Histopathology Lecture Notes PDF

HISTOPATHOLOGY HISTOPATHOLOGY PATHOLOGIC CAUSES o decreased workload—bedrest due to bone fraction Study of; o loss of inn...

HISTOPATHOLOGY HISTOPATHOLOGY PATHOLOGIC CAUSES o decreased workload—bedrest due to bone fraction Study of; o loss of innervations—loss of nerve response impulses ➔ combination of study of histology and pathology o diminished blood supply—ischemia—lack of blood supply ➔discipline utilize all biological sciences that study in tissue—stroke →hypoxia morphology (structures and how disease started) and o inadequate nutrition—carbohydrates food in diet pathology →cachexia—muscle and tissue wasting ▪ Etiology—origin of disease w/ underlying causes →ashimoto thyroid ▪ Pathogenesis—cause of the disease; how the disease o loss of endocrine stimulation—loss of estrogen after progresses menopause; hypothyroidism ▪ Molecular and morphologic changes—different structural o pressure—lead to shrinkage or compression happen on tissues, cells and organs ▪ Clinical manifestations—signs and symptoms of the disease Does atrophy cells are dead? It will just reduce its size but not as well as and how it will progress dead Even there will be different causes why cell undergo atrophy DIVISIONS OF PATHOLOGY there fundamental cellular are identical Low metabolic activity= low CHON synthesis 1. GROSS PATHOLOGY Low nutrients, disease= high CHON degradation ▪ We examine the entire body ▪ Macroscopic—can be seen in naked eye 2. HYPERTROPHY 2. MICROSCOPIC PATHOLOGY ▪ Increase in the size of cells, that result in an increase in ▪ Microscopic manifestation of the disease the size of the affected organ A. ANATOMIC PATHOLOGY—structural and composition changes ➔PHYSIOLOGIC OR PATHOLOGIC o Surgical—getting sample from living person ➔OCCURS CONCURRENTLY WITH HYPERPLASIA o Autopsy –getting sample from deaceased person CAUSED EITHER BY: B. CLINICAL PATHOLOGY 1. Increased functional demand—workout; increased of o Hematology functional demand o Microbiology 2. Specific hormonal stimulation—uterus size after birth; o Clin. Chem. heart due to circulatory overload o Sero/Immuno o Clin. Microscopy—body fluids Is there a present of no cells? Why the organ become big? NO, o Parasitology there is a cell present but it is larger than normal cell because it has additional cell present STAGES OF THE CELLULAR RESPONSE TO STRESS AND INJURIOUS STIMULI It is present to the organ that are not able in replicating and not replicating High metabolic activity= high CHON synthesis 3. HYPERPLASIA ▪ Increase in the number of cells in an organ or tissue in response to a stimulus ▪ An adaptive response in cells capable of replication ▪ May occur with hypertrophy and often in response to the same stimuli 2 TYPES A. PHYSIOLOGIC HYPERPLASIA ✓ Hormonal Hyperplasia o EX. proliferation of glandular epithelium of female breast at puberty and during pregnancy CELLULAR ADAPTATION ✓ Compensatory Hyperplasia ▪ REVERSIBLE changes in the size, number, phenotype, o Occurs when a portion of the tissue is removed metabolic activity, or functions of cells in reponse to or diseased changes in their environment o EX. partial resection of the liver—this is the ▪ Maybe be PHYSIOLOGIC (normal stimulation; basis for living-donor transplant hormonal changes; pregnancy) and PATHOLOGIC B. PATHOLOGIC HYPERPLASIA (cause by injurious stimulus) ✓ Mostly caused by excessive hormonal or growth factor stimulation ADAPTIVE RESPONSE a. ENDOMETRIAL HYPERPLASIA—mens. bleeding; 1. ATROPHY excessive stimulation ▪ Reduction in the size of an organ or tissue due to b. PAPILLOMA—skin warts; abnormal or virus decrease in cell size and number ▪ Shrinkage of cell MAJOR FORMS OF ADAPTION PHYSIOLOGIC CAUSES REMEMBER o Normal development of the body ▪ In all these situations (hormonal or growth factor o Fetus tail in fecal development stimulation), hyperplastic process remains CONTROLLED o Decrease of size of uterus after pregnancy ▪ If hormonal or growth factor stimulation abates, the o Atrophy of brain when aging hyperplasia DISAPPEARS o Brain cells and heart are non replecating ▪ More commonly venous obstruction  HISTOPATHOLOGY ▪ It is this sensitivity to normal regulatory control mechanisms 5. DYSPLASIA that distinguished benign pathologic hyperplasia from ▪ Also referred to as a typical hyperplasia cancer (in which the growth control mechanisms become ▪ Disorder growth and maturation of the cellular dysregulated or ineffective) components of a tissue ▪ EXAMPLE ▪ Cause of injurious stimulus o Pxs with endometrial hyperplasia→ (+) risk of ▪ Early stage of cancerous lesion endometrial CA o Pxs with papillomavirus inxn → (+) risk of cervical CA EPITHELIAL DYSPLASIA o Expansion of immune cells (such as cells of the THE RELATIONSHIP BETWEEN HYPERPLASIA AND ectoderm), with a corresponding decrease in the HYPERTROPHY number and location of mature cells ▪ Although hypertrophy and hyperplasia are two distinct o Dysplasia is often indicative of an early neoplastic processes, frequently both two distinct processes, process frequently both occur together, and they will be o Immature cells are located triggered by the same mechanism o Fertile soil of cancer metaplasia and dysplasia 4. METAPLSIA FOUR MAJOR PATHOLOGICAL MICROSCOPIC CHANGE ▪ Reversible change in which one differentiated cell type ▪ Anisocytosis—presence of cell in unequal size (epithelial or mesenchymal) replaced by another cell ▪ Poikilocytosis—abnormal shape of cell type ▪ Hyperchromatism—excessive pigmentation of cell ▪ Occurs as response to chronic physical or chemical ▪ Presence of mitotic figures—all cell undergo mitotic division irritation ▪ One cell are replaced by another type of cell capable CELL INJUIRY from injurious stimulation → alteration in cell structure or functioning that occur when ▪ The size of cell are not changing cells are: ▪ EX. Changes in the respiratory tract in response of 1. Severely stressed that they are no longer able to adapt inhalation of irritance such as smug or smoke→ciliated 2. Exposed to inherently damaging agents columnar epithelium to non ciliated stratified 3. Suffer from intrinsic abnormalities→inherited squamous epithelium CAUSES OF CELL INJUIRY CAUSES o Changes in environment 1. HYPOXIA o Irritation or inflammation o low level of oxygen in the body o Nutritional o extremely important and one of the causes of cell injury and cell death A. EPITHELIAL METAPLASIA 3 REASONS ✓ Squamous metaplasi—normal lining of esophagus o reduce blood flow lead to ischemia →squamous to columar o patient have cardiovascular failure changes in bronchus o reduce capacity of red blood cells to carry of oxygen uterine endocervix 2. PHYSICAL AGENTS gallbladder, prostate o Mechanical trauma renal pelvis and urinary bladder vitamin A o Temperature (hyperthermia or hypothermia) deficiency o Exposure to radiation lacrimal and salivary glands o Electric shock ✓ Columnar metaplasia—columnar to squamous 3. CHEMICAL AND DRUGS Intestinal metaplasia in healed chronic o Pollutants, hazardous toxins, drug overdose, gastric ulcer alcoholism Barrett’s esophagus—changes from 4. MICROBIOLOGIC AGENTS squamous epithelial to columnar goblet cells o Pathogens B. MESENCHYMAL METAPLASIA 5. IMMUNOLOGIC REACTIONS Osseous metaplasia o Hypersensitivity or allergies Cartilaginous metaplasia 6. GENETIC DEFECTS o Deficiency in the function of proteins—cell not TISSUE NORMAL METAPLASIA STIMULUS function properly AIRWAY Pseudostratified Squamous Cigarette o Damage DNA columnar epithelium smoke o Inborn error of metabolism epithelium 7. NUTRITIONAL