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Questions and Answers
Which of the following is NOT a common histological stain?
Which of the following is NOT a common histological stain?
What is the primary function of Hematoxylin in H&E staining?
What is the primary function of Hematoxylin in H&E staining?
If a tissue sample appears bright pink after H&E staining, which of the following cell structures is likely enriched in the tissue?
If a tissue sample appears bright pink after H&E staining, which of the following cell structures is likely enriched in the tissue?
Which of the following stains would be most appropriate for examining the distribution of elastic fibers in cartilage?
Which of the following stains would be most appropriate for examining the distribution of elastic fibers in cartilage?
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Which of the following microscopy techniques would be most suitable for examining a slide stained with a fluorescent antibody (immunostaining)?
Which of the following microscopy techniques would be most suitable for examining a slide stained with a fluorescent antibody (immunostaining)?
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Which stain would be most suitable for examining glycogen concentration in the liver?
Which stain would be most suitable for examining glycogen concentration in the liver?
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Which of the following cell structures would be most prominently visible in a cell that is actively synthesizing proteins?
Which of the following cell structures would be most prominently visible in a cell that is actively synthesizing proteins?
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Which of the following staining methods is most commonly used for routine histological examination?
Which of the following staining methods is most commonly used for routine histological examination?
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Which of the following cellular structures would be most clearly visualized using a dark-field microscope?
Which of the following cellular structures would be most clearly visualized using a dark-field microscope?
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What is the main purpose of H&E staining in histology?
What is the main purpose of H&E staining in histology?
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Which of the following cell types are likely to have abundant mitochondria?
Which of the following cell types are likely to have abundant mitochondria?
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What is the main reason why smooth ER does not stain intensely in H&E staining?
What is the main reason why smooth ER does not stain intensely in H&E staining?
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What can you infer about a cell with a large, pale-staining region near the nucleus in H&E staining?
What can you infer about a cell with a large, pale-staining region near the nucleus in H&E staining?
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Why does rough ER stain basophilic and purple in H&E staining?
Why does rough ER stain basophilic and purple in H&E staining?
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What is the primary function of mitochondria?
What is the primary function of mitochondria?
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In H&E staining, why do cells with an abundance of mitochondria have a pink cytoplasm?
In H&E staining, why do cells with an abundance of mitochondria have a pink cytoplasm?
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Which of the following statements is TRUE about secretory vesicles?
Which of the following statements is TRUE about secretory vesicles?
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Which of the following statements is TRUE about the Golgi apparatus?
Which of the following statements is TRUE about the Golgi apparatus?
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Which of the following structures would be most prominent in cells that are actively synthesizing proteins?
Which of the following structures would be most prominent in cells that are actively synthesizing proteins?
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Which of the following is NOT a component of the cytoskeleton?
Which of the following is NOT a component of the cytoskeleton?
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Which of the following cytoskeletal components is primarily involved in cell movement?
Which of the following cytoskeletal components is primarily involved in cell movement?
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Which of the following is a common protein found in intermediate filaments?
Which of the following is a common protein found in intermediate filaments?
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Which of the following microscopy techniques would be required to visualize the cytoskeleton?
Which of the following microscopy techniques would be required to visualize the cytoskeleton?
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Which type of cell inclusion is stained with Periodic Acid-Schiff (PAS) reaction?
Which type of cell inclusion is stained with Periodic Acid-Schiff (PAS) reaction?
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Cells with a high level of RER and secretory granules are likely specialized for which function?
Cells with a high level of RER and secretory granules are likely specialized for which function?
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Which of the following is TRUE about cells with very basophilic cytoplasm and euchromatic nuclei?
Which of the following is TRUE about cells with very basophilic cytoplasm and euchromatic nuclei?
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Which of the following statements accurately describes the appearance of the plasma membrane in a hematoxylin and eosin (H&E) stained section?
Which of the following statements accurately describes the appearance of the plasma membrane in a hematoxylin and eosin (H&E) stained section?
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What makes the nucleus appear purple in H&E staining?
What makes the nucleus appear purple in H&E staining?
