GEN HISTOLOGY LABORATORY 1 & 2.pdf

GENERAL HISTOLOGY AND EMBRYOLOGY LABORATORY 1ST SEMESTER – SATURDAY BASIC MICROSCOPY ON / OFF SWITCH Turns the illuminator on or off MICROSCOPE...

GENERAL HISTOLOGY AND EMBRYOLOGY LABORATORY 1ST SEMESTER – SATURDAY BASIC MICROSCOPY ON / OFF SWITCH Turns the illuminator on or off MICROSCOPE BRIGHTNESS ADJUSTMENT “Mikros” (greek) “small” “Skopein” (greek) “to look” An instrument used to see objects Adjusts the intensity of light emitted that are too small for the naked eye by the illuminator Purpose MICROSCOPIC ILLUMINATOR To magnify a small object or to magnify the fine details of a larger Light source of the microscope object in order to examine minute Captures light from an external specimens that cannot be seen by source of a low voltage at about 100v the naked eye BASE COMPOUND BRIGHTFIELD MICROSCOPE Acts as the support of the microscope Carries the microscopic illuminators HEAD Acts as the support of the microscope Carries the microscopic illuminators ARM PARTS Connects the head and the base Supports the head of the microscope Used when carrying the instrument EYEPIECE Also known as the “oculars” which is part used to look through the microscope Found at the top PAGE 1 SABIDO, MARC KARYLLE S. – DMD II - COTTON PLIERS  GENERAL HISTOLOGY AND EMBRYOLOGY LABORATORY 1ST SEMESTER – SATURDAY Standard magnification = 10x (can Allows the control of the slides by have 5x-30x) using the mechanical knobs instead of moving it manually DIOPTER ADJUSTMENT LENSE Means to change focus on one eyepiece so as to correct for any Inverts the image of the object difference in vision between your inside the microscope. Inverted two eyes means that the image appears upside down and backward compared with the actual object This means that the slide must be moved in the opposite direction that you want the image to move. ○ Ex. if you move the slide to the right on stage, as you look through the microscope the NOSEPIECE OR TURRET slide appears to move left. “Revolving nosepiece” APERTURE Holds the objective lenses Movable hence it can revolve the objective lens as you change the Hole on the microscope stage magnification power through which the transmitted light form the light source reaches the stage OBJECTIVE LENSES CONDENSER Major lenses used for specimen visualization Magnification power of 4x - 100x Used to collect and focus light form the illuminator into the specimen Plays a major role in ensuring clear STAGE sharp images that are produced with a high magnification of 400x and Where the specimen is placed for above viewing Found under the stage next to the Contains ‘stage clips’ that hold the diaphragm specimen slides in place PAGE SABIDO, MARC KARYLLE S. – DMD II - COTTON PLIERS 2  GENERAL HISTOLOGY AND EMBRYOLOGY LABORATORY 1ST SEMESTER – SATURDAY DIAPHRAGM FIELD OF VIEW Also known as the ‘iris’ - The area a person can view through Controls the amount of light that a microscope reaches the specimen - Represented by a circle Adjustable apparatus hence the intensity and size of the beam of light can be adjusted Found under the stage TERMS MAGNIFICATION RELATIONSHIP - Ability to produce an image of an object at a larger scale than its actual size - As magnification increases, field of view decreases RESOLUTION/RESOLVING POWER - As you switch from scanning (40x) to high powers of magnification, we cannot see as much of the image. - Ability to distinguish between two closely spaced points on a specimen; smallest distance between two TOTAL MAGNIFICATION POWER structures at which they can be seen as separate objects - Light microscope: 0.2 um - This is the final magnified view of the image brought about by the product of the magnification power of the DEPTH OF FIELD oculars and the objective lens used - Objective lens Magnification x - The distance through which you can Ocular lens magnification = TMP move the specimen and still have it in focus PAGE SABIDO, MARC KARYLLE S. – DMD II - COTTON PLIERS 3  GENERAL HISTOLOGY AND EMBRYOLOGY LABORATORY 1ST SEMESTER – SATURDAY oriented and locate an area of HOW DOES A MICROSCOPE WORK? interest 6. Switch to a higher power to examine This straight-line path is disturbed by any field views more closely material of different refractive index. 