IMBALANCES URNINARY Transitional Squamous Bladder stone o Starvation BLADDER epithelium epithelium o Cellular excess Squamous Columnar Gastro- o Malnutrition ESOPHAGUS epithelium epithelium esophageal 8. AGING reflux o Alterations in replication PREDISPOSING FACTORS—susceptible si cell at risk DISEASE 1. AGE ▪ An abnormal condition of an organism or part that ▪ In the process of aging impairs normal physiological functioning, esp. as result of; 2. GENDER 1. Infection—growth of microorganism in the body ▪ Cell injury common in males or females → it will have disease when the cell has injury 3. NUTRITIONAL STATUS 2. Inherent weakness, or 4. GENES 3. Environmental stress 5. ENVIRONMENT ▪ Exposure to chemical and physical agents 6. PRE-EXISTING CONDITIONS 7. IMMUNE-COMPROMISED HISTOPATHOLOGY GENERAL MECHANISMS OF CELL INJUIRY REVERSIBLE CELL INJURY ▪ The cellular response to injurious stimuli depends on: Two features of reversible cell injury (microscopic) ✓ The type of injury 1. CELLULAR SWELLING/ CLOUDY SWELLING—first ✓ Its duration, manifestation of all injury of cells ✓ severity CAUSE: ▪ The consequences of cell injury depend on: o Incapability of maintaining ✓ The type ionic and fluid homeostasis ✓ State and failure of energy- ✓ Adaptability of injured cell dependent ion pumps in the ▪ Cell injury results from: plasma membrane ✓ Different biochemical mechanisms acting on several essential cellular components 2. FATTY CHANGE/ STEATOSIS GENERAL MECHANISMS OF CELL INJURY CAUSE: o Manifested by the appearance Ischemia→ cause from of lipid vacuoles in the hypoxia cytoplasm No oxygen they would o abnormal retention of cell be less oxidative o cell involve in metabolism ( adipose cell, hepatocytes, and phosphorylation and myocardial cells) decrease production of ADDITIONAL FEATURES: ATP a. blebbing of the plasma membrane—cause by intracellular accumulation of ater; asa si sodium naa si Sodium—dominant ion water—causing the cytoskeleton of cell to break—yung inside/ intercellular the water dumadami ang kanyang space walang nag cell memaintain sa shape ni cell result to blebbing b. detachment of ribosomes from ER Potassium—dominant c. clumping of nuclear chromatin ion outside/ IRREVERSIBLE CELL INJURY extracellular the cell 1. increased swelling of cell Pleb—cell structure 2. swelling and disruption of lysosomes 3. presence of large amorphous densities in swollen mitochondria 4. profound nuclear changes: LACTIC ACID—by product of anaerobic glycolysis DECREASE PROTEIN SYNTHESIS—detachment of ribosome’s lead to lower production of protein to utilize in cells IRREVERSIBLE CELL INJURY ROS—reactive oxygen species— counter mechanism of cell to decrease ATP BOTH TWO TYPES OF irreversible cell injury lead to cell Anuclear—no nuclear to be seen due to gramentation death REVERSIBLE VS. IRREVERSIBLE ULTRASTRUCTURAL CHANGES OF REVERSIBLE AND IRREVERSIBLE APOPTOSIS—are CELL INJURY common lead to cell REVERSIBLE INJURY IRREVERSIBLE INJURY death PLASMA Blebbing, loss of Disruption MEMBRANE microvilli MITOCHONDRIA Modest swelling Massive swelling, leakage ENDOPLASMIC Dilation with Extensive disruption and MORPHOLOGIC ALTERATIONS ON CELL INJURY RETICULUM detachment of fragmnetation polysomes FEATURE NECROSIS APOPTOSIS CELL SIZE Enlarged (swelling) Reduced (shrinkage) NUCLEUS Pyknosis, Fragmentation into karyorrhexis, nucleosome-sized karyolysis fragments PLASMA Disrupted Intact MEMBRANE CELLULAR Enzymatic digestion; Intact CONTENTS may leak out of cell ADJACEMENT Frequent No INFLAMMATION  HISTOPATHOLOGY MAY BE CAUSED BY: a. Local bacterial action b. Digestive ferments c. Intracellular enzymes form disintegrated cells d. Chemical and physical agents 2. LIQUEFACTION NECROSIS→ happen in BRAIN o Enzymaticdigestion of the cell by its own hydrolytic