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94 Cards in this Set
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- Back
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Extra Cellular Matrix:
What is it? What makes it? Major Components: |
Macromolecular complex synthesized locally by fibroblasts that constitutes a large portion of any tissue.
Components: Proteoglycans & Glycosaminoglycans Adhesion Proteins: Fibronectin, Integrins, Laminin Collagen Elastic Fibers: Elastin, Fibrillin |
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Role of ECM
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1. Mechanical Support
2. Cell orientation 3. Control of cell growth 4. Cell differentiation 5. Establish tissue microenvironment 6. Storage and presentation to the cell of regulatory factors and growth factors |
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Carcinoma vs. Sarcoma
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Carcinoma: Cancer of the epithelium that has to break through the basal lamina to become invasive
Sarcoma: Malignancy of the Mesenchyme - already invasive |
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Macrophage
Derivation: Function: |
Derivation: Bone Marrow --> Monocyte --> Macrophage
Function: Phagocytosis of antibody-antigen complex Antigen presentation to lymphocytes Turnover of connective tissue ground substance Phagocytosis of apoptotic bodies |
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Found in loose connective tissue:
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1. Collagen
2. Fibroblasts 3. Lymphocytes - round with minimal to no cytoplasm 4. Plasma Cell 5. Macrophage - dark, granular cytoplasm around the nucleus 6. Adipose Cells |
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Found In Dense Regular Connective Tissue:
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1. Collagen
2. Fibroblasts |
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Method For Preparing Routine Samples For Light Microscopy
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1. Submission
2. Accessioning 3. Fixation - formalin 4. Dehydration - ethanol 5. Clearing - xylene 6. Embedding - wax 7. Sectioning 8. Mounting 9. Re-hydrating 10. Staining |
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What does a tissue sample represent?
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A 2D representation of a 3D structure
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Method for preparing EM samples
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1. Submission
2. Accessioning 3. Fixation 4. Dehydration 5. Clearing 6. Embedding 7. Sectioning 8. Mounting 9. Staining |
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4 Fundamental Tissues
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1. Epithelium
2. Nerve 3. Muscle 4. Connective Tissue |
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Role of formalin step
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1. Arrest cells in mitosis
2. Terminates cell metabolism 3. Prevents autolysis and decomposition 4. Kills pathogenic microorganisms |
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Method For Preparing Routine Samples For Light Microscopy
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1. Submission
2. Accessioning 3. Fixation - formalin 4. Dehydration - ethanol 5. Clearing - xylene 6. Embedding - wax 7. Sectioning 8. Mounting 9. Re-hydrating 10. Staining |
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What is preserved in the tissue sample after clearing?
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Large molecules:
1. Nucleoproteins, DNA, mRNA 2. Intermediate filaments, microtubules, collagen 3. membrane phospholipid, glycolipid |
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What does a tissue sample represent?
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A 2D representation of a 3D structure
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What is washed out of the tissue sample after clearing?
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Small molecules and soluble components
1. tRNA 2. Glycogen 3. Neutral Lipids |
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Method for preparing EM samples
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1. Submission
2. Accessioning 3. Fixation 4. Dehydration 5. Clearing 6. Embedding 7. Sectioning 8. Mounting 9. Staining |
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Method for preparing frozen section samples:
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1. Submission
2. Accessioning 3. Embedding - Cryoprotectant 4. Sectioning - Cryostat 5. Mounting - glass slides 6. Staining - H&E, Oil Red O |
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4 Fundamental Tissues
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1. Epithelium
2. Nerve 3. Muscle 4. Connective Tissue |
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Stain for Carbohydrates
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1. PAS
2. Mucicarmine |
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Role of formalin step
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1. Arrest cells in mitosis
2. Terminates cell metabolism 3. Prevents autolysis and decomposition 4. Kills pathogenic microorganisms |
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Stain for connective tissue
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1. Trichrome
2. Reticulin 3. Elastin |
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What is preserved in the tissue sample after clearing?
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Large molecules:
1. Nucleoproteins, DNA, mRNA 2. Intermediate filaments, microtubules, collagen 3. membrane phospholipid, glycolipid |
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What is washed out of the tissue sample after clearing?
