Histopathologic & Cytologic Techniques Preliminary Lab PDF

HISTOPATHOLOGIC & CYTOLOGIC TECHNIQUES PRELIM LABORATORY | FIRST SEMESTER WEEK 1: LABORATORY SAFETY AND INSTRUMENTATION HISTOPATHOLOGY Art of analyzing and interpreting the shapes, sizes, and arch...

HISTOPATHOLOGIC & CYTOLOGIC TECHNIQUES PRELIM LABORATORY | FIRST SEMESTER WEEK 1: LABORATORY SAFETY AND INSTRUMENTATION HISTOPATHOLOGY Art of analyzing and interpreting the shapes, sizes, and architectural patterns of cells and tissues within a given specific clinical background. Activities Before testing proper Sample collection, transport, labeling, and accessioning Tissue processing Pre-analytical Submission of slide for reporting Any error can endanger the quality of histopathology test results or report Tissue or slide reading Analytical Preparation of report or test results Delivery of report or test results Post-analytical Archiving of test results Storing reported specimen Safe disposal of specimen RISK MANAGEMENT RISK MANAGEMENT Process of ensuring and maintaining personal as well as environmental health and safety in the laboratory. The first step is to identify all electrical, mechanical, and biological hazards that can potentially cause harm in the laboratory. o Look for standard operating procedure. Inventory of chemical reagents must be on hand and obsolete chemicals should be routinely disposed of. HISTOPATHOLOGIC & CYTOLOGIC TECHNIQUES PRELIM LABORATORY | FIRST SEMESTER WEEK 1: LABORATORY SAFETY AND INSTRUMENTATION TYPES OF HAZARDS HAZARD ADDITIONAL INFO EXAMPLES Inventory of chemical reagents must be on hand a Can be encountered thru usage or poor storage. The “lab standard” o applies to the laboratory use of chemicals o mandates written Standard Operating Procedures (SOPs)  address the particular hazards and precautions required for safe use. Labelling Every chemical should be labeled with certain basic information, including: 1. Chemical name  If it’s a mixture it should include all the names of the ingredients 2. Manufacturer’s name and address (if chemical is purchased commercially) o Reagents or solutions made within the day should include the name of the person who made it. EXAMPLES: 1. Cleaning agents and disinfectants, Chemical 3. Date purchase or date made 2. drugs, Hazards 3. anesthetic gases, 4. Expiration date (if known) 4. solvents, 5. Hazards warning and safety procedure 5. paints, 6. compressed gases DIFFERENT TYPES OF CHEMICALS:  Chemicals that can cause reversible inflammatory effects at the site of Irritants contact with living tissue, especially the skin, eyes, and respiratory passages Cause destruction or irreversible alterations when exposed to living tissue o Can destroy inanimate surfaces (metal) Corrosive o Can be corrosive to tissue but not to steel, and vice versa. o Few are corrosive to both tissue and steel Cause allergic reactions in some exposed workers, not just in Sensitizers hypersensitive individuals o It can occur at work because of high exposure level. HISTOPATHOLOGIC & CYTOLOGIC TECHNIQUES PRELIM LABORATORY | FIRST SEMESTER WEEK 1: LABORATORY SAFETY AND INSTRUMENTATION Substances that can induce tumors, not only in experimental animals but also to humans Include: 1. chloroforms, 2. chromic acid, Carcinogens 3. formaldehyde, 4. nickel chloride, 5. potassium dichromate, 6. carcinogenic dyes  auramine,  basic fuchsin,  any dye derived from benzidine (including Congo red and diamino-benzidine Capable of causing death by ingestion, skin contact, or inhalation at certain specified concentrations. Include: Toxic Material 1. methanol, 2. mercury, 3. chromic acid, 4. osmium tetroxide, 5. uranyl nitrate. Slips and falls from working in wet locations and the ergonomic hazards of lifting, pushing, pulling, and repetitive tasks Other physical hazards often unnoticed are : 1. electrical, Physical 2. mechanical, Hazards 3. thermal hazards o Ignoring these can have potentially serious consequences Many operations in the lab can result in lab workers assuming sustained or repetitive awkward postures such as looking at slides on microscope for extended periods o Don’t forget to practice proper posture when doing tasks Biological Anything that can cause diseases to humans regardless of the source. Include: Hazards 1. infection agents and their toxins Universal precaution: treat all specimens as infectious. HISTOPATHOLOGIC & CYTOLOGIC TECHNIQUES PRELIM LABORATORY | FIRST SEMESTER WEEK 1: LABORATORY SAFETY AND INSTRUMENTATION 2. contaminated solutions, specimens, or Allergens: one of the most important health hazards, yet they are frequently overlooked objects. Molds and fungi : are produced and release millions of spores small enough to be air, water, or insect-borne which may have negative effects on human health including allergic reactions, asthma, and other respiratory problems. HISTOPATHOLOGIC & CYTOLOGIC TECHNIQUES PRELIM LABORATORY | FIRST SEMESTER WEEK 1: LABORATORY SAFETY AND INSTRUMENTATION MICROSCOPE Main Framework Use and care of Microscope MICROSCOPE contains the light source, provide supports for the microscope.  