Activity 1-6 - From histology PDF

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Warning: TT: undefined function: 32MICROSCOPE Optical instrument which magnifies the size of the object Used to observed specimens that are too small to be seen by the naked eye Invention was credited to Galileo Galilei and Zacharias Janssen Its use started with Anton Van Leuwenhoek Simple Microscop...

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HISTOLOGY LABO LABORATORY RATORY

Ms. Rogie Mae G. Apelarta & Ms. Lory Ann Braza

ACTIVITY 1: THE BRIGHTFIELD MICROSCOPE MICROSCOPE

LIGHT MICROSCOPE 1.

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Optical instrument which magnifies the size of the object

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Used to observed specimens that are too small to be seen by the naked eye

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Invention was credited to Galileo Galilei and Zacharias Janssen

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Its use started with Anton Van Leuwenhoe k

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Types of Microscope according to number of Lens

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Types of Compound Microscope according to Illumination 1. LIGHT MICROSCOPE – employs visible light as source of illumination

2.

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Brightfield Darkfield

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Fluorescence Phase-Contrast Confocal

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Polarizing

ELECTRON MICROSCOPE – employs beam of electrons as a source

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Objects appear dark against a bright background Can be converted to Darkfield microscope by changing its CONDENSER

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Can be converted to polarizing microscope by adding 2 polarizing filters

DARKFIELD MICROSCOPE ‫ ﻼ‬Objects appear bright against a dark background ‫ﻼ‬ ‫ﻼ‬

1. Simple Microscope  Has a single biconvex magnifying lens 2. Compound Microscope  Has 2 or more lenses  Classified based on illumination

BRIGHTFIELD MICROSCOPE ‫ ﻼ‬Method most commonly/routinely used

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Routinely used to observe Treponema pallidum For spirochetes  Treponema  Borrelia  Leptospira

FLUORESCENECE MICROSCOPE ‫ ﻼ‬uses ultraviolet light that when strikes the organism can cause them to glow against a dark background. ‫ﻼ‬ ‫ﻼ‬

usually used to demonstrate Ag-Ab reactions commonly used to know the presence of autoantibodies (e.g. Anti-Nuclear Antibody)

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Cyclospora cayetanensis – naturally fluorescing parasite

CONFOCAL MICROSCOPE ‫ﻼ‬

Involves scanning the specimen at successive focal planes with a focused light beam, often from a laser, and produces a 3D reconstruction from the images.

of illumination • Transmission EM (TEM) • Scanning EM (SEM)

MLS 2B AY 2019-2020

MUYCO, Ma. Christy M.

HISTOLOGY LABO LABORATORY RATORY

5.

POLARIZING MICROSCOPE ‫ ﻼ‬Uses 2 polarizing filters ‫ ﻼ‬designed to observe and photograph specimens that are

Ms. Rogie Mae G. Apelarta & Ms. Lory Ann Braza

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STAGE – platform with 2 clips which holds the slide to be studied

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MECHANICAL STAGE - attached to the stage for moving the slides

visible primarily due to their optically anisotropic character ELECTRON MICROSCOPE 1.

2.

TRANSMISSION EM (TEM) ‫ﻼ‬

Produces 2-dimensional images of the cell’s internal structure.

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Taller column compared to SEM Electron beam pass through the sample

OPTICAL PARTS 

MIRROR - used to gather & direct light for illumination

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LIGHT SOURCE – houses the light needed for illumination

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IRIS DIAPHRAGM – controls the amount of light reaching the object

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CONDENSER – concentrates the light rays into the specimen

SCANNING EM (SEM) ‫ ﻼ‬Produces 3-dimensional images of the cell’s outer structure. ‫ ﻼ‬Short column than TEM ‫ﻼ‬

Electron beam scans though the surface

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stage. 

OCULAR – aka eyepiece, found in the draw tube, used for further

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OBJECTIVES:

magnification of the object.

PARTS OF THE MICROSCOPE MECHANICAL PARTS

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BASE - “Y” – shaped on which the microscope stands

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PILLAR – short supporting piece arising from the base

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ARM – curved handle used for carrying the microscope

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INCLINATION JOINT – joins the pillar & arm area

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BODY TUBE – attached to the arm & which bears the lenses

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DRAW TUBE – bears the eyepiece

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REVOLVING NOSEPIECE – holds the objectives

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DUST SHIELD – protects dust from shielding the objectives, found

The iris diaphragm & the condenser constitutes the sub

SCANNER – shortest lens screwed to the revolving nosepiece which serves for primary focusing

on the upper part of the revolving nosepiece 

COARSE ADJUSTMENT KNOB – big knob, brings object to focus

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FINE ADJUSTMENT KNOB – small knob, for more delicate focusing MLS 2B AY 2019-2020

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LOW- POWER OBJECTIVE (LPO) – shorter lens, which serves to bring the object into focus.

