematicians, abbatoir workers, physicians, anatomists, morticians,
astrologers, sorcerers, philosophers, and theologians [1-4].
In ancient Greece, Hippocrates theorized that diseases
were caused by imbalances in 4 basic body substances, called
humors: phlegm, blood, black bile, and yellow bile [4,5].
Astoundingly, variations on that mechanistic scheme were
accepted as dogma until the 19th century. Diets designed to
“cleanse putrefied juices” were therapeutically joined with
purging or venesection or both to reestablish a balance
between the 4 humors [6]. Theophrastus Phillippus Aureolus
Bombastus von Hohenheim (1493-1541; also known as
Paracelsus) was among the first to challenge such views and
practices [4]. He believed that illness was induced by factors
originating without, rather than within, the body, and that it
resulted—at least partly—from imbalances of indigenous
chemicals and minerals [7]. As a corollary to that premise,
Paracelsus encouraged investigations of the compounds and
⁎ University of Virginia Hospital, Charlottesville, VA 22908-0214,
USA. Tel.: +1 434 242 2410.
E-mail address: [email protected].
1092-9134/$ – see front matter © 2012 Elsevier Inc. All rights reserved.
doi:10.1016/j.anndiagpath.2011.10.010
elements that comprised plant and animal tissues. Moreover,
he opened the door to the bona fide practice of pharmacy, in
which prescribed external substances were taken into the
body and targeted to the presumed sources of biochemical
aberration or deficiency [4].
That approach clearly affected the primary focus of
medicine in the middle ages, which was nonmorphological
and primitively centered on biologic chemistry. Physiologic
mechanisms and anatomical structure were regarded as
relatively inconsequential during that period of history.
Therefore, no disadvantage was attached to destructive
(digestive) analysis of plants and animals, in efforts to discern
their chemical constitution [8].
Beginning in the 16th century and through the efforts of
Andreas Vesalius, William Harvey, Anton van Leeuwenhoek,
and others, the study of anatomical structures grew steadily at
gross and microscopic levels [1,3]. Botany was the principal
scientific discipline in which such activities evolved; early
textbooks on the subject of plant histochemistry included
Essai de Chimie Microscopique Appliquee a la Physiologie
and Nouveau Systeme de Chimie Organique, both by
Francois-Vincent Raspail (Fig. 1) (1830 and 1833, respectively) [9,10]; Lehrbuch der physiologischen Chemie by Karl
Gotthelf Lehmann (1842) [11]; and Handbuch der Experimental Physiologie der Pflanzen by Julius von Sachs (1865)
[12]. Interestingly, botanists retained a basic interest in the
cellular chemical processes that were illumined by histochemistry, whereas zoology-oriented histologists and histochemists used microscopy and staining techniques primarily
72
M.R. Wick / Annals of Diagnostic Pathology 16 (2012) 71–78
a science can be traced to the introduction by Bencke, in
1852, of the aniline dyes which were in general use by 1880,
followed closely by the development of paraffin-sectioning
and photomicrography as routine techniques. Thus, by the
end of the [19th] century, a fashion was set in histology
which even today has not been completely supplanted. In
retrospect, it may seem strange that the attention of
histologists was concentrated for so long upon descriptive
morphology without their making any serious attempt to
study the chemistry of the structures they were staining… it is
the use of a staining procedure with known chemical
specificity that distinguishes a histochemical from a
histological technique” [15].
The foregoing material sets the stage for a discussion of
the 3 main categories into which “histochemists” can be
assigned over the past 150 years. These are (1) investigators
who were chemistry-oriented but not concerned with
morphology; (2) those with contemporaneous interests in
physiologic chemistry, histology, and technology; and (3)
applied (diagnostic) histochemists (histopathologists).
1. “Histochemists” who were indifferent to morphology
Fig. 1. Etching of Francois-Vincent Raspail (1794-1878), one of the
originators of histochemistry as a discipline. Raspail was also a naturalist
and a politician.
to further the development of microanatomy, taxonomy, and
nosology, more or less in vacuo.