an equipment that is used by both the pathologist Base o Should be large and solid and histotechnologist. o Pathologist: examines the slide under the enough to allow the microscope microscope to identify a disease process or an to stand on its own. abnormality that will directly affect the patient’s treatment. also supports the microscope. It o Histotechnologist: examines the same slide holds the magnifying and adjusting microscopically for quality control to determine system. whether all technical processes are done Arm o Can also be used as a handle, properly and if a slide of diagnostic quality has provided that one arm is holding been achieved. or carrying the arm and one The microscope must accomplish the following: hand is holding the base 1. It must magnify the object. 2. It must resolve the details of the object. platform where the slide is placed for 3. It must make these details visible. examination o Substage: located under the stage and holds the condenser factor by which an image appears Stage Magnification and diaphragm. to be enlarged. o Mechanical stage: permits the shortest distance between two movement of the stage while Resolution points that can still be visually holding the slide in the place of distinguished as separate focus ability of the microscope to Resolving distinguish between small objects Power that are close together. PARTS Parts of the Compound Microscope Huygenian: “mono” 1. Ocular (eye piece), 2. body or tube, 1 eye piece to view 3. coarse focusing knob, the specimen 4. objective lens, o DISADVANTAGE 5. movable stage, : the usage of an 6. condenser lenses, LCD camera is 7. field (iris diaphragm), Monocular restricted 8. mirror, because that 9. light. would already occupy the eye COMPOUND MICROSCOPE piece  has more than one lens, own light source (found on the base of the microscope) Viewing o ADVANTAGE: o usually use binocular eyepiece type of Heads inexpensive and microscope “Eye lightweight Piece”  OBJECTIVE LENSES Ramsden: “bi” 1. scanner, 2 eye pieces 2. ow power objective (LPO), o More convenient 3. high power objective (HPO), Binocular and comfortable to 4. oil immersion objective (OIO). use o What we used in the laboratory o The most common choice Compensating: “tri” Trinocular 3 eye pieces HISTOPATHOLOGIC & CYTOLOGIC TECHNIQUES PRELIM LABORATORY | FIRST SEMESTER WEEK 1: LABORATORY SAFETY AND INSTRUMENTATION o There is 3rd ▪ 10x objective × 10x eyepiece that can eyepiece = 100x be used for an LCD camera or for ▪ 40x objective × 10x simultaneous eyepiece = 400x viewing of another person o Most expensive Mounted on nose piece (which is located at the end of the body tube) It forms magnified real image Objectives is consisting of system of lenses located at the end of a turret. The purpose of those objective is to either increase or decrease the magnification. Magnification of objective = optical tube length / focal length o Normal optical tube length: 160 mm o Focal length: the distance between outer lens of objectives and the cover glass of the slide under examination. Objective Lenses Types: o Scanner (red): 4x o Low power (yellow): 10x o High power (blue): 40x o Oil Immersion (white): 100x Total magnification = objectives × eyepiece o Need to get the magnification of the eyepiece as well o Examples: (just multiply the magnification of the objective and eyepiece) ▪ 4x scanning objective × 10x eyepiece = 40x HISTOPATHOLOGIC & CYTOLOGIC TECHNIQUES PRELIM LABORATORY | FIRST SEMESTER WEEK 1: LABORATORY SAFETY AND INSTRUMENTATION Types of Microscopes MICROSCOPE ADDITIONAL INFO PURPOSE In some, compound microscope is also called as bright field microscope o because it is the one commonly used o However, bright field microscope is under the compound microscope. The term “bright field” is derived from the fact that the specimen is dark, and it is contrasted by the surrounding bright viewing field Generally used in compound microscopes, where light either passed Bright Field through or reflected off a specimen Microscope A specimen is placed on the stage of the microscope and incandescent light from the microscope’s light source is aimed at a lens beneath the specimen called as condenser. o Condenser: contains aperture diaphragm to help control and focus the light on the specimen ▪ After that, the light will pass through the specimen and is collected by the objective lenses situated just above the turret ▪ Then, the objectives will magnify the light and transmit it to the ocular lenses or eye piece up to the user’s eyes used to observe unstained and transparent samples causing them to be clearly visible and appear brightly lit against a dark, almost purely Also known as dark ground microscopy black background Dark Field It blocks central light with a condenser so only oblique rays will hit the Useful in demonstrating: Illumination object. 