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HIGH-POWER OBJECTIVE (HPO) – longer lens which serves to form a bigger image of the object for further studies

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OIL-POWER OBJECTIVE (OIO) – longest lens which serves to bring a more detailed aspect of the object being studied, usually needs special oil

MUYCO, Ma. Christy M.

HISTOLOGY LABO LABORATORY RATORY

Ms. Rogie Mae G. Apelarta & Ms. Lory Ann Braza

TOTAL MAGNIFICATION

OBJECTIVE LENSES Scanning lens (Lowest; Red mark) LPO (Yellow mark) HPO (Blue mark) OIO (White mark)

MAGNIFICATION of OCULAR LENS

MAGNIFICATION TOTAL of OBJECTIVE MAGNIFICATION LENS

10

4

40x

10

10

100x

10

40

400x

10

100

1000x

HANDLING AND CARE of MICROSCOPE 1.

Carry the microscope with two hands, one beneath the base and the other hand grasping the arm, in upright position.

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Place the microscope carefully on the working table, about 1 inch away from the edge of the table

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Remove the dust from the microscope using soft brush (preferably Camel’s hair) or you may blow it away. Only after this should the lenses are cleaned with lens paper

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Clean the eyepiece lens with dry lens paper and the objective lenses with lens paper or cotton (Dry or moistened with xylene or 95% ethyl alcohol).

MAGNIFICATION and RESOLUTION MAGNIFICATION •

refers to the amount or degree of visual enlargement of an observed object. Magnification is measured by multiples, such as 2x, 4x and 10x, indicating that the object is enlarged to twice as big, four times as big or 10 times as big, respectively.

RESOLUTION •

defined as the shortest distance between two points on a specimen that can still be distinguished by the observer or camera system as separate entities MLS 2B AY 2019-2020

MUYCO, Ma. Christy M.

HISTOLOGY LABO LABORATORY RATORY

Ms. Rogie Mae G. Apelarta & Ms. Lory Ann Braza

ACTIVITY 2: THE ANIMAL CELL 5.

If the microscope is equipped with electric cord, check if it is in good condition before plugging into the outlet

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Make sure that the scanner or the low power objective (LPO) is in focusing position. Always start each new observation at scanner or LPO. Start with lowest power objective.

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If the microscope is equipped with a mirror, use lamp or sunlight as source of light

CELL • •

• NOTE: -

If you wear a glasses, take them off

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If you see only your eyelashes, move closer

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If you see a dark line that goes part way across the field view, try turning the eyepiece o Or diaphragm is partially open

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Use only fine adjustment knob when using HPO o Because when you use course adjustment knob can cause scratch on lens or the slide will be broken.

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As much as possible keep both eyes open to reduce eyestrain.

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Return the objectives to the lowest power after use.

Basic structural and functional unit of life There are two types: o Prokaryotic Cell – does not have membrane-bounded nucleus and organelles o Eukaryotic Cell – has distinct nucleus and complex membranous organelles in cytoplasmic matrix and their genetic materials (DNA) are within the membrane-bounded nuclei Principal parts of a cell are the plasma (cell) membrane, cytosol, organelles, and inclusions.

HOW TO DISINFECT MICROSCOPE -

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In a clean piece of cloth and spray the Lysol and wipe it on the microscope, especially on the adjustment knob, arm, revolving nose piece Also the stage should be disinfected

MLS 2B AY 2019-2020

MUYCO, Ma. Christy M.

HISTOLOGY LABO LABORATORY RATORY

Ms. Rogie Mae G. Apelarta & Ms. Lory Ann Braza

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The long period between mitoses (the G1, S, and G2 phases) is also commonly called interphase.

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During the relatively long prophase, several changes occur: o The nucleolus disappears and the replicated chromatin condenses into discrete threadlike chromosomes, each consisting of duplicate sister chromatids joined at the centromere.

