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(Dehydrating Solutions), Ending 100%, (Small Intestine Under H&E Stain)

The document provides an overview of the process of preparing tissue samples for histological study under a microscope. Key steps include: 1) Fixation of tissues using chemicals to preserve cell structure, 2) Dehydration by transferring tissues through increasingly concentrated alcohol solutions, 3) Clearing tissues using organic solvents, 4) Infiltration of tissues with paraffin wax, 5) Embedding tissues in paraffin blocks, 6) Sectioning tissues using a microtome, 7) Staining sections with dyes such as hematoxylin and eosin for visualization, and 8) Mounting stained sections on slides for examination under light or electron microscopes.

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yuli
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51 views2 pages

(Dehydrating Solutions), Ending 100%, (Small Intestine Under H&E Stain)

The document provides an overview of the process of preparing tissue samples for histological study under a microscope. Key steps include: 1) Fixation of tissues using chemicals to preserve cell structure, 2) Dehydration by transferring tissues through increasingly concentrated alcohol solutions, 3) Clearing tissues using organic solvents, 4) Infiltration of tissues with paraffin wax, 5) Embedding tissues in paraffin blocks, 6) Sectioning tissues using a microtome, 7) Staining sections with dyes such as hematoxylin and eosin for visualization, and 8) Mounting stained sections on slides for examination under light or electron microscopes.

Uploaded by

yuli
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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INTRODUCTION TO HISTOLOGY 7.

Trimming
- paraffin block is trimmed to expose the
Histology tissue for sectioning (slicing) on a
- the study of the tissues of the body and microtome
how these tissues are arranged to
constitute organs 8. Tissues Sectioning
- sectioning (slicing) of tissue using a
PREPARATION OF TISSUES FOR microtome
STUDY - placed in floatation water bath to
minimally melt the paraffin
Biopsy: tissue examination from alive
Autopsy: tissue examination from dead 9. Staining
- addition of staining dyes to visualize the
1. Gross Examination tissue structures under the microscope
- tissue samples from the organ - most common:
- describing the specimen Hematoxylin and Eosin dye (H&E)
- placing all or parts of it into a small - acidophilic and basophilic dyes
plastic tissue cassette which holds the
tissue while it is being processed to a 10. Mounting
paraffin block - addition of mounting medium to the
stained tissues and cover with cover slip
2. Fixation to preserve the tissues
- small pieces of tissue are placed in - adhesive and preservative
solutions of chemicals (fixatives)
- cross-link proteins and inactivate
degradative enzymes, which preserves
cells and tissue structure
- most common fixative: formaldehyde

3. Dehydration
- tissue is transferred through a series of
increasingly concentrated alcohol solutions
(dehydrating solutions), ending 100%,
which removes all water
- increasing concentration dapat (from (small intestine under H&E stain)
30%, 50%, 70%, 90%, absolute alcohol)
- most common dehydrator: ethanol MICROSCOPY
- remove water since we are dealing with
water insoluble solutions Two major categories:

4. Clearing A. LIGHT MICROSCOPY


- alcohol is removed in organic solvents
(clearing agents) in which both alcohol  Bright-Field Microscopy
and paraffin are miscible - stained tissue is examined with
- remove alcohol ordinary light passing through the
- commonly used: xylene (5 series) preparation

5. Infiltration
- tissue is the placed in melted paraffin
(impregnating agent) until it becomes
completely infiltrated with this substance
- using paraffin oven

6. Embedding
- paraffin-infiltrated tissue is placed in a
small mold with melted paraffin and
allowed to harden
- paper boat mold sa pinas bc pobre
 Fluorescence Microscopy  Polarizing Microscopy
- tissue sections are usually irradiated - produces an image only of material
with ultraviolet (UV) light and the having repetitive, periodic
emission is in the visible portion of the macromolecular structure
spectrum - strike a specific pathway
- fluorescent substances (dyes or
lamps) appear bright on a dark
background
- has a source of UV or other light and
filters that select rays of different
wavelengths****

B. ELECTRON MICROSCOPY
- supreme

 Transmission Electron
Microscopy (TEM)
- magnified as much as 400,000 times
to be viewed in detail

 Phase-Contrast Microscopy
- uses a lens system that produces
visible images from transparent
objects
- unstained preparation
- based on the principle that light
changes its speed when passing
through cellular and extracellular
structures*****

 Scanning Electron Microscopy


- produces and focuses a very narrow
 Confocal Microscopy
beam of electrons, but in this
- high resolution and sharp focus than
instrument the beam does not pass
bright-field microscopy
through the specimen
- computer aided microscope
- the surface of the specimen is first
- eliminate stray and scattered light
dried and spray-coated with a very
using confocal pinholes and splitter
thin layer of heavy metal (often gold)
which reflects electrons in a beam
scanning the specimen
- reflected electrons are captured by a
detector, producing signals that are
processed to produce b&w images
- computer aided

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