1. Treponema pallidum o The effect of that is that the specimen being viewed will appear 2. Leptospira, bright on a dark or black background 3. Campylobacter jejuni, 4. endospores A type of light microscopy Useful in studying cell cycle in live cells Phase It doesn’t require staining to view enhances contrast of transparent and colorless objects by influencing Contrast specimen the optical path of light Microscopy show components in a cell or bacteria which would be very difficult to see in an ordinary light of bright field microscope HISTOPATHOLOGIC & CYTOLOGIC TECHNIQUES PRELIM LABORATORY | FIRST SEMESTER WEEK 1: LABORATORY SAFETY AND INSTRUMENTATION enables visualization of internal cellular components and useful for the diagnosis of tumor cells o Living cells are being studied, as well as on how they proliferate through cell division For examination of growth and behavior of living cells in a cell culture 2 essential components: o Polarizer: below the specimen stage ▪ Usually fixed in the left to Contrast enhancing technique that improves the quality of the image right, east-west direction obtained with birefringent materials when compared to other techniques such as bright field microscopy, phase contrast Polarized ▪ Some are rotatable through microscopy, and dark field microscopy. Microscopy 360° Designed to examined specimens that are visible primarily due to their o Analyzer: above the objectives optically anisotropic character ▪ Aligned north-south Can be used to have a better view of more complex structures such as microtubules and overlapping structures. ▪ Some are rotatable through 360° ▪ Can be move in and out of the light path as required. In fluorescence microscopy, many wavelengths of light, ranging from the ultraviolet to the visible can be used to cause samples to fluoresce and allow viewing by eye or with the use of specifically sensitive camera When certain compounds are illuminated with high energy light, they emit light of a lower frequency. It refers to any microscope that uses fluorescence to generate image Fluorescence Microscope Specimen will be stained by a fluorescent dye. When you examined it under a microscope with ultraviolet light, they are seen as bright object against a dark background. Example: Auramine Rhodamine o Yellow: it is the tubercle bacilli o Acridine orange: gives orange red with RNA and yellow green with DNA immunofluorescence HISTOPATHOLOGIC & CYTOLOGIC TECHNIQUES PRELIM LABORATORY | FIRST SEMESTER WEEK 1: LABORATORY SAFETY AND INSTRUMENTATION Uses a beam of highly energetic electrons to examine objects on a very fine scale. It uses accelerated electrons as a source of illumination Wavelength of electron: up to 100,000x shorter than that of visible light photon Electron microscopes have high resolving power than your light microscope It creates an image of the sample’s internal structure It uses a broad beam of electron Transmission Electron It cost more than a scanning electron microscope. Microscopes More harmful and detrimental to human health (TEM) can reveal the structure of smaller objects. because of its higher energy electron beams This examination can yield the information It produces flat 2D images about morphology and composition Electron Ideal for looking for internal details of small sample at a Microscope near atomic resolution It produces 3D images It uses a fine beam of focus electron to scan the surface of the sample If there is a need to look at the large area and only Scanning need the surface details, SEM is ideal. Electron Microscopes SEM Images (SEM) HISTOPATHOLOGIC & CYTOLOGIC TECHNIQUES PRELIM LABORATORY | FIRST SEMESTER WEEK 1: LABORATORY SAFETY AND INSTRUMENTATION are used to assess the quality LABORATORY QUALITY MANAGEMENT SYSTEM of staining  There are stained Degree to which healthcare services preparations already in strive to provide accurate, precise records since we have set Quality desired outcomes for patients and are retention period consistent with current professional knowledge.  Somewhat like an internal quality checking where Freedom from accidental injury random picking of one prepared slide is done. Hazards that can be encountered: Then, the pathologist toxic chemicals, biohazards or available will be the one pathogenic microorganisms, who will read it while the Safety mechanical, electrical, and fire histotechnologist will give hazard or thermal hazards his/her report for quality control. After that, it will o It is important that you are be countercheck to the conscious about the safety actual result they practices within the laboratory randomly picked for stained preparations. System of routine technical activities o Distributive system: other It provides routine and consistent participating laboratories are checks to identify and address asked to stain sections that errors and omissions Quality have been submitted by the Control It ensures the integrity of data scheme organizer (which is being released usually the reference o The data being released are laboratory) correct and complete ▪ Will be given a sample or Records all quality control prepared slide that is not activities. yet stained. It can also be a tissue block and the one Planned system of review that will section it will be procedures conducted by the participating personnel not directly involved in laboratories. Afterwards, it the laboratory process. will be stain, mount, etc. Data of QC provides the data for QA ▪ The laboratories will send in their results and it will Quality assessment programs: be paralleled or correlated o College of American with the actual and known Pathologists (CAP) result of the scheme o United Kingdom National organizer External Quality Assessment Service (UK