The latter situation led to an interesting, Darwinesque
competition. Physiologic-cellular chemists began to disparage the efforts of histologists in the second half of the 19th
century as unworthy of true scientific respect. Morphologists
were regarded as little more than clerks and scribes who
recorded their visual observations without correlating them
to chemical findings [13]. That perception was furthered by
the tendency of many histologists to embrace new stains and
dyes as a means to an end (ie, morphological discrimination),
rather than as ligands for cellular chemicals that had yet to be
delineated. Pearse [8] framed this picture well, in saying,
“although diagnostic significance was attached to many of
the new color reactions, no attempt was made to put them on
a physical or chemical basis.” Hence, in the era introduced
by August Bencke in the 1860s and 1870s, with aniline dyes
and similar reagents in hand, histology-based histochemists
broke ranks, in philosophical and heuristic terms, with
cellular biochemists [14]. A rancorous dichotomy persisted
between the 2 groups well into the 20th century; indeed, as
late as 1962, Lewis [15] stated that “the decay of histology as
As mentioned earlier, one, rather extreme, view of living
organisms was that their structure is only important as a way
of partitioning inorganic and organic substances, or chemical
reactions. This was the credo of “pure” biochemists, who
typically subscribed to “destructive” or “digestive” histochemistry. In such a context, the possible affinity of tissue
for chemical laboratory reagents had “meaning” only if it
illuminated the biochemical constitution of the substrate [8].
An example is represented in an early analysis, by FrancoisVincent Raspail, of starch in plant tissues, using the binding
of iodine solution as an indicator [9]. The amylose in plant
carbohydrates enters a colloidal suspension in water and
comprises long polymeric chains of glucose units that are
interconnected by alpha-acetal linkages, forming a 3dimensional coil. Iodine molecules can intercalate with the
amylose coil, yielding a blue moiety [10,16]. The latter
property obtains, regardless of whether the target is groundup plant material studied in a test tube, intact amylose-rich
organs that are infused with potassium iodide (Fig. 2) or
histologic sections of tissue that are “stained” with iodine
and visualized with a microscope. To histochemists
belonging to the pragmatic group under discussion, it would
not matter—it would be sufficient to know that the target
tissue did indeed contain starch, explaining its iodinophilia.
Similar comments can be made regarding iron deposits
(hemosiderin) in plants and animals. Perls [17] was among
the first to show that acidified potassium ferrocyanide
solution binds to iron in tissue, forming a relatively insoluble
blue-purple precipitate with the chemical formula Fe7(CN)18(H2O)x, where 14 ≤ × ≤ 16. Again, in an egalitarian
sense, it might be regarded as immaterial whether the iron
was demonstrated in a glass beaker, an intact organism, or a
M.R. Wick / Annals of Diagnostic Pathology 16 (2012) 71–78
Fig. 2. Cut section of amyloidaceous myocardium, stained with
Lugol's solution (elemental iodine and potassium iodide in solution).
Because amyloid contains amylose-like carbohydrate sequences, the heart is
stained blue.
microscope slide. Analogous models include the demonstrations of peroxidase in pus by Klebs [18] using tincture of
guaiac in 1868, Ehrlich's [19] detection of cytochrome
oxidase (originally called “Nadi oxidase”) in 1885 by
intravenous injection of alpha-naphthol and p-phenylenediamine into animals, Rudolf Heidenhain's discovery that
selected cells in the gastric mucosa would turn brown when
exposed to chromic acid (“chromaffinity”) [20], and
Miescher's [21] identification of DNA in cellular nuclei
through its selective binding to methyl green.
All of these assessments provided new information, but,
from the perspective of current-day morphologists, they
would be unsatisfying because the particular cellular
locations of the chemical substances in question were not
addressed. In that vein, microanatomy as a discipline was
advancing in the 1800s as well, despite its being regarded as
a “non-science” by biochemists of the period. The first
attempt at a comprehensive textbook of histology was
published in 1841 by Friedrich Gustav Henle [22], followed
by a succession of additional works by Albert Donne, Arthur
Hill Hassall, Rudolph Albert von Kolliker, Lionel Beale,
Gottlieb Gluge, John Scott Burdon-Sanderson, Georg
Eduard von Rindfleisch, Andre-Victor Cornil and LouisAntoine Ranvier, Edward Albert Schafer, Phillip Stohr, and
other authors in the latter half of the 19th century [23]. The
topical approach in several of those publications was to
combine microanatomy with physiology, stressing both
structure and function simultaneously. That orientation led to
development of the next tier of histochemists, whose work
occupied much of the 20th century.