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Small molecules and soluble components
1. tRNA 2. Glycogen 3. Neutral Lipids |
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Method for preparing frozen section samples:
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1. Submission
2. Accessioning 3. Embedding - Cryoprotectant 4. Sectioning - Cryostat 5. Mounting - glass slides 6. Staining - H&E, Oil Red O |
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Stain for Carbohydrates
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1. PAS
2. Mucicarmine |
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Stain for connective tissue
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1. Trichrome
2. Reticulin 3. Elastin |
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Stain for Amyloid
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1. Congo Red
2. Sirius Red |
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Stain for fat
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1. Sudan B
2. Oil Red O |
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Stain for microorganisms
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1. AFB - mycobacteria
2. GMS - fungus 3. Gram - bacteria |
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Stain for nervous system
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1. Luxol Blue
2. Cajal 3. Nissl |
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Stain for pigments and minerals
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1. Iron - prussian blue
2. Calcium - von kossa 3. Copper - rhodabine |
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Cytoplasmic Inclusions
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1. Hemosiderin - iron
2. Lipofuscin - Wear and tear 3. Glycogen - Around the mitochondria |
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Filament type in Epithelia
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Cytokeratins
Examples? |
Keratinizing and non-keratinizing epithelia
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Filament type in muscle
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Desmin
Examples? |
Smooth and striated muscle
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Filament Type in Mesenchymal Fibroblasts
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Vimentin
Examples? |
Chondroblasts, macrophages, endothelial cells, vascular smooth muscle
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Filament Type in Glial Cells
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Glial Fibrillary Acidotic Proteins
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Astrocytes
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Filament type in neurons
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Neurofilaments
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Nerve cell body and processes
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Examples of Microfilaments
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Actin and Myosin
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Examples of intermediate filaments
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Cytoskeleton (also made of microtubules, thin filaments, centrioles)
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Microvilli - function and composition
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Function - Increase surface area for absorption
Composition - Actin (microfilaments) |
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Examples of Microtubules
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Components of cilia
Made of tubulin |
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Cilia - function and composition
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Function - Moving substances out of the respiratory tract
Composition - Tubulin (microtubules) |
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Golgi Apparatus Function
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Proteins enter at the Cis face from the RER.
Proteins exit through the Trans face. They exit inside secretory vesicles. |
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Smooth ER function
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1. Detoxification
2. Production of cholesterols, steroids, and lipids 3. Sequesters Ca++ in muscles |
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Rough ER function
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Synthesis and modification of proteins for packaging
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Difference between LM and EM preparation
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1. EM does not involve a rehydration step because it requires a very thin sample that electrons may pass though.
2. Mounting - copper matrix 3. Fixation - Glutaraldehyde and OsO4 4. Clearing - Propylene Oxide 5. Embedding is done with plastic 6. Sectioning – ultramicrotome 7. Staining – lead citrate, uranyl acetate |
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FISH (Fluorescence In Situ Hybridization
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Probes, which are segments of clones DNA labeled with a fluorescent tag and hybridized to chromosomal DNA. This can be done during interphase, which is where cells spend most of their time.
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Pars Amorpha
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Where rRNA is transcribed
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Pars Fibrosa
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Filled with rRNA transcripts
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Pars Granulosa
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Mature ribosomal subunits
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Cyclic Alterations between mitosis and interphase.
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G0 – cell is out of the cell cycle, but can be bumped back in
G1 – cells get bigger – check point (yes, duplicate) S Phase – DNA replication G2 – Cells get bigger again – check point (yes, duplicate) M - Mitosis |
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Geimsa Stain
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Blood and Bone Marrow Stain
Can differentiate white and red blood cells |
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Direct vs. Indirect Immunocytochemistry
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Direct – An antibody is deployed against a certain molecule
Indirect – An antibody is deployed against an antibody of a certain molecule |
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Pinocytosis
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Cellular drinking and small molecules
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Phagocytosis
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Ingestion of large particles such as cell debris, bacteria, and other foreign materials
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Receptor-mediated endocytosis
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Allows entry of specific molecules into the cell
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Describe the structural organization of Centrioles and their relationship to microtubules.
