Intro to Histology and Bacterial Shapes
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Questions and Answers

What is the primary goal of histology?

  • To develop new staining methods
  • To analyze chemical compositions of cells
  • To study macroscopic structures
  • To identify correlations between structure and function (correct)

Hematoxylin and Eosin (H&E) staining is primarily used for immediate observational feedback in tissue analysis.

True (A)

What are the typical thicknesses of cross-sectional slices prepared for histological observation?

2-7 μm

Before observing tissue samples, they must be __________ to prevent decay.

<p>chemically fixed</p> Signup and view all the answers

Match the histological techniques with their purposes:

<p>Microtome = Slices hardened resin into thin sections H&amp;E staining = Differentiates cellular structures in tissue Cryostat = Prepares frozen sections of tissue Chemical fixation = Prevents decay of samples</p> Signup and view all the answers

Which of the following describes the appearance of tissues once cross-sectional slices are prepared?

<p>Colorless and transparent (D)</p> Signup and view all the answers

Histology only applies to isolated tissues, not cultured cells.

<p>False (B)</p> Signup and view all the answers

What is the purpose of applying stains to cells in histology?

<p>To enhance contrast and permit visualization</p> Signup and view all the answers

Which arrangement involves cocci in pairs?

<p>Diplococcus (A)</p> Signup and view all the answers

Bacillus bacteria can only exist as single cells.

<p>False (B)</p> Signup and view all the answers

What is the diameter range of coccus-shaped bacteria?

<p>0.5-1.0 μm</p> Signup and view all the answers

Bacteria commonly reproduce by __________.

<p>binary fission</p> Signup and view all the answers

Match the bacterial shapes to their descriptions:

<p>Coccus = Spherical or oval shape Bacillus = Rod-shaped Vibrio = Comma-shaped Spirillum = Thick and rigid spiral</p> Signup and view all the answers

Which type of spiral-shaped bacteria is thin and flexible?

<p>Spirochete (D)</p> Signup and view all the answers

Plant cells can be found in sizes ranging from 10-100 μm long.

<p>True (A)</p> Signup and view all the answers

Name one distinguishing feature of plant cells that is not found in mammalian cells.

<p>Cell wall</p> Signup and view all the answers

Which adjustment knob should be used to bring a specimen into initial focus when using the 4x objective lens?

<p>Coarse adjustment knob (D)</p> Signup and view all the answers

Coarse adjustment can be used with any objective lens without risk of damage.

<p>False (B)</p> Signup and view all the answers

What should be observed during turret rotation when increasing magnification?

<p>The distance between the objective lens and the slide</p> Signup and view all the answers

Before using the 100x objective lens, special ______ is required on the slide.

<p>oil</p> Signup and view all the answers

Match the following objective lenses with their descriptions:

<p>4x = Scanning objective lens 10x = Medium objective lens 100x = Oil immersion objective lens 40x = Higher magnification objective lens</p> Signup and view all the answers

What is the measurement distance between each line on a stage micrometer?

<p>10μm (A)</p> Signup and view all the answers

An ocular micrometer has specific measurements assigned to its lines.

<p>False (B)</p> Signup and view all the answers

What type of ruler is used to calibrate a microscope if a stage micrometer is not available?

<p>Regular ruler</p> Signup and view all the answers

Which part of the document focuses on the observation of tissue samples?

<p>Part B: Tissue observation (B)</p> Signup and view all the answers

Table 1.4 includes information about the observation of mammalian cells.

<p>False (B)</p> Signup and view all the answers

What is the purpose of the staining procedure in tissue observation?

<p>To enhance visibility of cellular structures under a microscope.</p> Signup and view all the answers

The subcellular structure commonly stained with DAPI is the ______.

<p>nucleus</p> Signup and view all the answers

Match the following staining procedures with their target:

<p>H&amp;E = General tissue structure DAPI = Nucleus Gram Stain = Bacterial classification Giemsa = Blood cell morphology</p> Signup and view all the answers

Which objective lens would typically provide a higher magnification for observing bacterial samples?