lysosomal enzymes o Ischemic brain injury, bacterial infections o Leads to pus formation o Necrosis of tissues rich in liquid usually induces them o to absorb fluid, leading to softening and liquefaction 3. FAT NECROSIS o Happen in our CAVITIES (peritoneal cavity) o Destruction of adipose tissue o Pancreatic lipase splits adipose / neutral fats into FA (fatty acids) and glycerol FEATURE NECROSIS APOPTOSIS o FA combine w/ Ca to form soaps forming white PHYSIOLOGICAL Invariably pathogenic Often physiologic—normal precipitates of CaPO4 and CasCO3 OR PATHOLOGIC ROLE (irreversible) process of cell death o RESULTS: chalky white (pulbos) ACCIDENTAL PROGRAMMED o Digestive tract, liver, and heart Usually affects large Usually affacts scrattered o NODULES—can be seen areas of contiguous individual cells 4. CASEOUS NECROSIS swell o Resemble soft, friable cheese o Cells are converted into a granular, friable, mass of Cells and organelles Cells contact coagulated CHON and fat, with total loss of cell detail swell o Syphilis, Tularemia, LGV, TB Control of intracellular Control of intracellular o Has distinctive inflammatory border environment is lost, environment maintained o Formation of GRANULOMA cells rupture and spill cytoplasm packaged as o Loss of cell detail contents “apoptotic bodies” 5. FIBROID NECROSIS INDUCE DOES NOT INDUCE o Deposition of immune complexes in the walls of INFLAMMATION INFLAMMATION arteries with fibrin that has leaked out of vessels PRINCIPLE TYEPES OF DEATH o Result in a bright pink amorphous appearance of 1. APOPTOSIS fibrinoids o Limits neoplastic o Formation in BLOOD VESSEL WALLS growth—for it to o NEUTROPHILS—seen when it has bacterial infection; lessen and control dominant with fibroid necrosis the cell in our 6. GANGRENOUS NECROSIS body—wait to o Tissue death to ischemia and superimposed bacterial undergo aging our infection cell o Combination of coagulation (INFARCTION) and o Does NOT stimulate liquidation (super in close infarction)necrosis inflammatory 2 TYPES response A. DRY GANGRENE o Goes B. WET GANGRENE fragmentation, and FEATURES DRY GANGRENE WET GANGRENE naturally undergo SITE Commonly limbs More common in bowel due to its normal death cell MECHANISM Arterial occlusion More commonly venous STIMULI THAT ACTIVATE APOPTOSIS: obstruction 1. Cell membrane damage MACROSCOPY Organ dry, shrunken and Part moist, soft, swollen, 2. Mitochondrial damage black rotten and dark 3. DNA damage PUTRFACTION Limited due to very little Marked due to stuffing or 4. Viral infection (bacterial blood supply organ with blood 5. Immune-mediated attack decomposition) ✓ Result to degradation of critical cellular components LINE OF Present at the junction No clear cut line of MECHANISM: activation of enzymes called CASPASES DEMARCATION between healthy and demarcation gangrenous part 2. NECROSIS BACTERIA Bacteria fall to survive Numerous present o Denaturation of intracellular proteins and enzymatic digestion of the lethally injured cell PROGNOSIS Generally better due to Generally poor due to PATTERN TO BASIC MORPHOLOGIC CHANGES (macroscopic little speticemia profound toxemia—can appearance) spread Raynaud’s disease➔ fingers and toes response in cold 1. COAGULATION NECROSIS → lead to HYPOXIA temperature o Denaturation and precipitation of cellular proteins o Occur when arterial supply is cut off, producing anemic ACCORDING TO LOCATION OR EXTENT: or ischemic infarction—solidify becomes firm a. FOCAL—infection is in small scattered area o INFARCT—area of the tissue w/ coagulation necrosis b. MASSIVE—infection is in large area of the body o Happen in HEART infarction HISTOPATHOLOGY OTHER FORMS OF CELLULAR RESPONSES AND INFLAMMATION E. PIGMENTS AUTOPHAGY CAUSES ▪ Cell eating a. EXOGENOUS PIGMENTS—external ▪ Starved cell cannibalizes