System is used to approach, NEQAS) evaluate, and identify Quality Assurance ▪ There is NEQAS available opportunities to improve quality in the Philippines before problem occur through Continuing evaluation of all systems or Quality Can correlate the errors, Improvement processes in the laboratory. complaints, failures, or other unexpected results if there will be Main Goal: to improve the any potential care or safety through recognition of potential problems or errors before they can occur. Two distinct systems: o Selective system: stained preparations from departmental archival records HISTOPATHOLOGIC & CYTOLOGIC TECHNIQUES PRELIM LECTURE | FIRST SEMESTER WEEK 2: INTRODUCTION TO GENERAL PATHOLOGY & HISTOTECHNIQUES PATHOLOGY Terms to Remember  From ancient Greek words: pathos & logos Pathos pain or suffering Logos study of Development of a disease or morbid Pathogenesis  Study of Diseases condition  Aka pathobiology  Physician who specializes in  Study of the structural & functional changes in cells, tissues, interpretation and who diagnoses and organs that underlie diseases. changes caused by diseases in the  Study of cellular abnormalities body.  Mainly bridges the discipline of basic science & clinical  Specialist or expert in determining the practice. origin and development of the  According to Rudolf Virchow, all diseases originated at a diseases and microscopic analysis of cellular level. body tissues.  Pathologist Diseases & underlying mechanisms are best understood  They study all the aspects of diseases in the context of normal cellular structure and function. with an emphasis on the nature,  A branch of medical science that involves the study and causes and development of abnormal diagnosis of diseases through the examination of surgically conditions as well as the structural removed organs and tissues (for biopsy sample). and functional changes that result  Also examines bodily fluids and even the whole bodies in from the disease processes. cases of autopsies.  Defined as the laboratory expert behind the frontline clinical team Medical Perform diagnostic analysis of human Laboratory blood, urine, other body fluids such as Scientist or CSF, peritoneal fluid, pericardial fluid, 2 Branches of Pathology Medical synovial fluid, stool, sputum, and semen. Technologist  From Greek words: auto & opsis AUTO: self 2 Types of Pathology OPSIS: sight  Seeing with one’s own eyes.  Study of basic reaction of cells and tissues to  Aka necropsy or postmortem abnormal stimuli that underlie all diseases. examination General  Common changes in tissues are observed.  Systematic examination of a cadaver for study or for determining the cause o Ex: cancer, aging, inflammation of death.  Study of specific responses of specialized Autopsy  Uses many methodical procedure to organs and tissues to a well-defined stimuli. determine the epidemiology and Systemic  Focuses on “specific changes” in organs. pathogenesis of disease. o Ex: goiter for thyroid gland, pneumonia  Purposes: for lungs, and breast cancer. 1. Family counsel 2. Determine etiology or cause 3. Epidemiological purposes 4. Establishment of genetic causes 5. Pathogenesis of diseases  Examination of cells or tissues from a living organism.  Excised material may be studied to diagnose disease or to confirm findings of normality. Biopsy  Incision can be total or partial.  Partial lesion can remove them in the form of “wedges” or “cylindrical” in shape, or “scraping” off the surface of the membrane of the desired internal organ. Aspects of Diseases Forming the Core of Pathology Etiology Cause Pathogenesis Mechanism of development Morphological Changes Structural alterations Clinical Significance Functional consequences HISTOPATHOLOGIC & CYTOLOGIC TECHNIQUES PRELIM LECTURE | FIRST SEMESTER WEEK 2: INTRODUCTION TO GENERAL PATHOLOGY & HISTOTECHNIQUES 3 Divisions of Pathology  Branch of general pathology which is directed to the diagnosis and monitoring of  The recognition of the disease is based on diseases through the examination of blood, the macroscopic examination (outer large body fluids, secretions, and tissue biopsy appearance) of surgical specimens specimens for chemical, morphological, Gross generated at the time of surgery or microbiological, and immunological Pathology autopsy. abnormalities.  Ex: Brain slices undergoing gross  We identify and interpret changes that pathology. characterized different diseases or disease states inside the cells even the tissues and  The study of changes is in the function, also body fluids. structure, or appearance of the organs or o It will help us to monitor the metabolic tissues, including postmortem status of the patients under medical examinations and the study of biopsy therapy. specimens.  It deciphers specific markers that  Include 3 branches: characterize individual patients for the purposes such as transfusion or  The pathology of disease transplantation. processes that are surgically accessible for  SECTIONS: Surgical diagnosis or treatment. 