<p>100x (D)</p> Signup and view all the answers

Part C includes wet mount preparation techniques.

<p>False (B)</p> Signup and view all the answers

What information is provided in the 'FOV size' section of the tables?

<p>The size of the field of view under the microscope.</p> Signup and view all the answers

What is the purpose of calibrating the ocular micrometer?

<p>To calculate the real size of the viewed specimen (D)</p> Signup and view all the answers

The stage micrometer is used to adjust the focus of the microscope.

<p>False (B)</p> Signup and view all the answers

What is the length of each division on the stage micrometer?

<p>10 μm</p> Signup and view all the answers

The ocular micrometer is divided into _____ equivalent units.

<p>100</p> Signup and view all the answers

Match the following components with their corresponding properties:

<p>Stage Micrometer = 1 mm long with 100 divisions Ocular Micrometer = Divided into 100 units with unknown distance 35 mark on Ocular = Corresponds to 30 divisions on Stage Each division on The Stage = 10 μm apart</p> Signup and view all the answers

How is the distance between the lines of the ocular micrometer determined?

<p>By superimposing with the stage micrometer (A)</p> Signup and view all the answers

The point of overlap between the stage micrometer and ocular micrometer is where the 600 μm mark is located.

<p>False (B)</p> Signup and view all the answers

How many divisions correspond to 300 μm on the ocular micrometer?

<p>35 divisions</p> Signup and view all the answers

Which of the following is NOT a rule for labelling a scientific drawing?

<p>Cross lines when labelling for clarity. (B)</p> Signup and view all the answers

A dry mount slide preparation uses water to suspend the sample.

<p>False (B)</p> Signup and view all the answers

What is the maximum amount of liquid medium recommended for a wet mount slide preparation?

<p>One drop</p> Signup and view all the answers

In wet mount preparation, the coverslip should be placed at a _______ degree angle.

<p>45</p> Signup and view all the answers

Match the slide preparation method with its characteristic:

<p>Wet mount = Uses liquid medium for observation Dry mount = Does not use water or liquid Coverslip method = Used to cover the specimen Suspension solution = Liquid added to aid visibility</p> Signup and view all the answers

Which step is first in preparing a wet mount slide?

<p>Place the sample specimen in the middle of a clean slide. (D)</p> Signup and view all the answers

Labels on a scientific drawing can be drawn in any direction.

<p>False (B)</p> Signup and view all the answers

What is the potential consequence of excessive use of suspension solution in wet mount preparation?

<p>Specimen may run off the edges of the slide.</p> Signup and view all the answers

Flashcards

Histology

The practice of using stains to enhance the visibility of cell and tissue characteristics under a microscope.

Histology - Broad Definition

The study of the minute structure of cells, tissues, and organs using magnification to understand how structure relates to function.

Chemical Fixation

The process of preserving biological samples by using chemicals to prevent decay.

Embedding

The process of embedding biological samples in a solid medium, like wax, for easy slicing.

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Microtome

A device used to cut thin slices of tissue embedded in wax or resin.

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H&E Stain

A staining technique that uses hematoxylin (blue) and eosin (pink) to differentiate structures within cells and tissues.

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Contrast

The ability to distinguish between different structures within a sample due to differences in color or appearance.

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Staining Methods

The use of dyes or stains to enhance the contrast of cellular and tissue components, making them easier to see.

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Coarse Adjustment Knob

The knob used to quickly adjust the stage and bring the specimen into focus. It should only be used with the scanning or 4x objective lens.

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Fine Adjustment Knob

The knob used to finely adjust the stage and bring the image into sharp focus. It's used with all objective lenses.

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Turret

The rotating part of the microscope that holds and switches between different objective lenses, each offering different magnification. Important to note that moving it requires checking the distance between the lens and slide for clearance.

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Field of View (FOV)

The diameter of the field of view (FOV) in a microscope. It is the circular area visible through the objective lens.