73
enthusiastically. Nevertheless, as cited earlier, that practice
was often unaccompanied by a clear understanding of exactly
what was being labeled by tissue staining procedures.
Moreover, the detailed chemical mechanisms for such
techniques commonly went unstudied as well. Dyes that had
entered into biologic use included “cochineal” agents such as
mucicarmine [24], aniline dyes [25], hematoxylin and its
congeners [26], precipitable silver solutions [27], Schiff-base
derivatives [28], colloidal suspensions of metal ions [29],
phthalocyanines [30], cotton dyes (eg, Congo red, Pagoda red)
[31], methyl and ethyl green [32], and others.
An international group of investigators increasingly focused
on the biochemical processes and targets that were associated
with the use of such reagents, in the 1890s and beyond. They
included—but were not limited to—individuals such as Paul
Ehrlich, Santiago Ramon y Cajal, Karl Weigert, Pio del RioHortega, Joseph von Gerlach, Paul Mayer, Friedrich Miescher,
Alfred Fischer, Gustav Mann, Robert Feulgen, Julius von
Kossa, Frank Burr Mallory, Lucien Lison, Clyde Mason,
Maffo Vialli, Emile Chamot, David Glick, George Gomori,
Anthony Guy Everson Pearse, and Ralph Lillie [8]. Some early
explanations for the cellular affinities of dyes were scientifically infantile, for example, Fischer suggested in 1899 that all
stains were merely absorbed passively by tissue [33].
Conversely, Ehrlich [34] and Miescher [21] correctly believed
that specific chemical coupling was responsible, and in his text
titled Physiological Histology—published in 1902—Mann
2. “In situ” biochemical histochemists
Given the availability of dyes that evolved during the mid
1800s, histologists of the period began to use them
Fig. 3. Anthony Guy Everson Pearse, MA, MD, FRCPath, DCP, FRCP
(1916-2003), Professor of Histochemistry at the Royal Postgraduate Medical
School in London, England.
74
M.R. Wick / Annals of Diagnostic Pathology 16 (2012) 71–78
Table 1
Histochemical methods now in use in anatomical pathology
Histochemical stain
Principal use
Secondary use
Acid-fast bacilli (Ziehl-Neelsen) stain
Identification of mycobacteria and selected other
microorganisms in granulomatous diseases
Identification of acidic and neutral mucins in
human cells and microorganisms
Identification of neuronal axons
Identification of neuronal axons
Identification of bacteria and selected other
microorganisms
Labeling of acidic and neutral mucins in human
cells and microorganisms
Identification of amyloid
Labeling of tissue copper deposits
Labeling elastic tissue
Identifying fibrin deposits
Labeling nontuberculous mycobacteria and
selected other microorganisms in tissue
Identification of melanin
Identification of mast cell granules and lipofuscin
Alcian blue stain
Bielschowsky silver technique
Bodian silver technique
Brown and Brenn (tissue Gram) stain
Colloidal iron stain
Congo red stain
Copper stains (rhodanine and orcein)
Elastic (Verhoeff-van Gieson) stain
Fibrin (Fraser-Lendrum) stain
Fite's acid-fast stain
Fontana-Masson stain
Giemsa stain
Gridley silver method
Grimelius silver stain
Grocott methanamine-silver method
Hall stain
Iron (Perls; Prussian blue) stain
Jones silver stain
Leder stain
Luxol fast blue
Masson trichrome stain
Methyl green–pyronin stain (MGP)
Mucicarmine (Mayer) stain
Nissl substance (cresyl violet) stain
Oil-Red-O stain
Periodic acid Schiff method
Phosphotungstic acid–hematoxylin stain
Reticulin (Sweet) method
Steiner technique
Toluidine blue stain
Trichrome (Masson) stain
Urate (DeGalantha) stain
Von Kossa stain
Weigert stain
Identification of primary granules in myeloid and
mast cells
Identification of amoebae and fungi in tissue
Identification of neurosecretory granules in
neuroendocrine tumors
Labeling of fungi (including Pneumocystis)
Identification of bile pigment
Identification of hemosiderin pigment
Labeling of basement membranes
Identification of myeloid-cell and mast-cell granules
Labeling of myelin in central and peripheral
nervous tissue
Differentiation of collagen, elastic tissue, muscle,
and epithelium
Labeling of nucleic acids (DNA and RNA)
Labeling of neutral (epithelial) mucins in tissues
