Evolution of Histochemistry: A Journey Through Scientific Breakthroughs
Lecture 1:
Lecture 1:
Historical Review on
Historical Review on
Histochemistry and
Histochemistry and
Classification of
Classification of
Histochemical Methods
Histochemical Methods
Course Code:
Course Code:
ANA 304
ANA 304
Course Title:
Course Title:
General Histochemistry
General Histochemistry
Course Lecturer:
Course Lecturer:
Edem E. Edem
Edem E. Edem
Course Outline
LECTURE ONE:
•
1.1
Historical review on histochemistry and cytochemistry
•
1.2
Classification of histochemical methods
•
1.2.1
Chemical methods
•
1.2.2
Physical methods
•
1.2.3
Biological methods
LECTURE TWO:
•
2
Histological and Histochemical Techniques
•
2.1
Animal treatment and tissue processing
•
2.2
Fixation of tissues and cells
•
2.2.1
Chemical fixation for insoluble compounds
•
2.2.2
Chemical fixation for soluble compounds
•
2.2.3
Cryo-fixation for soluble compounds
•
2.3
Embedding and sectioning
•
2.3.1
Cryo-sectioning without embedding
•
2.3.2
Freeze-drying and embedding
•
2.3.3
Freeze-substitution and embedding
•
2.3.4
Dry sectioning of freeze-dried or freeze-substituted materials
•
2.4
Observation of histochemical specimens by microscopy
•
2.4.1
Light microscopy
•
2.4.2
Electron microscopy
•
2.4.3
Image analysis of histochemical reactions
Edem, E. E.
2
Course Outline
LECTURE THREE:
•
3.
Histochemical Methodology - General Histochemistry
•
3.1.
Chemical methods in histochemistry
•
3.1.1.
Color reactions for light microscopy
•
3.1.2.
Electron dense deposits for electron microscopy
•
3.2.
Physical methods in histochemistry
•
3.2.1.
Methods using high or low temperatures
•
3.2.2.
Methods using lights with different wave lengths
•
3.2.3.
Methods using radiations
LECTURE FOUR:
•
3.3.
Biological methods in histochemistry
•
3.3.1.
Immunohistochemistry
•
3.3.2.
Lectin histochemistry
Edem, E. E.
3
Course Outline
LECTURE FIVE:
•
4.
Special Histochemistry - Histochemistry of the Organs
•
4.1.
The skeletal system
•
4.2.
The muscular system
•
4.3.
The circulatory system
•
4.4.
The digestive system
LECTURE SIX:
•
4.5.
The respiratory system
•
4.6.
The urinary system
•
4.7.
The reproductive system
•
4.8.
The endocrine system
LECTURE SEVEN:
•
4.9.
The nervous system
•
4.10.
The sensory system
•
5.
Revision
Edem, E. E.
4
Objectives
At the end of the class, students should be able to:
•
Define histochemistry
•
Elucidate the goals of histochemistry
•
Appreciate the historical development of
histochemistry as a scientific discipline
•
Identify the related disciplines to histochemistry
•
Classify the different methods used in
histochemistry
Edem, E. E.
5
What is Histochemistry?
Edem, E. E.
6
•
Histo
= histology:
chemistry
= chemical
reactions
•
The branch of science concerned with the
qualitative and quantitative assessment of
chemical compounds in a cell/tissue using
stains/dyes and microscopy.
•
It is a marriage between bio/chemistry and
cyto/histology
Historical Review on Histochemistry
Edem, E. E.
7
•
Histochemistry/cytochemistry is as old as histology
itself
•
In early 19th century, histochemical researches to study
chemical components of biological structure in
combination with chemistry and biology started first in
botany in France (Raspail,1825).
•
He stained starch in plant tissues blue with potassium
iodide solution under the light microscope and
demonstrated its localization microscopically
•
Then, he published an essay on microscopic chemistry
for the first time (Raspail, 1825)
Historical Review on Histochemistry
Edem, E. E.
8
•
Later, from the 1840’s to 1870’s, histochemistry in
zoology and medicine was developed mainly as
biological chemistry, together with histology
(Lehmann,1842).
•
Among these zoologists, anatomists and pathologists
who were at that time interested in analyzing chemical
constituents in animal tissues including human, Miescher
(1874) was the first to introduce cell fractionation to
analyze nucleic acids in nuclei of leukocytes.
•
During these times in the 19th century, this new field
was called as “microchemie” in French or
“Mikrochemie” in German, which meant
microchemistry in English.