1. Blood banking 2. Hematology Pathology  Study of gross appearance 3. Clinical chemistry (toxicology) and histology of tissues are 4. Clinical immunology and serology after the removal of the 5. Microbiology (bacteriology, tissue during surgery. parasitology, virology, mycology)  Involves the external and internal examination of a  A division of clinical pathology Anatomic human body after the time involving biochemical analysis Pathology of death. performed on human samples Autopsy Pathology  Study of the gross (blood, fluids, tissues) that are appearance and histology being examined outside of the body of tissues that were (in vitro). removed following death.  Substances that can be assayed:  Branch of general cytology 1. Carbohydrates which deals with the Clinical 2. Sugar microscopic examination of Pathology 3. Enzymes desquamated cells. 4. Lipoproteins o Desquamated Cells 5. Hormones 6. Vitamins  cells that have been 7. Antibodies Exfoliative shed off or removed 8. Lipids cytology from the epithelium 9. Electrolytes  cells that are 10. Metals Clinical Chemistry (toxicology) harvested by rubbing or brushing a lesion  Methods of instrumentation: of epithelial tissue 1. Fluorometry surfaces. 2. HPLC (High Performance Liquid Chromatography) 3. Spectrophotometry 4. Enzyme Kinetic 5. Electrophoresis 6. Flame photometry 7. EIA (Enzyme Immunoassay) 8. Mass Spectrometry 9. Gas Chromatography 10. Ion Selective Electrodes (for electrolytes) TOXICOLOGY  Blood, urine, and other body fluids are analyzed if there are presence of drugs or other substances of abuse.  Done to measure the blood level of therapeutic drugs (response to therapy) o we can assure that the concentrations are already adequate to treat disease.  They are not high or low as to cause toxic side effects  Most instruments are automated, but some are still semiautomated. HISTOPATHOLOGIC & CYTOLOGIC TECHNIQUES PRELIM LECTURE | FIRST SEMESTER WEEK 2: INTRODUCTION TO GENERAL PATHOLOGY & HISTOTECHNIQUES Sections of Clinical Pathology  A division of clinical pathology involved in isolation, culture, and identification of  Involves assessment of the cellular microorganisms (parasites, fungi, elements (RBC, WBC, and platelets) in bacteria, and viruses) in biological blood samples. samples.  Blood cells can be enumerated through  Includes: Parasitology, Mycology, manually counting them or using Bacteriology, and Virology automated counting.  Presumptive identification of microbes  We usually do Microscopic observation can be made by microscopically of stained peripheral blood smear but examining direct amounts of a portion of only limited to assessing the morphology a specimen. Thin smears stained with of typical cells as they may appear in dyes are used depending on the cases of aplastic syndrome also overt organism that will be identified. anemias. Microbiology  Rapid presumptive diagnosis can be done by directly testing a specimen with Hematology a variety of immunologic reagent. HEMATOPATHOLOGIST  Specimens are applied on the surface of  Pathologist that specializes in the field of an agar or culture media for pure culture hematology. to be recovered.  Examines bone marrow and lymph node  Stains: Gram staining biopsies. o Can determine cellular morphology o Important in patients with anemia, and also staining characteristics of leukemia, and lymphomas. bacteria  Bone marrow examination is usually  There are variety of rapid and direct test requested in complicated cases were the that can be performed to provide an diagnosis of hematological disorder early identification. cannot be made by the study of peripheral blood smear alone.  The discipline in which infectious diseases are diagnosed by detecting  Aka immunohematology antibodies in serum and other body  A division of clinical pathology that deals fluids. with collection, storage, compatibility and safety of blood and its various  Immuno logic and serologic techniques components for the purpose of human are used to diagnose an infectious transfusion. disease when that agent is too difficult to recover on a culture.  Other Tasks: Blood 1. Apheresis & Plasmapheresis Banking 2. Investigation of Transfusion (including reactions transfusion 3. Blood collection after donor medicine) screening 4. Immunophenotyping of blood cells 5. Blood typing and screen for antibodies (for compatibility testing) 6. Component preparation and storage 7. Chemical and serologic test (done to exclude the transmission of infectious diseases) Clinical Immunology and Serology o Example: ASO (antistreptolysin O) – blood test to measure antibodies against SLO. Streptolysin O is a substance produced by Group A streptococci. o Some of the illness caused by the bacteria Group A streptococcus are Bacterial Endocarditis (infection on the inner lining of the heart) HISTOTECHNOLOGY  The art and science performed by the histotechnologist to produce a tissue section of good quality that will enable the pathologist to diagnose the presence or absence of a disease. HISTOPATHOLOGIC TECHNIQUE  Involves the different procedures that have been adopted for the preparation of materials and tissue for microscopic examination. HISTOPATHOLOGIC & CYTOLOGIC TECHNIQUES PRELIM LECTURE | FIRST SEMESTER WEEK 2: INTRODUCTION TO GENERAL PATHOLOGY & HISTOTECHNIQUES HISTOLOGY  Branch of biology which studies the microscopic anatomy of  Developed from mesoderm. biological tissues.  