Identification of extranuclear ribonucleic acid in
neurons and other cell types
Identification of lipid deposits (requires use of
frozen tissue)
Identification of glycogen (undigested tissue) or
neutral mucins and basement membrane material
(after tissue digestion with diastase)
Labeling of myofilaments, especially in striated
muscle cells
Identification of reticulin fibers (type III collagen)
in connective tissue
Silver-impregnation method for identification of
spirochetes and Legionella bacteria in tissue
Identification of myeloid and mastocytic granules
in tissue sections or blood smears
Differential staining of collagen (blue), muscle
(red), and elastic tissue (purple) in tissue
Labeling of urate deposits in tissue
(best used with alcohol fixation)
Identification of calcium salts in tissue
Labeling of myelin in neural tissue
stated that “it is not sufficient to content ourselves with using
acid and basic dyes and speculating on the basic or acid nature
of the tissues or to apply color radicals with oxidizing or
reducing properties… we must endeavor to find staining
reactions which will indicate not only the presence of certain
None
Labeling of reticulin and mucosubstances
Labeling of reticulin and mucosubstances
Labeling of high-molecular-weight keratin
None
Labeling of foreign material containing cellulose
Identification of hepatitis B virus in hepatocytes (orcein)
Labeling collagen and enhancing nuclear detail
Labeling collagen and high-molecular-weight keratin
Labeling mastocytic granules and lipofuscin
Labeling neurosecretory granules in argentaffin cells;
labeling neuromelanin
Labeling of protozoan microorganisms; identification of
amyloid (with metachromasia)
Labeling of reticulin fibers and mucins
Labeling of mucins
Labeling of mucins
None
None
Labeling of mucins
None
Identification of neurolipofuscin
Identification of selected protozoa
None
Identification of selected microbes
Metachromatic labeling of amyloid
Labeling of lipochrome
Labeling of selected microorganisms, especially fungi
Labeling of fibrin deposits
Labeling of mucins in tissue
Labeling of mucins in tissue
Labeling of protozoan organisms in tissue; metachromatic
staining of amyloid
Identification of selected microbes (eg, amoeba; helminths)
None
None
None
elements such as iron or phosphorus, but the presence of
organic complexes such as the carbohydrate groups, the
nucleins, protamines, and others” [35].
Most scientists in the above-listed group took that
directive to heart, as did others after them. Indeed, many
M.R. Wick / Annals of Diagnostic Pathology 16 (2012) 71–78
Table 2
Histochemical methods: undifferentiated large-cell neoplasms
75
Table 4
Histochemical diagnosis of small round-cell tumors
Tumor
PAS
Mucin
MGP
FM
Retic
Tumor
PAS w/o
Pericellular reticulin
Carcinoma
Germ-cell tumor
Lymphoma
Melanoma
MESO
+/−
+
0
+/−
+/−
+/−
0
0
0
0
+/−
+/−
++
+/−
+/−
0
0
0
+/−
0
+−OP
+−OP
+−PCP
+−OP
+−OP
PNET
RMS
Lymphoma
Neuroblastoma
+ to +++
+ to +++
0
0
0
+
+ to ++
0
PAS indicates periodic acid–Schiff; MGP, methyl green–pyronin; FM,
Fontana-Masson; Retic, reticulin; OP, organoid pattern; PCP, pericellular
pattern; MESO, epithelioid mesothelioma.
microscopists became so engrossed by a characterization of
in situ chemical reactions that the practical and diagnostic
uses of histochemistry were given short shrift. The
admonitions of Mann [35], and Lewis after him [15],
became the marching orders of the day. Histochemical
textbooks written by Lison [36] in 1936, Glick [37] in 1949,
Gomori [38] in 1952, Pearse [39] in 1953, Lillie [40] in
1954, Bancroft [41] in 1967, Kiernan [42] in 1981, and
Sumner [43] in 1988 were devoted largely to the chemistry
of tissues as seen under the microscope. As a result,
knowledge of cellular biochemistry grew exponentially
during the 20th century. By 2000, Coleman [44] was able
to say confidently that “histochemistry and cytochemistry…
allow precise analysis of the chemistry of cells and tissues in
relation to structural organization.” He also went on to state
that histochemistry was still a useful and productive field of
study and that “there are…few other disciplines in experimental biology or medicine that can make a similar claim.”