7. To change magnification, simply rotate the nosepiece to bring one of Condenser (1.515) -> Air (1.00) -> Slide (1.515) the objectives into the light path; -> Mounting media (1.515) -> Cover slip you will hear/feel a “click” if the (1.515) -> Air (1.00) -> Objective (1.515) objective is in place 8. Turn off the light source Adding immersion oil (1.515) fills in the air 9. Center the mechanical stage gaps! 10. Position the nosepiece at the Scanner objective 11. Remove the slide from the stage 12. Clean the stage and lenses, wipe off any oil 13. Wrap the cord around/inside the arm 14. Carefully carry with two hands and gently place the microscope in the proper cabinet HANDLING RULES 1. Always carry the microscope upright (otherwise the eyepieces may fall out) 2. Use both hands, one grasping the arm or back slot and the other supporting the base 3. Position it on a flat and solid surface where it will not be easily knocked off TROUBLESHOOTING 4. Lenses must be clean for resolution. Only use lens paper or clean tissue. Do not remove any parts for cleaning Occasionally you may have trouble with 5. Always start with the lowest power working our microscope. Here are some objective (Scanner – 4x) to get common problems and solutions. 1. Image is too dark! PAGE SABIDO, MARC KARYLLE S. – DMD II - COTTON PLIERS 4  GENERAL HISTOLOGY AND EMBRYOLOGY LABORATORY 1ST SEMESTER – SATURDAY - Adjust the diaphragm, make 6. The condenser is too low or the iris is sure your light is on. closed 2. There's a spot in my viewing field, even when I move the slide the spot stays in the same place! PROBLEM DESCRIPTION: MICROSCOPE DOES NOT STAY IN - Your lens is dirty. Use lens FOCUS paper, and only lens paper to carefully clean the objective and ocular lens. The ocular COMMON ISSUE: lens can be removed to clean the inside. The spot is 1. The slide is not lying flat on the stage probably a spec of dust. 2. The tension adjustment needs to be 3. I can't see anything under high tightened power! - Remember the steps, if you 3. The nosepiece is not full engaged can't focus under scanning and then low power, you won't PROBLEM DESCRIPTION: DIRT / DUST be able to focus anything IN FIELD OF VIEW under high power. Start at scanning and walk through COMMON ISSUE: the steps again. 4. Only half of my viewing field is lit, it looks like there's a half-moon in 1. If debris moves when you rotate the there! eyepiece, then the dirt is on the - You probably don't have your eyepiece objective fully clicked into 2. If there is a diminished view under place one objective, then that objective has debris on it PROBLEM DESCRIPTION: 3. The top of the condenser or MICROSCOPE OUT OF FOCUS / POOR IMAGE QUALITY lamphouse has debris on it COMMON ISSUE: PROBLEM DESCRIPTION: SEEING DOUBLE IMAGE / HEADACHE WHILE USING MICROSCOPE 1. The slide is upside down 2. The stage height stop is set too low COMMON ISSUE: 3. The covership is missing or is too thick/thin 1. Microscope could be out of 4. A dry objective has oil on it - alignment if bumped or dropped common with 40x (clean of the oil) and needs repair 5. No oil or insufficient oil contact 2. The focusing eyepiece is all the way when using 100x objective up or down (make sure it is set at zero position) PAGE SABIDO, MARC KARYLLE S. – DMD II - COTTON PLIERS 5  GENERAL HISTOLOGY AND EMBRYOLOGY LABORATORY 1ST SEMESTER – SATURDAY - Forms a protective layer (Epidermis EPITHELIUM of the skin) - Absorption of substances (Intestines) - Secretion of substances (Intestines; glands) - Excretion of substances (Kidneys) - Transport/Exchange of substance (Lungs; Capillaries) BASIC TYPES OF TISSUES TISSUE CELLS EXTRACELLULAR MAIN MATRIX FUNCTIONS NERVOUS Elongated Very small Transmission cells with amount of nerve extremely impulses fine processes EPITHELIAL Aggregated Small Lining of polyhedral amount surface or cells body cavities; glandular secretion MUSCLE Elongated Moderate Strong contractile amount contraction; CELL POLARITY cells body movements CONNECTIVE Several types Abundant Support and - Asymmetric distribution of proteins, of fixed and amount protection of organelles, or cytoskeleton within wandering tissues/organ cells s groups of cells for them to carry out their intended functions EPITHELIUM 1. Apical Domain 2. Lateral Domain 3. Basal Domain - Layer or sheet of cells that covers a surface of lines in a cavity. - Avascular - Basement membrane separates the epithelial layer from the connective tissue 2 Functional Types: 1. Lining Epithelium 2. Glandular Epithelium FUNCTIONS PAGE SABIDO, MARC KARYLLE S. – DMD II - COTTON PLIERS 6  GENERAL HISTOLOGY AND