GENERALIZED HUMAN CELL

The two centrosomes with their now-duplicated centrioles separate and migrate to opposite poles of the cell and organize the microtubules of the mitotic spindle.

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Late in prophase, lamins and inner nuclear membrane are phosphorylated, causing the nuclear lamina and nuclear pore complexes to disassemble and disperse in cytoplasmic membrane vesicles.

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During metaphase , chromosomes condense further and large protein complexes called kinetochores, located at the centromere DNA region of each chromosome, attach to the mitotic spindle. The cell is now more spherical and microtubules move the chromosomes into alignment at the equatorial plate.

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In anaphase, sister chromatids (now called chromosomes themselves) separate and move toward opposite spindle poles by a combination of microtubule motor proteins and dynamic changes in the lengths of the microtubules as the spindle poles move farther apart.

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At telophase, the following occur: o The two sets of chromosomes are at the spindle poles and begin reverting to their uncondensed state.

CELL DIVISION During mitosis, a parent cell divides and each of the two daughter cells receives a chromosomal set identical to that of the parent cell. The chromosomes replicated during the preceding S phase are distributed to the daughter cells.

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Microtubules of the spindle depolymerize and the nuclear envelope begins to reassemble around each set of daughter chromosomes.

MUYCO, Ma. Christy M.

HISTOLOGY LABO LABORATORY RATORY

o

Ms. Rogie Mae G. Apelarta & Ms. Lory Ann Braza

A belt-like contractile ring of actin filaments associated with myosins develops in the cortical cytoplasm at the cell’s equator. During cytokinesis at the end of telophase, constriction of this ring produces a cleavage furrow and progresses until the cytoplasm and its organelles are divided into two daughter cells, each with one nucleus.

Each phase of the cell cycle has one or more checkpoints where the quality of specific cell activities is checked. Progression to the next phase of the cycle does not occur until all activities of the preceding phase are completed satisfactorily. Three important checkpoints are shown here, including

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The cell cycle is divided into interphase (blue) and cell division (mitosis and cytokinesis). Interphase is divided into G1, S, and G2 subphases. During G1, the cell carries out routine metabolic activities. During the S phase, DNA is replicated. During the G2 phase, the cell prepares for division. Following mitosis, two cells are formed by the process of cytokinesis. Each new cell begins a new cell cycle. Many cells exit the cell cycle and enter the G0 phase, where they remain until stimulated to divide, at which point they reenter the cell cycle.

MLS 2B AY 2019-2020

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The start or restriction checkpoint just before the start of S

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The G2/M checkpoint that ensures that DNA replication is complete

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The metaphase spindle checkpoint that ensures that all chromosomes will be segregated

Overall progression in the cycle is regulated by proteins called cyclins and cyclin-dependent kinases (CDKs) that phosphorylate/ activate enzymes and other proteins needed for phasespecific functions.

MUYCO, Ma. Christy M.

HISTOLOGY LABO LABORATORY RATORY

Ms. Rogie Mae G. Apelarta & Ms. Lory Ann Braza

ACTIVITY 3: THE EPITHELIAL TISSUES

EPITHELIAL TISSUE

TISSUES • • •

Aggregate of similar cells and cell products of common origin and function French word tisser means weave, is an apt word since tissues comprise fabric of organs Categorized into 4 main types: o Epthelial, Connective, Muscular & Nervous TISSUE

Epithelial

Connective

Muscle

CELLS Aggregated polyhedral cells

Several types of fixed and wandering cells Elongated contractile cells

Nervous

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EXTRACELLULAR MATRIX Small amount

Abundant amount Abundant amount

FUNCTIONS Lining of surface or body cavities; glandular secretion Support and protection of tissues/organs Strong contraction; body movements Transmission of nerve impulses

Elongated cells Very small with extremely amount fine processes Connective tissue is characterized by cells producing very abundant ECM; muscle tissue is composed of elongated cells specialized for contraction and movement; and nervous tissue is composed of cells with long, fine processes specialized to receive, generate, and transmit nerve impulses.

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Composed of compactly arranged cells with little intercellular component between them.

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Present in 2 forms : 1. Epithelium – a sheet of contiguous cells that covers the body surfaces and lines the body cavities 2.

Glands – which secretory, sensory & reproductive functions

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Has one side unattached or exposed called “free surface” and the other side is generally anchored to underlying connective tissue by a basement membrane

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Epithelia are classified according to morphology of the cells and number of cell layers.