Historical Review on Histochemistry
Edem, E. E.
9
•
Microchemistry was, in other words,
microscopic chemistry or chemical microscopy
and meant to observe chemical reactions
in situ
under microscopy.
•
In early 20th century, aniline dyes were
frequently used to stain tissues in anatomy and
pathology.
•
Histologists and pathologists were much
interested in new dyes and less interested in
histochemistry at that time.
Historical Review on Histochemistry
Edem, E. E.
10
•
When Lison (1936) published his famous book
entitled
“Histochemie Animale”,
many histologists
were again interested in histochemistry.
•
Lison classified histochemical techniques into
two categories,
“méthodes extra situm”
or
“méthodes extractives”
and
“méthodes
histochimiques in situ”
or
“méthodes
topochimique”
in French.
•
He proclaimed Histochemistry to be the
new
science
Historical Review on Histochemistry
Edem, E. E.
11
•
Then, many histologists, anatomists, pathologists,
physiologists started to study this new field and
published many original papers and books dealing
with histochemistry and cytochemistry.
•
Innumerable literature, as well as textbooks and
handbooks, are available from the 1950’s to 1990’s.
•
Nagata (1995) proposed
that the field should be
designated General Histo-cytochemistry just like we
have General Histology.
•
One of the foremost histochemists in Nigeria is the
Late Prof. Caxton-Martins
Classification of Histochemical Methods
Edem, E. E.
12
•
Nagata (1995) proposed the classification of
histochemical methods into:
1.
Chemical methods such as chemical
reactions by staining
Light microscopic photograph of
the colon of an adult mouse
stained with HID-AB sequence.
The goblet cells in
the top region (upper half of the
picture) of the crypts stained
black with HID (high iron
diamine), while the goblet cells in
the deeper region (bottom)
stained blue with AB (Alcian
blue). x480.
Classification of Histochemical Methods
Edem, E. E.
13
ii.
Physical methods such as radiations
Light microscopic radioautogram of the colon of a mouse injected with 35SO
4
,
fixed at 180 min., sectioned, radioautographed and stained with toluidine blue.
Classification of Histochemical Methods
Edem, E. E.
14
iii.
Biological methods such as immunity
Light micrograph of an adult mouse kidney cortex, cryosectioned, stained with biotinylated Lotus
lectin and ABC complex. The reaction products appear as dark brown deposits of DAB. The reaction
products for Lotus were localized in the cytoplasm and the brush borders of proximal tubules of
adult cortex. x 200. Bar=10μm
Edem, E. E.
15
HAVE A
NICE DAY!
Delve into the historical review of histochemistry, exploring its roots in the assessment of chemical compounds within cells and tissues using stains and dyes. Uncover the classification of histochemical methods, from chemical to biological approaches, and gain insights into the development and goals of this interdisciplinary field. Discover the techniques and methodologies employed in general and special histochemistry, encompassing diverse organ systems. By the end, grasp the essence of histochemistry as a marriage between biochemistry and histology.
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Presentation Transcript
Lecture 1: Historical Review on Histochemistry and Classification of Histochemical Methods Course Code: ANA 304 Course Title: General Histochemistry Course Lecturer: Edem E. Edem
Course Outline LECTURE ONE: 1.1 Historical review on histochemistry and cytochemistry 1.2 Classification of histochemical methods 1.2.1 Chemical methods 1.2.2 Physical methods 1.2.3 Biological methods LECTURE TWO: 2 Histological and Histochemical Techniques 2.1 Animal treatment and tissue processing 2.2 Fixation of tissues and cells 2.2.1 Chemical fixation for insoluble compounds 2.2.2 Chemical fixation for soluble compounds 2.2.3 Cryo-fixation for soluble compounds 2.3 Embedding and sectioning 2.3.1 Cryo-sectioning without embedding 2.3.2 Freeze-drying and embedding 2.3.3 Freeze-substitution and embedding 2.3.4 Dry sectioning of freeze-dried or freeze-substituted materials 2.4 Observation of histochemical specimens by microscopy 2.4.1 Light microscopy 2.4.2 Electron microscopy 2.4.3 Image analysis of histochemical reactions 2 Edem, E. E.
Course Outline LECTURE THREE: 3. Histochemical Methodology - General Histochemistry 3.1. Chemical methods in histochemistry 3.1.1. Color reactions for light microscopy 3.1.2. Electron dense deposits for electron microscopy 3.2. Physical methods in histochemistry 3.2.1. Methods using high or low temperatures 3.2.2. Methods using lights with different wave lengths 3.2.3. Methods using radiations LECTURE FOUR: 3.3. 3.3.1. 3.3.2. Biological methods in histochemistry Immunohistochemistry Lectin histochemistry 3 Edem, E. E.