Can be found in between other tissues  The microscopic counterpart of gross anatomy. everywhere in our body.  Once examined by the pathologist.  Inside the nervous system, there are 3 outer membranes, called the meninges. o Meninges: envelops brain & spinal cord 4 Basic Types of Animal Tissue  Connective Tissue consists of 3 components:  Main function: for contraction Fibers Can be collagenous,  A soft tissue that composes muscles in elastic, reticular animal bodies and gives rise to muscles’ Ground Watery components ability to contract. Substance Connective  Opposed to other component or tissues Tissue Cells in the muscle such as tendons or perimysium General Specialized Connective Connective Tissue  Formed during embryonic development Tissue through a process known as Muscle a. Loose CT myogenesis. Tissue - Has more ground substance a. Cartilage b. Bone b. Dense CT c. Blood - Has regular and d. Lymph irregular e. Hematopoietic - Has more fibers tissue and less ground substance  Lines the outer surfaces of organs and blood vessels throughout the body, as  Aka Neural tissue well as the inner surfaces of cavities in  Main tissue component of the nervous internal organs. system Nervous system – primarily regulates and controls body functions as ACCORDING TO SHAPE well as activity.  Has cells that has wider Central Peripheral NS than their height. NS Comprise of branching  Flat and scale-like peripheral nerves which is  Usually found in the composed of neurons and Squamous lining of the mouth, neuroglia esophagus, blood Nervous Tissue a. Neurons / Nerve cells: vessels, and alveoli of receives & the lungs. transmits Brain & impulses  Has cells whose height Spinal and width are Cord b. approximately the same. Neuroglia /  Functions: absorption, Glial cells: excretion, secretion. assists in propagation of nerve impulse Squamous-  Can be found in location and provide nourishment Epithelial Cuboidal that is having the same Tissue function.  Can be seen in the collecting ducts of the kidneys, pancreas, and salivary glands.  Cells are taller than they are wide.  Can be further classified into ciliated columnar epithelium or glandular columnar epithelium.  Functions: absorption, columnar excretion, secretion.  Almost has the same function as cuboidal epithelium.  Seen in the respiratory system (example: trachea or bronchi) HISTOPATHOLOGIC & CYTOLOGIC TECHNIQUES PRELIM LECTURE | FIRST SEMESTER WEEK 3: CELLULAR INJURY & CELL DEATH CELLULAR INJURY 7 CATEGORIES OF MOST INJURIOUS STIMULI CELLULAR INJURY  Aka Hypoxia = deficiency of  Results when cells are stressed so severely that oxygen they are no longer able to adapt.  Reduced aerobic oxidative o When cells are exposed to inherently respiration. damaging agents or suffer from intrinsic abnormalities, it could lead to cellular injury.  An extremely important and common cause of cell injury and  Normal cells are confined to a fairly narrow range cell death. of function and also structure by its state of metabolism, differentiation and specialization.  Depending on the severity of the o It is nevertheless able to handle those hypoxic state, the cells can still physiologic demands maintaining a steady adapt, undergo injury, or lead to state called Homeostasis cell death. o Homeostasis – Cells can still adapt o Ex: When an artery is o Normally, when the stress or stimuli causing narrowed, the tissues where the change in the cell is eliminated, the cell it supplies blood may shrink should be able to recover to its original state (atrophy); whereas a more without having suffered any harmful severe or sudden hypoxia consequences. Oxygen may induce cell injury or cell death.  Types of Cellular Injury: Reversible Cell Injury & Deprivation Cell Death (Necrosis or apoptosis) CAUSES Reduced blood ADAPTATION flow in cases of ISCHEMIA  Adaptation of cells to injury are reversible, 1. Inadequate oxygenation of functional, and structural responses to the blood caused by cardio- changes in physiologic state. respiratory failure o Ex: during pregnancy It can also because of 2. Reduced carrying capacity of some pathologic stimuli, during which new blood in cases of anemia but altered steady state are achieved. This 3. Carbon monoxide poisoning 4. Severe blood loss allows the cell to survive and to continue to function. RESULTS IN o It can also because of some pathologic 1. Cell adaptation (depending on stimuli, during which new but altered steady the severity) state are achieved = allows cell to survive 2. Cell death (irreversible) and continue to function. 3. Cell injury (severe) 1. Mechanical trauma 2. Electric shock 3. Radiation Physical 4. Extreme temperatures Agents (burn/deep colds) 5. Sudden changes in the atmospheric pressure (high altitude areas) 1. Virus Infectious 2. Fungi ADAPTIVE RESPONSES Agents 3. Parasite Adaptive response is through an 4. Bacteria Hypertrophy increase on cell size / functional activity  AKA Genetic Disorder or Hyperplasia There’s increase in cell number Aberrations There’s decrease in size &  Have defective genes & genetic Atrophy metabolic activity defects can cause cell injury due Metaplasia Changing cell’s phenotype to deficiency in the protein Genetic Derangements function. CAUSES OF CELLULAR INJURY o Ex: Enzyme defects in Mechanical From an automobile/vehicular inborn errors of metabolism, Trauma accident accumulation of damaged A subtle cellular abnormality, which DNA, misfolded proteins Mutation causes a lack of a vital enzyme that which can all trigger cell impairs normal metabolic function. 