3. Histochemists with a diagnostic orientation
As mentioned earlier in this discussion, philosophical
tension has existed between “basic” and “applied” histochemists for well over 100 years. This is not a novel situation, and
in fact, it has applied to every one of the “translational”
scientific techniques used in morphology-oriented areas of
laboratory medicine. Electron microscopy, immunohistology, in situ hybridization, polymerase chain reaction–based
procedures, and other “blotting” technologies have served as
comparable battlegrounds for purists and practitioners [45].
In 1955, Jonas Friedenwald—a “basic” researcher in
ophthalmology at Johns Hopkins University—published a
review of applied histochemistry, including in it several
Table 3
Histochemical methods in selected dermatological conditions
• Lupus erythematosus (stromal mucin stains; PAS-D to show thick EBM)
• Granuloma annulare (stromal mucin stains show increased interstitial mucin)
• Porphyria cutanea tarda (PAS-D stain shows EBM abnormalities)
• Perforating dermatoses (trichrome and VVG stains demonstrate extrusion
of dermal connective tissue through epidermis)
Abbreviations: PAS-D, perioric acid Schiff stain with diastase digestion;
EBM, epidermal basement membrane; VVG, Verhoeff-van Gieson stain.
PAS w/o means periodic acid–Schiff stain without diastase.
PNET indicates primitive neuroectodermal tumor; RMS, rhabdomyosarcoma.
maxims that are still true [46]. In regard to criticisms that
focused on the “nonspecificity” or crudeness of some
histochemical reactions, he said “criteria of specificity [in
histochemistry] are similar to those in qualitative chemistry in
general… [they] can be very much enhanced if two or more
different reactions can be applied and compared.” Presciently, he went to opine that “analysis in-situ is a non-quantitative
procedure. Sensitivity, therefore, merely concerns the limits
at which the reaction is discernible.” The latter comments
apply equally well today, in reference to modern attempts at
“quantitative” immunohistochemistry [47].
Three years earlier, Robert Stowell—chair of pathology
at the University of California-Davis—had suggested that
“fundamental, critical research on new cytochemical techniques will do more to advance our eventual understanding
of normal tissues and neoplasia than the application of the
relatively few and often none-too-satisfactory histochemical
and cytochemical techniques now available” [48]. In
counterpoint, Pearse—arguably the most well-versed histochemist of all (Fig. 3)—responded thus to Dr Stowell: “in
medicine the new and imperfect remedy does not await
perfection by the research of groups of collaborating
investigators in the pure sciences. It is applied forthwith
to…patients by the practitioners of medicine, and it is often
by their observations and research that real advancement in
the use of the remedy, and in knowledge of its mechanism
and meaning, is brought about. I believe very strongly,
therefore, that the methods of modern histochemistry,
despite their imperfections, should be applied by all
practitioners in the biological, cytological, and pathological
sciences” [49]. After the passage of another 20 years, Pearse
could further state that “histopathology can be transformed
by the application of any technology which confers upon its
observations an increase in objectivity. Foremost in the field
comes histochemistry, for a variety of reasons. These include
sheer breadth of scope and overwhelming numerical
superiority in respect of techniques” [50].
