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Human-Histology-Lecture-3

Histology is the study of body tissues and their organization into organs, focusing on the relationship between cell structure and function. Various microscopy techniques, including fluorescence, light, and electron microscopy, are employed to prepare and examine tissue samples, each with distinct methods and applications. Additionally, hybridization techniques are used to localize specific RNA or DNA sequences within tissues.

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10 views

Human-Histology-Lecture-3

Histology is the study of body tissues and their organization into organs, focusing on the relationship between cell structure and function. Various microscopy techniques, including fluorescence, light, and electron microscopy, are employed to prepare and examine tissue samples, each with distinct methods and applications. Additionally, hybridization techniques are used to localize specific RNA or DNA sequences within tissues.

Uploaded by

aleriasamson
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© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Human Histology (Lecture)

Histology and its methods

HISTOLOGY
● study of the tissues of the body and how
these tissues are arranged to constitute
organs.
● involves all aspects of tissue biology, with the
focus on how cells’ structure and arrangement
optimize functions specific to each organ.
● two interacting components: cells and
extracellular matrix (ECM).

PREPARATION OF TISSUES FOR STUDY

Fluorescence Microscopy

● Tissue sections are usually irradiated with


ultraviolet (UV) light and the emission is in the
visible portion of the spectrum.
● The fluorescent substances appear brilliant on
a dark background.
★ Fluorescence
→ Phenomenon wherein certain cellular
substances are irradiated by light of a proper
wavelength, they emit light with a longer wavelength.
● Acidic structures basic dye = basophilic (e.g.
nucleus)
● Basic structures acidic dye = acidophilic (e.g.
cytoplasm)
● Hematoxylin and eosin

MICROSCOPY

LIGHT MICROSCOPY

Bright-Field Microscopy
★ stained preparations are examined by means
of ordinary light that passes through the
specimen.
Phase-Contrast Microscopy
★ Resolving power
● based on the principle that light changes its
→ defined as the smallest distance between
speed when passing through cellular and
two particles at which they can be seen as separate
extracellular structures with different
objects.
refractive indices.
→ 0.2 μm
● uses a lens system that produces visible
images from transparent objects

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● enables examination of unstained cells and Confocal Microscopy
tissues and is especially useful for living cells. ● combines components of a light optical
microscope with a scanning system to dissect
Phase-Contrast Microscopy a specimen optically.
● Interference microscope ● Uses (1) a small point of high-intensity
→allows quantification of tissue mass light, often from a laser, and (2) a plate
with a pinhole aperture in front of the
● Differential interference microscope image detector.
→using Nomarski optics ● point light source, the focal point of the
→useful for assessing surface properties of lens, and the detector’s pinpoint aperture
are all optically conjugated or aligned to
cells and other biologic objects.
each other in the focal plane (confocal),
and unfocused light does not pass through
the pinhole.

Dark-field Microscopy
● no direct light from the light source is
gathered by the objective lens. Polarizing Microscopy
● only light that has been scattered or diffracted
● uses the fact that highly ordered molecules
by structures in the specimen reaches the
or arrays of molecules can rotate the angle
objective. of the plane of polarized light.
● equipped with a special condenser that ● Birefringence
illuminates the specimen with strong, oblique →ability to rotate the direction of
light. vibration of polarized light
● the field of view appears as a dark background
on which small particles in the specimen that
reflect some light into the objective appear
bright.

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ELECTRON MICROSCOPY

● based on the interaction of tissue components


with beams of electrons.
● The wavelength in the electron beam is much
shorter than that of light, allowing a 1000-fold
increase in resolution.
● 2 KINDS
○ TEM
○ SEM

Transmission Electron Microscopy Other Methods


● an imaging system that permits resolution
Enzyme Histochemistry
around 3 nm.
● magnifications of up to 400,000 times to be
viewed in detail ● A method for localizing cellular structures
● uses the interaction of a beam of electrons using a specific enzymatic activity present in
with a specimen to produce an image. those structures.
● Cryofracture and freeze etching
● a special method of sample preparation for
transmission electron microscopy
→it is especially important in the study of
membranes.

Scanning Electron Microscopy

Immunohistochemistry
● the electron beam does not pass through the
specimen but is scanned across its surface.
● the surface of the specimen is first dried and ● A highly specific interaction between
spray-coated with a very thin layer of heavy molecules is that between an antigen and its
metal (often gold) through which electrons do antibody.
not pass readily.
● When the beam is scanned from point to
point across the specimen, it interacts with
the metal atoms and produces reflected
electrons or secondary electrons emitted from
the metal

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HYBRIDIZATION TECHNIQUES

● a method of localizing messenger RNA


(mRNA) or DNA by hybridizing the
sequence of interest to a complementary
strand of a nucleotide probe.
● In situ hybridization
→binding of the nucleotide probe to the
DNA or RNA sequence of interest is performed
within cells or tissues, such as cultured cells or
whole embryos.

Interpretation of structures in tissue sections

4 Fionna Alexis B. Llamera

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