EMBRYOLOGY LABORATORY 1ST SEMESTER – SATURDAY APICAL DOMAIN - Area directed towards the exterior surface or the lumen of an enclosed cavity or tube - May contain Special Surface Modifications APICAL DOMAIN: CILIA - Long, highly motile apical structures - Larger than microvilli - Abundant on cuboidal or columnar cells APICAL DOMAIN: MICROVILLI Function: - Move a current of fluid/suspended matter in one direction along the - Apical cytoplasmic projections epithelium - Uniform in length - Found in cells lining the small intestine - “brush/striated border” Function: - Increase surface area and allow absorption of substances PAGE SABIDO, MARC KARYLLE S. – DMD II - COTTON PLIERS 7  GENERAL HISTOLOGY AND EMBRYOLOGY LABORATORY 1ST SEMESTER – SATURDAY APICAL DOMAIN: STEREOCILIA - Long, nonmotile apical projections - Less common - Seen in the lining of the male reproductive system and inner ear sensory cells Function: - Increase surface area - Facilitate absorption - Motion-detecting function BASAL DOMAIN - Area directed towards the underlying connective tissue BASAL DOMAIN: BASEMENT MEMBRANE - Thin extracellular sheet of macromolecules Function: - Structural support - Anchoring of epithelium to the connective tissue - Scaffold for epithelial repair and regeneration - Mediates cell-cell interactions and LATERAL DOMAIN cellular migration - Area directed towards the adjacent cells - Allows communication between adjacent cells through specialized attachment areas PAGE SABIDO, MARC KARYLLE S. – DMD II - COTTON PLIERS 8  GENERAL HISTOLOGY AND EMBRYOLOGY LABORATORY 1ST SEMESTER – SATURDAY LINING EPITHELIUM How to classify? SIMPLE CUBOIDAL EPITHELIUM Cell Morphology Number of cell layers Simple - one cell layer - Single layer of cube-shaped cells Stratified - two or more layers Location: - Lining of kidney tubules - Ducts in some glands - Thyroid - Covering of the ovary Function: - Covering - Secretion SIMPLE SQUAMOUS EPITHELIUM - Single layer of flattened cells Location: - Lining of blood vessels (Endothelium) - Lining of cavities (Mesothelium) - Alveoli in the lungs Function: - Facilitates passage of substances (Transcytosis) SIMPLE COLUMNAR EPITHELIUM - Facilitates movement of the viscera - Secretion of some active molecules - Single layer of tall, rectangular cells - May contain specialized apical structures PAGE SABIDO, MARC KARYLLE S. – DMD II - COTTON PLIERS 9  GENERAL HISTOLOGY AND EMBRYOLOGY LABORATORY 1ST SEMESTER – SATURDAY Location: - Single layer only - Lining of stomach and intestine - Excretory ducts in some glands Location: - Gallbladder - Lining of the respiratory - Upper female reproductive tract - Passages - Epididymis Function: - Vas deferens - Protection - Lubrication Function: - Absorption - Protection - Secretion - Secretion - Propagation - Conduit for particle movement PSEUDOSTRATIFIED CILIATED COLUMNAR EPITHELIUM PSEUDOSTRATIFIED COLUMNAR EPITHELIUM - May appear as many layers because of the different levels of the nuclei; - But not all cells reach the free surface; all rest on the basement membrane PAGE SABIDO, MARC KARYLLE S. – DMD II - COTTON PLIERS 10  GENERAL HISTOLOGY AND EMBRYOLOGY LABORATORY 1ST SEMESTER – SATURDAY STRATIFIED SQUAMOUS EPITHELIUM STRATIFIED SQUAMOUS EPITHELIUM KERATINIZED - Multiple layers of flattened cells - Cells near the basement membrane are moved progressively toward the skin surface, becoming thin, inactive and lacking nuclei - May be Keratinized : - Epidermis of the skin Non-keratinized : - Covers the moist internal cavities (mouth, esophagus, vagina, STRATIFIED SQUAMOUS EPITHELIUM lower anal canal) NON KERATINIZED Function: - Protection - Secretion - Prevention of water loss STRATIFIED CUBOIDAL AND COLUMNAR EPITHELIUM - Multiple layers of square-shaped or rectangular cells - Relatively rare Location: - Salivary and sweat glands Function: - Protection - Secretion PAGE SABIDO, MARC KARYLLE S. – DMD II - COTTON PLIERS 11  GENERAL HISTOLOGY AND EMBRYOLOGY LABORATORY 1ST SEMESTER – SATURDAY TRANSITIONAL EPITHELIUM - Multiple layers of square-shaped to flattened cells with a superficial layer of large, dome-like cells called “umbrella cells” - Also termed as “UROTHELIUM” STRATIFIED CUBOIDAL EPITHELIUM Location: - Lining of the urinary tract (renal calyces, ureters, bladder) Function: - Architecture allows it to DISTEND, accommodating fluid accumulation STRATIFIED COLUMNAR EPITHELIUM PAGE SABIDO, MARC KARYLLE S. – DMD II - COTTON PLIERS 12