1 HISTOPATHOLOGIC & CYTOLOGIC TECHNIQUES PRELIM LECTURE | FIRST SEMESTER WEEK 3: CELLULAR INJURY & CELL DEATH death when they are beyond REVERSIBLE CELL INJURY repair. HALLMARKS 1. Reduced oxidate phosphorylation with Glucose or salt in resultant depletion of energy stores (ATP) hypertonic 2. Cellular swelling caused by changes in ion concentration (can Simple concentration & water influx cause cell injury by chemicals deranging the electrolyte and fluid CHARACTERIZED BY balance in the cell) 1. Blebbing (protrusions) of the plasma Trace membrane Mercuric salts, amounts of 2. Clumping of nuclear chromatin arsenic, and cyanide Chemical poison 3. Detachment of ribosomes from the ER Agents and Industrial 4. General swelling of the cell and its Exposure to carbon Drugs and organelles monoxide or Occupation asbestos Hazards REVERSIBLE CELL INJURY Recreational  Usually occur in early stages or in mild form. Alcohol Drugs  If the damaging stimuli is removed, it will revert Environmental and air on being a normal cell as long as stress can still pollutants, insecticides, and be adapted. herbicides  Checked under the light microscope. Oxygen at high concentrations FEATURES Responsible for CELLULAR SWELLING / HYDROPIC CHANGE / Endogenous VACUOLAR DEGENERATION autoimmune Self-antigen CELLULAR SWELLING diseases Immune  The earliest manifestation of almost all forms of Reactions to injury to cells. many External  The swelling of cells results from influx of water Important that is usually caused by the failure of the Agents like causes of cell adenosine triphosphate (ATP) dependent Immunologic virus, Reactions and tissue injury sodium (Na+) potassium (K+) plasma membrane bacteria, & environmental pump. substances o The failure of the ATP pump occurs  Immune system serves as an because the is already a depletion of ATP essential function in defense due to oxygen deficiency. against the infectious pathogens. o Because of this, the cells are incapable of  Immune reactions may cause maintaining ionic and fluid homeostasis. cellular injury.  Leakage of content will eventually trigger an inflammatory reaction.  Major cause of cell injury ULTRASTRUCTURAL CHANGES OF o Ex: protein-calorie REVERSIBLE CELL INJURY deficiency, deficiency in 1. Blebbing specific vitamins, nutritional Plasma Membrane 2. Blunting excess in cases of obesity, Alterations 3. Loss of microvilli problems in undernutrition or Nutritional Imbalances overnutrition, and 1. Swelling Mitochondrial composition of diet. 2. Appearance of small Changes o High lipid diet leads to amorphous densities elevated serum cholesterol, Dilation of ER, with detachment of polysomes resulting to atherosclerosis Nuclear Alterations (leading risk factor for Disaggregation of Granular & Fibrillar Elements cardiovascular diseases). 2 HISTOPATHOLOGIC & CYTOLOGIC TECHNIQUES PRELIM LECTURE | FIRST SEMESTER WEEK 3: CELLULAR INJURY & CELL DEATH MORPHOLOGIC CHANGES IN REVERSIBLE CELL DEATH CELL INJURY AND NECROSIS CELL DEATH Normal kidney  An event of a biological cell that cease to carry  Cuboidal out its function. tubules (epithelium)  Intact nuclei  A severe and progressive cellular injury that leads to irreversible injury.  Able to see nuclei but there  Cell cannot recover and it dies are irregularities.  From damaged CM and loss of ion homeostasis  Surface blebs  Cellular contents leaks in EC  Inflammatory Reversible  Increased Eosinophilia response (Pinkish in appearance) Lysosomal enzymes enter cytoplasm  Digests cell   Swollen cells Morphological changes (Necrosis)  Loss of Nuclei  Results of natural process of old cells dying, Necrosis  Fragmentation of cells disease, localize injury or death of an organism  Leakage of contents of which the cells a part of.  If cell death is excessive as a result of a progressive injury - it is the most crucial event in the evolution of disease in any tissue or organ. NECROSIS: “Accidental” and unregulated form of cell death TYPES OF CELL DEATH  Programmed cell death.  Is a form of cell death that is generally triggered by the normal and healthy processes in the body FATTY CHANGE FATTY CHANGE Apoptosis  Can also occur as a defense mechanism during healing  Due to: Hypoxic injury and toxic/metabolic injury processes, is almost always  Manifested by the appearance of lipid vacuoles normal and beneficial to an in the cytoplasm. organism. o This results when toxic injury disrupts the metabolic pathway and will later lead to a  Is a cell death that is triggered by rapid accumulation of triglyceride filled external factors or disease, such lipid vacuoles. as trauma or infection.  