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Microtubules form around centrioles and position the mitotic spindle properly
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Apoptosis vs. Necrosis
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Apoptosis - Programmed cell death. Highly regulated process in which cells rapidly produce small membrane-enclosed apoptotic bodies, which undergo phagocytosis by neighboring cells or macrophages
Necrosis – Result of accidental injury. The cell swells and their organelles increase in volume. They burst, realeasing their contents into the extracellular space. Immunodefensive cells are prompted by macrophages to promote inflammation. |
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Describe how dysfunction of cell cycle checkpoints can lead to tumor cell production.
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At G1, the DNA can be fixed before starting S phase, but if the gene encoding p53 is damaged, then recognition of DNA damage is dysfunctional, and damaged DNA is replicated
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How are actin filaments in microvilli cross-linked to each other?
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Fimbrin and Villin
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What is the principal function of the zonula adherens?
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Zonula Adherens function is to give rigidity and cohesion to an epithelium
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The basal lamina can only be seen by EM. What stain does EM use?
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Uronyl Acetate and Lead Acetate
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What are desmosomes made of and what is their function?
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Desmocolin and desmoglein
Function: Resist shearing forces |
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2 Kinds of connective tissue proper
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Loose and Dense Connective Tissue
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Which cells are resident cells of connective tissue?
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1. Fibroblasts
2. Neutrophils 3. Macrophages 4. Lymphocytes (not permanent residents) |
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What differentiates into a macrophage?
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Monocyte
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What is a macrophage called in
skin? |
Langerhans
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What is a macrophage called in liver?
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Kuppfer
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What is a macrophage called in Lungs?
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Dust Cells
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What is a macrophage called in Brain?
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Microglia
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What is a macrophage called in connective tissue?
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1. Multinuclear Giant Cell
2. Macrophage |
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What is a macrophage called in Blood?
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Monocyte
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What is a macrophage called in bone?
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Osteoclast
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What is a macrophage called in Lymph nodes?
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Dendritic Cell
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What is a Barr Body?
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Genetically inactive X chromatin that can be seen in the buccal epithelium.
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What are epithelioid cells?
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Groups of Macrophages or Multinuclear Giant Macrophages that are aggregated when there is pathology. They look like epithelium.
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What differentiates into a mast cell?
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Basophil
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Mast Cell Function and Structure
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Function: They release granules upon exposure to antigens. Involved in histamine reaction.
Found - near blood vessels Function - Participate in allergic reactions |
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What differentiates into a plasma cell?
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Lymphocyte
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Eosinophils - Structure and Function
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Structure - Large granules that stain red with H&E and a characteristic BI-LOBED nucleus.
Function - They phagocytose antigen-antibody complexes and mediate allergic reactions and asthma. |
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Neutrophils - Structure and Function
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Structure - Granules that stain lightly and 2-5 nuclear lobes linked by thin extensions. Contain lysosomes and barr bodies.
Function - Break down engulfed bacteria with lysosomes. They clean up enflamed and necrotic tissue. They are short lived. |
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Where is mucous connective tissue usually found?
What does it resemble? |
In the umbilical cord.
It resembles embryonic mesenchyme. |
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Stains that let you study adipose tissue.
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1. Oil Red O - used during cryostat
2. Osmium Tetroxide - preserves the lipid 3. H&E - shows the vacuole that is left after clearing. |
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Where are white adipose cells found?
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In loose or irregular dense connective tissue.
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What hormone does unilocular adepocyte release?
What is the function of this hormone? |
Hormone = Leptin
Function of Leptin = It is the satiety response. It regulates the amoung of adipose tissue and reulates the appetite under normal condition. |
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Where would you find white adipose tissue in a person who was undernourished?
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Adipose tissue in the palms, soles, and retroorbital fat pads.
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Stain for connective tissue
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Mallory Trichrome
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Pararosaniline - Toluidine Blue Stain
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Chromatin stains purple and cytoplasm and collagen stain violet
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Where is brown fat located?
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1. Around the kidney
2. Mediastinum 3. Adrenal Glands 4. Aorta |
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Stain for blood
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Giemsa
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Stain for Collagen Type III
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EVG - silver stain
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Stain for Lipids
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Sudan Black
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What organelles can only be seen with EM?
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Ribosomes
Smooth ER Plasma Membrane Endosomes |
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Resident Cells
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1. Fibroblasts
2. Adipocytes 3. Macrophages 4. Mast Cells 5. Adult Stem Cells |
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