Prokaryotic Cell
Prokaryotic Cell Structure
Colony Morphology: Characteristics such as texture, margin, elevation, and shape which help identify bacterial colonies.
Shape: Circular, irregular, filamentous, rhizoid.
Margin: Entire, undulate, lobate, curied.
Elevation: Flat, raised, convex, pulvinate, umbonate.
Size: Measured in millimeters (mm), described as pinpoint, small, medium, large.
Texture: Dry, moist, mucoid, brittle, viscid, viscous, butyrous.
Opacity: Transparent, opaque, translucent, iridescent.
Odour: Specific smells aiding in identification, e.g., earthy odour from actinomycetes, fruity smell from some fungi, faecal odour from E. coli.
Colour: Pigmented colonies seen in culturing.
Prokaryotic Genetic Material
Structure: Prokaryotic cells typically contain a single circular DNA molecule known as the prokaryotic chromosome.
Properties:
Circular or linear.
Contains extrachromosomal DNA (plasmids).
Single copy per gene, generally less repetitive DNA.
Ribosomes
Tiny particles composed of protein and RNA.
Functions as the site for protein synthesis, with numbers varying by growth stage (up to 20,000 in rapidly growing E. coli).
Prokaryotic ribosomes are classified as 70S, smaller than eukaryotic (80S) ribosomes.
Structures Surrounding the Bacterial Cell
Capsule: Protects against phagocytosis, enhances adherence.
Slime Layer: Less structured than capsules, provides similar functions.
Pili: Hairlike projections for attachment to surfaces, some for genetic transfer (sex pili).
Flagella: Motility structures made of flagellin, aiding in movement.
Mesosome: Extensions of the cell membrane involved in various cellular functions.
Cell Wall and Peptidoglycan Structure
Function: Provides structural support and shape, protects against osmotic lysis, and can contribute to pathogenicity.
Peptidoglycan Composition: Made of repeating N-acetylglucosamine (NAG) and N-acetylmuramic acid (NAM) units, linked by peptide chains.
Gram-Positive vs. Gram-Negative:
Gram-positive: Thick peptidoglycan layer (90%), teichoic acids present.
Gram-negative: Thin peptidoglycan layer (5-10%), outer membrane with lipopolysaccharide (LPS) and periplasmic space.
Staining Techniques
Types:
Negative Staining: To observe shape.
Simple Stain: Uses a single dye for overall visualization.
Differential Stains: E.g., Gram and acid-fast staining, allow distinguishing between types of bacteria based on cell wall composition.
Special Stains: For specific structures such as flags, capsules, and spores.
Nuclear Stain: Acridine orange helps visualize DNA.
Procedures for Specific Staining Techniques
Gram Staining Protocol:
Primary stain (crystal violet) applied.
Mordant (iodine).
Decolorization with alcohol.
Counterstain (safranin).
Acid-Fast Staining: Identify mycobacteria, involves carbol fuchsin and acid alcohol.
Capsule Staining: Uses nigrosin and safranin to visualize capsules as clear halos against a dark background.
Spore Staining Protocol: Malachite green used for spores, water for decolorizing, and safranin for vegetative cells.
Summary of Key Structures and Functions in Prokaryotic Cells
Common Structures: Plasma membrane, cytoplasm, ribosomes, nucleoid, cell wall, capsule, and others such as flagella and pili for attachment.
Functions: Protect against external factors, aid in nutrient transport, enable motility, and facilitate reproduction and genetic diversity through plasmids and conjugation.