Pearse [39], Bancroft and Stevens [51], and Filipe and
Lake [52] took such tenets and built textbooks around them
in the latter part of the twentieth century. With such
perspectives by experienced hospital pathologists, the place
of histochemistry as a valuable clinical method was
solidified. Despite refinement and flux in the nosologic
categorization of some human diseases, histochemical
analysis continues to offer important information in regard
to histopathologic diagnosis and differential diagnosis. A
76
M.R. Wick / Annals of Diagnostic Pathology 16 (2012) 71–78
Fig. 4. A, This tumor of the lung is labeled with the Best mucicarmine method, demonstrating the presence of abundant intracellular epithelial mucin and
supporting a diagnosis of adenocarcinoma (mucicarmine, ×300). B, The liver in sickle-cell disease shows easily seen hemosiderin deposits, using Perls' method
(Perls stain, ×200). C, This epithelioid neoplasm of the dermis exhibits diffuse reactivity with the chloroacetate estrase (Leder) stain, consonant with its identity
as a granulocytic sarcoma (Leder stain, ×200). D, chromaffinity is observed in this ileal carcinoid tumor, using the Fontana-Masson technique (Fontana-Masson
stain, ×200).
sampling of histochemical methods now in use in anatomical
pathology is presented in Table 1. Tables 2 to 4 and Fig. 4
show selected practical applications of selected stains in
Table 5
Selected infectious organisms requiring special histochemical techniques for
identification
• Spirochetes, Legionella, Bartonella, and Yersinia—require use of the
Dieterle or Warthin-Starry silver stains and will not label with BrownHopps method
• “Atypical” mycobacteria—best recognized with the Fite procedure
• Rhodococcus, Legionella micdadeii, and Nocardia are also acid fast with
the Ziehl-Neelsen procedure
• Dematiaceous fungi (as in chromoblastomycosis and phaeohyphomycosis)
are Fontana-Masson positive because of melanin content
• Viruses can be labeled with the methyl green–pyronin e stain, as well as
Macchiavello method and Lendrum technique
well-defined histologic contexts. Table 5 shows selected
infectious organisms requiring special histochemical techniques for identification.
4. The nexus of histochemistry with immunology and
molecular biology
Dr Albert Coons was still a house-officer at Massachusetts General Hospital when he conceived a simple but
revolutionary idea. His thought was to label antibodies with a
chemical tag, so that their binding to predefined antigens in
tissue could be visualized microscopically. Despite the fact
that antibody structure was only primitively understood at
that time and the lack of a proven technique for artificially
M.R. Wick / Annals of Diagnostic Pathology 16 (2012) 71–78
77
However, in place of antibody probes to which chemical
indicators can be joined, the basic investigative tools in in
situ hybridization are specific sequences of nucleic acid that
are complementary to DNA or RNA targets of interest in
tissue sections [57,58] (Fig. 6).
5. Conclusions
Fig. 5. Direct immunofluorescence microscopy of a skin biopsy in bullous
pemphigoid, which was labeled with fluorescein-tagged antibody to
immunoglobulin G (EgG). Linear reactivity is seen at the epidermal
basement membrand (Anti-IgG immunofluorescence, ×200).
binding other substances to them, Coons pursued the concept
doggedly over several years [53]. Eventually, specific
antibodies were produced in vivo in animal hosts that were
specific for particular proteins. They were coupled successfully with fluorescein isocyanate and proved to be effective
in localizing polypeptide targets in histologic sections that
were illuminated by ultraviolet light with a special
microscope [54,55] (Fig. 5). Hence, the field of immunofluorescence-based histochemistry was thereby established by
Coons, who won the prestigious Albert Lasker award in
1959 for that contribution [56].
Today, many different chemical tags can be linked with a
plethora of antibody reagents that are clinically relevant in
pathology. In addition, the facet of molecular biology known
as in situ hybridization is predicated on a similar construct.
Fig. 6. Chromagenic in situ hybridization (CISH) of a uterine cervical
squamous carcinoma for human papillomavirus type 16 DNA. Several gene
copies are seen in each tumor cell nucleus (CISH, ×250).
Histochemistry has had a long history as well as a broad
interface with many of the other life sciences. Because the in
situ chemical reactions it concerns have been thoroughly
studied over many years, histochemistry is now one of the
most objective methods in biology and medicine. That fact
should not be forgotten in the current fervor over “new”
techniques in pathology, nor should one slight the practical
use of histochemistry in a variety of clinical differential
diagnostic settings. The rapidity, reproducibility, and
relatively low expense attached to this form of biomedical
analysis continue to recommend it as a valuable enterprise,
after nearly 200 years of existence.
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