Often seen in liver because it is the major organ  Necrosis is always abnormal and involved in fat metabolism harmful. Necrosis  Seen in cells involved in and dependent on fat  Considered an “unprogrammed” metabolism (hepatocytes and myocardial cells). (unnatural) cell death process at  Can also occur in the heart, muscle, or kidney. this time. o Historically known as accidental type of cell death APOPTOSIS VS NECROSIS Feature Apoptosis Necrosis Cell Size Reduced (shrinkage) Enlarged Fragmentation into Pyknosis* Nucleus nucleosome-size Karryorrhexis* fragments Karyolysis* Plasma Intact; altered Disrupted Membrane structure Intact; may be Enzymatic Cellular released in apoptotic digestion; may Contents bodies leak out of cell Adjacent No Frequent Inflammation Often physiologic, Invariably Physiologic means of eliminating pathologic or unwanted cells; may (culmination of Pathogenic be pathologic after irreversible cell Role some forms of cell injury) 3 HISTOPATHOLOGIC & CYTOLOGIC TECHNIQUES PRELIM LECTURE | FIRST SEMESTER WEEK 3: CELLULAR INJURY & CELL DEATH injury, especially DNA liberation of enzymes from damage those cells.  Appearance: creamy yellow in CHARACTERISTICS OF NUCLEUS IN NECROSIS color due to presence of dead Reduction in size and condensation of leukocytes (pus) Pyknosis the nucleus + nuclear shrinkage &  Ex: hypoxic death of cells within increased basophilia the CNS that leads to brain Karyorrhexis Fragmentation of chromatin infarction Karyolysis Dissolution of nucleus  Fading of  Refers to chromatin material massive death of the tissue caused by a combination TYPES OF NECROSIS of ischemia and superimposed by  The architecture of the dead bacterial infection. tissue is preserved for a span of Gangrenous  Commonly due to interruption of Necrosis at least some days. blood supply in the lower  Affected tissue has a firm extremities. consistency.  Denatures not only the structural Coagulative Liquefactive proteins but also the enzymes. “Dry gangrene” “Wet gangrene” o Blocks proteolysis of the Arterial occlusion Venous dead cell occlusion o Result: Intensely eosinophilic cells with distinct or reddish nuclei Coagulative can persist for 4 days to Necrosis weeks o Necrotic Cells are broken down by the action of lysosomal enzymes derived from infiltrating leukocytes o Infiltrating Leukocytes it is  Encountered most often in foci of removed by the debris of tuberculosis infection (lungs) dead cells by phagocytosis.  Conversion of destroyed cells into  Seen most often in the heart after granular, friable mass made up of an infarction, as well as in a mixture of coagulated protein kidneys and adrenal glands. and fats. o Example: Ischemia caused  Fragmented or lysed cells by obstruction. enclosed in an inflammatory Caseous border called granuloma. Necrosis  Opposite of CASEOUS: Cheese-like appearance coagulative  Microscopic Examination: necrosis. Structure less, fragmented  Also termed and lysed cells, amorphous as An adaptive response is a coordinated set of actions that an individual exhibits in response granular debris to new or challenging situations. liquefaction  This combination of necrosis. liquefactive and coagulative  Refers to a fairly rapid total necrosis is caused by dead enzymatic dissolution of the cells cells that are not completely Liquefactive Necrosis resulting into complete digested by macrophages destruction of the entire cell.  They leave a granular  Characterized by the digestion of residue that impedes dead cells. circulation  Transformation of the tissues into  Caused by Fungal and viscous liquid mycobacterial infections,  Seen in focal bacteria/fungal such as tuberculosis infections. o Happens since microbes stimulate the accumulation of leukocytes and the 4 HISTOPATHOLOGIC & CYTOLOGIC TECHNIQUES PRELIM LECTURE | FIRST SEMESTER WEEK 3: CELLULAR INJURY & CELL DEATH injury, especially DNA liberation of enzymes from damage those cells.  Appearance: creamy yellow in CHARACTERISTICS OF NUCLEUS IN NECROSIS color due to presence of dead Reduction in size and condensation of leukocytes (pus) Pyknosis the nucleus + nuclear shrinkage &  Ex: hypoxic death of cells within increased basophilia the CNS that leads to brain Karyorrhexis Fragmentation of chromatin infarction Karyolysis Dissolution of nucleus  Fading of  Refers to chromatin material massive death of the tissue caused by a combination TYPES OF NECROSIS of ischemia and superimposed by  The architecture of the dead bacterial infection. tissue is preserved for a span of Gangrenous  Commonly due to interruption of Necrosis at least some days. blood supply in the lower  Affected tissue has a firm extremities. consistency.  Denatures not only the structural Coagulative Liquefactive proteins but also the enzymes. “Dry gangrene” “Wet gangrene” o Blocks proteolysis of the Arterial occlusion Venous dead cell occlusion o Result: Intensely eosinophilic cells with distinct or reddish nuclei Coagulative can persist for 4 days to Necrosis weeks o Necrotic Cells are broken down by the action of lysosomal enzymes derived from infiltrating leukocytes o Infiltrating Leukocytes it is  Encountered most often in foci of removed by the debris of tuberculosis infection (lungs) dead cells by phagocytosis.  Conversion of destroyed cells into  Seen most often in the heart after granular, friable mass made up of an infarction, as well as in a mixture of coagulated protein kidneys and adrenal glands. and fats. o Example: Ischemia caused  Fragmented or lysed cells by obstruction. enclosed in an inflammatory Caseous