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Staining

A technique used to highlight specific structures within a cell or tissue sample by selectively staining them with dye.

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Hematoxylin and Eosin (H&E) Staining

Hematoxylin and eosin (H&E) staining is a common staining technique used in histology. Hematoxylin stains nuclei blue, while eosin stains the cytoplasm and other tissue components pink.

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Resolution

The ability of a microscope to distinguish between two closely spaced objects as separate entities. Higher resolution means greater detail is visible.

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Wet Mount Preparation

A technique for preparing slides involving placing a sample on a slide and covering it with a coverslip. It allows observation of living organisms and unstained specimens.

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Magnification

The degree to which an object appears larger under a microscope. Magnification is calculated by multiplying the magnification of the objective lens by the magnification of the eyepiece.

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Microscopy of Microorganisms

Microscopic observation of microorganisms and their cellular features. Provides information about the shape, size, and arrangement of bacteria.

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Microscopy of Tissue Samples

Microscopic observation of tissue samples. Helps to identify the different types of tissues and their structural components.

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Coccus bacteria

Spherical or oval shaped bacteria, typically 0.5-1.0μm in diameter. Can exist in various arrangements, including pairs (diplococcus), chains (streptococcus), tetrads (square of four), sarcina (cube of eight), and staphylococcus (irregular clusters).

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Bacillus bacteria

Rod-shaped bacteria, usually 0.5-1.0μm wide and 1-4μm long. Can be found as single bacilli, pairs (diplobacillus), chains (streptobacillus), or oval-shaped coccobacillus.

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Spiral bacteria

Bacteria with a spiral shape. They can be: Vibrio (incomplete spiral, comma-shaped), Spirillum (thick, rigid spiral), or Spirochete (thin, flexible spiral, can be very long).

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Plant cells

Cells that belong to the Eukarya Domain and Plantae kingdom. Plant cells share features with mammalian cells like nuclei, but also have unique characteristics such as cell walls, chloroplasts, and vacuoles. They can range in size from 10-100 μm long.

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Mammalian cells

Cells that belong to the Eukarya Domain. Mammalian cells contain nuclei, mitochondria, lysosomes, endoplasmic reticulum, and other organelles. They are specialized to perform specific functions in different organs and tissues.

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Binary fission

A form of asexual reproduction where a single parent cell divides, giving rise to two identical daughter cells. Common in bacteria.

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Bacterial spores

Resistant structures formed by some bacteria, allowing them to survive harsh conditions. They are basically a protective coat around the bacterial DNA.

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Bacterial flagella

Whip-like structures used by some bacteria for movement. They rotate like propellers.

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Wet Mount

A type of slide preparation where the sample is suspended in a liquid medium, such as water or saline solution.

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Dry Mount

A type of slide preparation where the sample is placed directly on the slide without any liquid medium.

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Coverslip

A small, transparent piece of glass that is placed over the sample on a microscope slide to protect it and flatten it.

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Suspension Solution

The liquid medium used to suspend the sample in a wet mount slide preparation.

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Slide Preparation

The process of placing a sample on a microscope slide for observation. It can be performed using a wet mount or dry mount method.

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Placing the Specimen

The act of placing the specimen on a slide, ideally in the center.

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Placing One Drop of Suspension Solution

A single drop of liquid medium placed on top of the slide to suspend the specimen.

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Lowering the Coverslip

The process of carefully lowering the coverslip onto the specimen to ensure it is completely covered and free of air bubbles.

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What is ocular micrometer calibration?

The process of measuring the exact distance between lines on an ocular micrometer by aligning it with a stage micrometer.

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What is a stage micrometer?

A specialized ruler used in microscopy to measure the size of objects viewed under the microscope.

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What is an ocular micrometer?

A specialized ruler built into the microscope eyepiece, used to measure the size of objects viewed under the microscope.

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What is the key feature of a stage micrometer?

The distance between lines on a stage micrometer is a fixed value, usually 10 micrometers (µm) per division. This fixed value is used for calibration.

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What is the objective of ocular micrometer calibration?