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Which of the following features is associated with a cell that exhibits an abundant supply of ribosomes in its cytoplasm?
Which of the following features is associated with a cell that exhibits an abundant supply of ribosomes in its cytoplasm?
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What is the primary advantage of using Transmission Electron Microscopy (TEM) to visualize cellular structures over Hematoxylin and Eosin (H&E) staining?
What is the primary advantage of using Transmission Electron Microscopy (TEM) to visualize cellular structures over Hematoxylin and Eosin (H&E) staining?
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Which of the following statements best describes the function of the endoplasmic reticulum (ER)?
Which of the following statements best describes the function of the endoplasmic reticulum (ER)?
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Flashcards
Methods of staining
Methods of staining
Techniques used to enhance contrast in biological tissues for microscopic examination.
H&E staining
H&E staining
Hematoxylin and eosin staining, a widely used technique in histology for tissue analysis.
Microscope for H&E
Microscope for H&E
A light microscope is typically used to examine slides stained with H&E.
Fluorescent antibody staining
Fluorescent antibody staining
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Cell function prediction
Cell function prediction
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Tissue preparation
Tissue preparation
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Hematoxylin and eosin (H&E)
Hematoxylin and eosin (H&E)
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Hematoxylin
Hematoxylin
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Eosin
Eosin
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Common histological stains
Common histological stains
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Plasma membrane
Plasma membrane
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Nucleus
Nucleus
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Ribosomes
Ribosomes
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Endoplasmic reticulum (ER)
Endoplasmic reticulum (ER)
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Golgi apparatus
Golgi apparatus
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Smooth ER
Smooth ER
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Rough ER
Rough ER
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Secretory Vesicles
Secretory Vesicles
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Acidophilic Structures
Acidophilic Structures
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Mitochondria
Mitochondria
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Secreted Proteins
Secreted Proteins
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Cytoskeleton
Cytoskeleton
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Microtubules
Microtubules
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Intermediate filaments
Intermediate filaments
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Microfilaments (Actin filaments)
Microfilaments (Actin filaments)
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Visualization of Cytoskeleton
Visualization of Cytoskeleton
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Cell Structure Variation
Cell Structure Variation
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Cell Inclusions
Cell Inclusions
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Basophilic Cytoplasm
Basophilic Cytoplasm
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Study Notes
Cellular Structures - Learning Objectives
- The students will list and describe staining methods.
- Students will choose appropriate stains based on the tissue and desired outcome.
- Key cellular structures' functions will be described.
- Students will identify cellular structures via H&E staining appearance.
- Students will predict the prominence and function of cellular structures based on their function, or predict a cell function from given structures.
Review
- Students will be divided into groups of 2-3 for review activities at the beginning of class.
- Each group will discuss review questions, and the instructor will call on groups to share their answers.
Microscopy and Slide Preparation
- Microscopy is vital for histology study.
- Different microscopes are needed for different tasks:
- Traditional H&E staining requires a specific microscope.
- Fluorescent antibody (immunostaining) uses a unique microscopy technique.
- Examining very small structures (like ribosomes or chromosomes) needs a specialized electron microscope.
- Tissue samples are preserved either chemically (like formaldehyde) or by freezing in liquid nitrogen.
- Then, tissue samples are sectioned, mounted onto slides, and stained.
H&E Staining
- Hematoxylin and eosin (H&E) staining is a common histological technique.
- H&E is good for viewing general cell and tissue characteristics.
- Hematoxylin (basic dye) stains DNA, RNA, and ribosomes a purple color.
- Eosin (acidic dye) stains lysosomes, mitochondria, and collagen a pink color.
Common Histological Stains
- H&E is a general staining technique.
- Verhoeff's stain is used specifically for elastin fibers (black).
- PAS (periodic acid-Schiff) staining is for sugars and starches (purple/magenta).
- Mallory trichrome differentiates connective tissue (collagen - blue; elastin - pink/yellow; muscle - red).
- Silver stain is sensitive to small protein fibers.