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Morphology: squamous (flat), cuboidal, columnar or transitional

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Cell Layers: single layer (simple epithelium), many layers (stratified epithelium), some may appear to be stratified but is not (pseudostratified epithelium) SIMPLE EPITHELIUM

SIMPLE SQUAMOUS EPITHELIUM

Most organs can be divided into the parenchyma, which is composed of the cells responsible for the organ’s specialized functions, and the stroma, the cells of which have a supporting role in the organ. Except in the brain and spinal cord, the stroma is always connective tissue. MLS 2B AY 2019-2020

MUYCO, Ma. Christy M.

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Structure: Single layer of flat, often hexagonal cells; the nuclei appear as bumps when viewed as a cross section because the cells are so flat

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Function: Diffusion, filtration, some secretion, and some protection against friction

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Ms. Rogie Mae G. Apelarta & Ms. Lory Ann Braza

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Location: Kidney tubules, glands and their ducts, choroid plexuses of the brain, lining of terminal bronchioles of the lungs, surfaces of the ovaries

SIMPLE COLUMNAR EPITHELIUM

Location: Lining of blood vessels and the heart, lymphatic vessels (endothelium) and small ducts, alveoli of the lungs, portions of the kidney tubules, lining of serous membranes (mesothelium) of the body cavities (pleural, pericardial, peritoneal), and inner surface of the tympanic membranes

SIMPLE CUBOIDAL EPITHELIUM

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Structure: Single layer of cubeshaped cells; some cells have microvilli (kidney tubules) or cilia (terminal bronchioles of the lungs)

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Function: Secretion and absorption by cells of the kidney tubules; secretion by cells of glands and choroid plexuses; movement of particles embedded in mucus out of the terminal bronchioles by ciliated cells

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Structure: Single layer of tall, narrow cells; some cells have cilia (bronchioles of lungs, auditory tubes, uterine tubes, and uterus) or microvilli (intestines)

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Function: Movement of particles out of the bronchioles of the lungs by ciliated cells; partially responsible for the movement of oocytes through the uterine tubes by ciliated cells; secretion by cells of the glands, the stomach, and the intestines; absorption by cells of the small and large intestines

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Location: Glands and some ducts, bronchioles of the lungs, auditory tubes, uterus, uterine tubes, stomach, intestines, gallbladder, bile ducts, ventricles of the brain

MUYCO, Ma. Christy M.

HISTOLOGY LABO LABORATORY RATORY

Ms. Rogie Mae G. Apelarta & Ms. Lory Ann Braza

 NON-KERATINIZED STRATIFIED EPITHELIUM STRATIFIED SQUAMOUS EPITHELIUM  KERATINIZED

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Structure: Multiple layers of cells that are cube-shaped in the basal layer and progressively flattened toward the surface. In keratinized stratified epithelium, the cytoplasm of cells at the surface is replaced by a protein called keratin, and the cells are dead o Differentiating cells become keratinized, that is, filled with keratin and other substances, eventually lose their nuclei and organelles, and form superficial layers flattened squames that impede water loss. o Keratinized cells are sloughed off and replaced by new cells from more basal layers, which are discussed fully with the skin Function: helps protect against water loss across this epithelium.

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Structure: Multiple layers of cells that are cube-shaped in the basal layer and progressively flattened toward the surface, in nonkeratinized stratified squamous epithelium, the surface cells retain a nucleus and cytoplasm.

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Function: Protection against abrasion, a barrier against infection, reduction of water loss from the body

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Location: nonkeratinized—mouth, throat, larynx, esophagus, anus, vagina, inferior urethra, cornea

STRATIFIED CUBOIDAL EPITHELIUM

Location: found mainly in the epidermis of skin, where it helps prevent dehydration from the tissue

MLS 2B AY 2019-2020

MUYCO, Ma. Christy M.

HISTOLOGY LABO LABORATORY RATORY

Ms. Rogie Mae G. Apelarta & Ms. Lory Ann Braza

PSEUDOSTRATIFIED COLUMNAR EPITHELIUM AND TRANSITIONAL •

Structure: Multiple layers of somewhat cube-shaped cells

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Function: Secretion, absorption, protection against infection

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Location: Sweat gland ducts, ovarian follicular cells, salivary gland ducts

PSEUDOSTRATIFIED COLUMNAR EPITHELIUM

STRATIFIED COLUMNAR EPITHE...