The distance between each line on an ocular micrometer is unknown until calibration. Calibration allows you to determine the distance between lines on the ocular micrometer.

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How is ocular micrometer calibration performed?

By superimposing the stage micrometer on the ocular micrometer, we match the lines and determine the corresponding divisions. This allows us to calculate the actual distance on the ocular micrometer.

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What is the practical application of ocular micrometer calibration?

After calibration, the ocular micrometer can be used to measure the actual size of objects observed under the microscope by counting the divisions covered by the object.

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Why is ocular micrometer calibration important?

Calibration of the ocular micrometer is crucial for accurate measurement and analysis of specimens in microscopy.

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Study Notes

Laboratory 1 - Histology & Basic Microscopy

  • Rationale: This lab exercise observes various cell types (microbes to mammals) under a microscope. It also introduces staining techniques used in biomedical science. Students practice wet mount slide prep and compare cell structures.

Objectives

  • Objective 1: Practice oil immersion microscopy for viewing microorganisms.
  • Objective 2: Compare sizes and shapes of bacterial and mammalian cells.
  • Objective 3: Prepare and analyze plant cell wet mounts.
  • Objective 4: Understand tissue and cell staining for proper visualization.
  • Objective 5: Explain the H&E staining mechanism.
  • Objective 6: Perform H&E staining on cultured cells.

Introduction

  • Compound Microscope: A light microscope combines magnifying lenses and light to enlarge extremely small objects not visible to the naked eye.
  • Bacteria: Bacteria are unicellular prokaryotes distinguished by shape (coccus, bacillus, spiral) and structure. Most bacteria are 0.5-1.0 µm in diameter and 1-8 µm in length. They can be in different arrangements (pairs, chains, clusters, etc.).
  • Plant Cells: Plant cells are eukaryotic and have unique features like cell walls and chloroplasts. Their sizes vary widely, between 10-100 µm.
  • Mammalian Cells: Mammalian cells are eukaryotic and complex, with diverse organelles (nucleus, mitochondria, etc.). Their structures and organization vary depending on their function and specific tissue.

Histology

  • Histology: The study of the microanatomy of cells, tissues, and organs under magnification to understand cellular structure-function relationships.
  • H&E Staining: A common staining technique for general tissue identification in pathology and histology. Hematoxylin stains acidic structures (e.g., nuclei) purple/blue. Eosin stains basic structures (e.g., cytoplasm) pink/red. This contrast highlights differences in cell organization.

Oil Immersion Microscopy

  • Oil Immersion Microscopy: An advanced microscopy technique that increases the resolving power of the microscope by eliminating light distortion.
  • Procedure: Focus on the specimen, place immersion oil on the cover slip (on the specimen), and use the high power objective lens.

Experimental Procedure

  • Materials and Reagents: The lab lists materials like cells (C2C12, HEK293), ethanol, formaldehyde, H&E stain kit, PBS, etc.
  • Part A - Visualization of Microorganisms: A procedure for observing and documenting various bacterial types, their shapes, and arrangements at 1000x magnification using a microscope.
  • Part B - Tissue observation: Observing H&E-stained mammalian tissue slides under varying magnifications (4x, 10x, 40x, and 100x).
  • Part C - Cell culture staining: A procedure to stain cells, using a specific set of steps for fixation, permeabilization, and staining with hematoxylin and eosin.
  • Part D - Slide Preparation – Onion Epidermis: A procedure to prepare a wet-mount slide of onion epidermis for viewing under a microscope.

Data Collection and Post-Lab Assignment guidelines

  • Submission: Electronic submission, including observations and photos of various parts of the procedure, for each part. Includes tables to categorize data.

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Related Documents

BMED 2000 Laboratory 1 - Histology & Basic Microscopy PDF

Description

This quiz covers the fundamental concepts of histology and the various bacterial shapes and arrangements. Test your knowledge on staining techniques, tissue preparation, and the characteristics of different microorganisms. Ideal for students studying histology or microbiology.

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