Review: H&E Staining and Tissue Analysis
- If tissue cells appear bright pink after H&E staining, eosin is responsible for this color.
- The cell structures enriched by the appearance of staining can be identified. (e.g. presence of eosin in cell structures indicates possible abundance of mitochondria or secretory granules)
Cellular Structures and Diagrams
- Cells contain many organelles and subcellular structures that support functions. Various diagrams illustrate these structures and their relationships.
Cellular Structures to Review
- Important cellular components to study.
- Plasma membrane
- Nucleus
- Ribosomes
- Endoplasmic Reticulum- smooth ER and rough ER
- Golgi Apparatus
- Secretory vesicles
- Mitochondria
- Cytoskeleton
- Cellular inclusions (e.g. glycogen, lipid droplets)
Organizing into Small Groups to Review Cellular Functions
- Groups will brainstorm functions of structures assigned by the instructor (e.g. plasma membrane, nucleus, ribosomes, Rough ER etc).
Plasma Membrane
- The plasma membrane usually stains poorly with H&E (lipids stain poorly).
- A thin pink line (potential visualization) might be visible.
- Electron microscopy (EM) provides greater detail of plasma membrane structure.
The Nucleus
- The nucleus often stains basophilic (purple) due to the abundance of nucleic acids (DNA and RNA).
- Nuclei appearance can change, often relating to cell activity. -Examples of nucleus appearance changes are euchromatic nuclei.
Ribosomes
- Ribosomes, when concentrated, make the cytoplasm basophilic (purple).
- An abundant supply of ribosomes suggests increased protein synthesis activity.
Endoplasmic Reticulum
- Smooth ER: does not typically stain intensely with H&E due to lipid content.
- Rough ER: stains basophilic (purple) due to ribosomes.
Golgi Apparatus
- In most cell types, the Golgi is not readily apparent.
- In cells that produce and secrete proteins, the Golgi is more prominent, appearing as a pale pink area near the nucleus in H&E staining.
Secretory Vesicles
- In secretory cells, vesicles accumulate, sometimes being visible in H&E staining.
- The appearance of secretory vesicles differs based on the secretory product.
Mitochondria
- Mitochondria are acidophilic (stain pink) in H&E staining.
- Abundant mitochondria in a cell suggest high metabolic activity.
The Cytoskeleton
- A network composed of protein filaments throughout the cell.
- Provides structural support and assists in specialized functions (transport, movement, division).
- Three types of cytoskeletal elements: Microtubules, Intermediate Filaments, Microfilaments (actin filaments)
- Need techniques like electron microscopy or immunostaining to visualize cytoskeletal components
Cellular Inclusions
- Some cells have storage functions and contain cytoplasmic inclusions, which are not membrane-bound.
- Lipid droplets are examples of inclusions that do not typically stain with H&E.
- Glycogen inclusions can be stained with PAS (periodic acid-Schiff).
Cellular Structures: Structure = Function
- Diverse cell shapes and structures reflect the specialization for distinct functions.
- Examples include erythroblast (limited organelles), eosinophilic leukocyte (RER, Golgi, secretory granules), plasma cell (RER, Golgi), and pancreatic acinar cell (polarized RER, secretory granules).
Cell Appearance in H&E Staining
- Cell appearance in H&E varies according to structure and function.
- The detailed analysis of basophilic cytoplasm and euchromatic nuclei appearance would help in deducing cell structure, organelles and further inferring its function.
- Particularly the presence/absence of a particular organelles might also be helpful.
- Analyzing multiple structures within a cell can infer probable function of the cell.
Reminders
- No lab tomorrow.
- Lab on Friday, will discuss epithelial tissues.
- Quiz 1 will cover material from Monday and Friday, excluding the epithelium. It will be available on Canvas from Friday morning to Sunday evening.
- No class on Monday (Martin Luther King Jr. Day).
- Lab will resume on Thursday.
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Description
Test your knowledge on common histological stains and microscopy techniques in this quiz. Questions cover H&E staining, elastic fiber distribution, and immunostaining methods. Perfect for students studying histology or those in the biomedical field.