Course Outline LECTURE FIVE: 4. Special Histochemistry - Histochemistry of the Organs 4.1. The skeletal system 4.2. The muscular system 4.3. The circulatory system 4.4. The digestive system LECTURE SIX: 4.5. 4.6. 4.7. 4.8. The respiratory system The urinary system The reproductive system The endocrine system LECTURE SEVEN: 4.9. 4.10. 5. Revision The nervous system The sensory system 4 Edem, E. E.
Objectives At the end of the class, students should be able to: Define histochemistry Elucidate the goals of histochemistry Appreciate the historical development of histochemistry as a scientific discipline Identify the related disciplines to histochemistry Classify the different methods used in histochemistry 5 Edem, E. E.
What is Histochemistry? Histo = histology: chemistry = chemical reactions The branch of science concerned with the qualitative and quantitative assessment of chemical compounds in a cell/tissue using stains/dyes and microscopy. It is a marriage between bio/chemistry and cyto/histology 6 Edem, E. E.
Historical Review on Histochemistry Histochemistry/cytochemistry is as old as histology itself In early 19th century, histochemical researches to study chemical components of biological structure in combination with chemistry and biology started first in botany in France (Raspail,1825). He stained starch in plant tissues blue with potassium iodide solution under the light microscope and demonstrated its localization microscopically Then, he published an essay on microscopic chemistry for the first time (Raspail, 1825) 7 Edem, E. E.
Historical Review on Histochemistry Later, from the 1840 s to 1870 s, histochemistry in zoology and medicine was developed mainly as biological chemistry, together with histology (Lehmann,1842). Among these zoologists, anatomists and pathologists who were at that time interested in analyzing chemical constituents in animal tissues including human, Miescher (1874) was the first to introduce cell fractionation to analyze nucleic acids in nuclei of leukocytes. During these times in the 19th century, this new field was called as microchemie in French or Mikrochemie in German, which meant microchemistry in English. 8 Edem, E. E.
Historical Review on Histochemistry Microchemistry was, in other words, microscopic chemistry or chemical microscopy and meant to observe chemical reactions in situ under microscopy. In early 20th century, aniline dyes were frequently used to stain tissues in anatomy and pathology. Histologists and pathologists were much interested in new dyes and less interested in histochemistry at that time. 9 Edem, E. E.
Historical Review on Histochemistry When Lison (1936) published his famous book entitled Histochemie Animale , many histologists were again interested in histochemistry. Lison classified histochemical techniques into two categories, m thodes extra situm or m thodes extractives and m thodes histochimiques in situ or m thodes topochimique in French. He proclaimed Histochemistry to be the new science 10 Edem, E. E.
Historical Review on Histochemistry Then, many histologists, anatomists, pathologists, physiologists started to study this new field and published many original papers and books dealing with histochemistry and cytochemistry. Innumerable literature, as well as textbooks and handbooks, are available from the 1950 s to 1990 s. Nagata (1995) proposed that the field should be designated General Histo-cytochemistry just like we have General Histology. One of the foremost histochemists in Nigeria is the Late Prof. Caxton-Martins 11 Edem, E. E.
Classification of Histochemical Methods Nagata (1995) proposed the classification of histochemical methods into: 1. Chemical methods such as chemical reactions by staining Light microscopic photograph of the colon of an adult mouse stained with HID-AB sequence. The goblet cells in the top region (upper half of the picture) of the crypts stained black with HID (high iron diamine), while the goblet cells in the deeper region (bottom) stained blue with AB (Alcian blue). x480. 12 Edem, E. E.
Classification of Histochemical Methods ii. Physical methods such as radiations Light microscopic radioautogram of the colon of a mouse injected with 35SO4, fixed at 180 min., sectioned, radioautographed and stained with toluidine blue. 13 Edem, E. E.
Classification of Histochemical Methods iii. Biological methods such as immunity Light micrograph of an adult mouse kidney cortex, cryosectioned, stained with biotinylated Lotus lectin and ABC complex. The reaction products appear as dark brown deposits of DAB. The reaction products for Lotus were localized in the cytoplasm and the brush borders of proximal tubules of adult cortex. x 200. Bar=10 m 14 Edem, E. E.
15 Edem, E. E.
HAVE A NICE DAY!