itself and recycles the digested ❑ Carbon (coal dust) most common contents ❑ Anthracosis ▪ Regulated by a define set of autophagy genes (Atgs)→ ❑ Pneumoconiosis genes ❑ Tattooing ❑ Silicosis b. ENDOGINOUS PIGMENTS ❑ Lipofuscin—known as lipochrome where pigment ; re-radical injury and lipid peroxidation ❑ Melanin—give color pigment ❑ Hemosiderin—HEMOSIDEROSIS—accumulate high hemosiderin when it has hemolysis PATHOLOGIC CALCIFICATION ▪ Abnormal tissue deposition of calcium salts ▪ Smaller amounts of Fe, Mg, and other minerals salts DYSREGULATIONOCCURS IN: ▪ DYSTROPHIC (locally in dying tissues; serum level calcium 1. Cancers is normal and no disruption of calcium metabolism ) OR 2. Inflammatory bowel diseases METASTATIC (in abnormal tissue, high level of serum 3. Neurodegenerative disorders calcium, and disturbance in calcium metabolism) 4. Infections CELLULAR AGING INTRACELLULAR ACCULMULATIONS ▪ Result a progressive decline in cellular function and ▪ Metabolic arrangement of cell of abnormal amount of cell viability caused by: that may be harmless or associated with various decrease 1. Genetic abnormalities of injury 2. Accumulation of cellular and molecular damage ▪ Disposition of different substances intracellular ▪ Occur in lysosome or nucleus of the cell MECHANISM OF INTRACELLULAR ACCUMULATION 1. ABNORMAL METABOLISM—normal endogenous substance that produce at the normal and increase rate but the rate of metabolism is inaccurate/inadequate of metabolites 2. ALTERATIONS IN PROTEIN TRANSPORT AND FOLDING — abnormal endogenous substance that might produce from imitating genes result to accumulation of defective proteins may abnormalities in transport or folding 3. DEFICIENCY OF CRITICAK ENZYMES—chemical reaction take place it will not possible without enzymes present; lack of enzymes to metabolize it only accumulate inside the cell SUMMARY OF CELLULAR RESPONSES TO INJURY 4. INABILITY TO DEGRADE OR TRANSPORT THE SUBTANCE— INJURIOUS STIMULI CELLULAR RESPONSE accumulate inside the cell due to lack of enzyme Altered physiological stimuli; Cellular adaptation machinery nonlethal injurious stimuli PROTEIN—if wala ang carrier hindi siya ma eliminate ❑ Increased demand, ❑ Hyperplasia instead it only accumulate inside the cell increased stimulation ❑ Hypertrophy ❑ Decreased nutrients, ❑ Atrophy A. LIPIDS decresed stimulation ❑ metaplasia ▪ CAUSES OF STEATOSIS—toxins, protein malnutrition, ❑ Chronic irritation diabetes mellitus, obesity, and anoxia REDUCED OXYGEN SUPPLY; CELL INJURY ▪ CAUSES OF CHOLESTEROL ACCUMULATION— CHEMICAL INJURY; MICROBIAL atherosclerosis, Niemann-Pick disease (type C) INFECTION B. PROTEINS ❑ acute and transient ❑ Acute reversible injury ▪ CAUSES ❑ Renal diseases associated with protein loss ❑ progressive and severe ❑ Irreversible injury→ cell ❑ Defective intracellular transport and secretion of (including DNA damage) death critical proteins ❑ Necrosis/ apoptosis ❑ Accumulation of cytoskeleton proteins Metabolic alteration, Genetic or Intracellular Accumulations; ❑ Aggregation of abnormal proteins Acquired; Chronic Injury Calcification C. HYALINE→ transparent ❑ Cumulative Sub-lethal Injury ❑ Cellular Aging ▪ CAUSES: Over Long Life Span ❑ Reabsorption droplets CYTOPATHOLOGIC CHANGES IN DISEASES ❑ Russell bodies—larger eonophilic inclusion ❑ Alcoholic hyaline INFLAMMATION ❑ Hyalinization of walls of renal arterioles in long ▪ a universal to tissue damage by a wide range of harmful standing HTN and DM stimuli including mechanical trauma, tissue necrosis and ❑ Abundant in hyaline tissues infection D. GLYCOGEN ▪ natural reaction of the body which part of immune CAUSES: response in the body ❑ Abnormality in either glucose or glycogen FUNCTIONS metabolism ❑ contain and isolate the injury ❑ Diabetes Mellitus ❑ destroy microorganisms/ toxins ❑ Glycogen storage diseases—appear clear vacuoles ❑ prepare tissue for healing and repair in cytoplasm  HISTOPATHOLOGY EVENTS IN INFLAMMATION CYTOPATHOLOGIC CHANGES IN DISEASES 1. VASODILATION STEPS OF INFLAMMATORY RESPONSE ▪ Change in size of caliber of blood vessels 1. Histamine and postagladins released Involve 2. Capillaries dilate clotting begins ✓ Histamine—increased blood vessel permeability 3. Chemotactic factors attract phagocytic cells release by mast cells 4. Phagocytes consume pathogens and cell debris ✓ PROSTAGLANDINS ✓ NITRIC ACID 2. ENDOTHELIAL ACTIVATION ▪ Increased vascular permeability ▪ Edema formation—swelling; water was permeable in our blood vessles Substance Involved ✓ Histamine ✓ Anaphylokosine ✓ Kinins ✓ Leukotins ✓ PaF 3. NEUTROPHIL ACTIVATION ▪ WBCs enter site of injury CARDINAL SIGNS OF INFLAMMATION ▪ Kill organisms, mo debris CHEMICAL MEDIATORS OF INFLAMMATION CARDINAL SIGN PHYSIOLOGICAL RATIONALE 1. HISTAMINE—increased vessel permeability; mast cells RUBOR (REDNESS) Increased blood flow 2. BRADYKININ TUMOR Exudtion of fluid ❑ Increased vessel permeability (SWELLING) ❑ Causes pain; from plasma protein CALOR (HEAT) Increased blood flow, exudation of fluid, released 3. COMPLEMENT SYSTEM of inflammatory mediator ❑ Group of plasma proteins for immune defense DOLOR (PAIN) Stretching of pain receptors, and nerves by ❑ Min driver is our antibodies and phagocytic cells inflammatory exudates, chemical mediators and attack plasma membrane of phagocytes FUNCTIO LAESA Pain, disruption of tissue structure, fibroplasias 4. ARACHIDONIC ACID DERIVATIVES (LOSS OF and metaplasia ❑ From phospholipids in cell wall FUNCTION) SCUB→no function in our tissue ❑ Leukotrienes, prostaglandins, etc VASOLIDATION Prostaglandin I. ACCORDING TO DURATION Nitric Acid A. ACCUTE/ EXUDATIVE Histamine o Sudden onset INCREASED VASCULAR Vasoactive amines o With vascular and exudative changes; PMNs PERMIABILITY C3a and C5a (through liberating amines) (polynuclear cells—multiple nuclei present; Bradykinin neutrophils) Leukotrienes C4, D4, E4 B. CHRONIC PAF o Involves persistence of injurious agent Substance P o Proliferative response; mononuclear cells— CHEMOTAXIS, C5a lymphocytes—single nucleus LEUKOCYTE Leukotriene B4 INFLAMMATION—accumulation of fluid in the cavity called RECRUITMENT AND Chemokines EFFUSION—accumulation in the site of the infection or in the ACTIVATION IL-1, TNF cavities in the body Bacterial products CYFUSION (two types—transxudase—ultra filtric plasma with FEVER IL-1, TNF little protein and few cell—apperaance grossly clear and Prostaglandins exudates—but rich in protein and more cells—appearance PAIN Prostaglandins grossly cloudy) Bradykinin TISSUE DAMAGE Neutrophil and macrophage lysosomal II. ACCORDING TO CHARACTER OF EXUDATE: enzymes ❑ SEROUS—serosa Oxygen metabolites →tranxudate clear Nitric oxide appearnce INFLAMMATION: CELLULAR COMPONENTS ❑ FIBRINOUS— NEUTROPHIL ❑ Phagocyte, first to enter site of injury, serofibrinous short lived →exudat—compose of ❑ Produce chemicals to attract other cells fibrin strand → ❑ Acute inflammation increase amount of EOSINOPHIL ❑ Longer lived, phagocyte fibrinogen ❑ Allergic rxns, parasitic infections and ❑ CATARRHAL— chronic inflammation exudate—present in BASOPHILS/ MAST ❑ Release histamine surface of epithelium— CELLS increase amount of MACROPHAGES ❑ Major phagocytes, enter site 3-4 days inflammation— after injury increase amount of ❑ Chronic inflammation fluid LYMPHOCYTES/PLAS ❑ Immune fxn in chronic inflammation ❑ HEMORRHAGIC— MA CELLS serosanguinous→infusi on of RBC ❑ Suppurative/purulent— Pus present—more PMNs present HISTOPATHOLOGY III. ACCORDING TO LOCATION DEVELOPMENTAL DEFECTS: ▪ Localize—located in very specific area of the body—do A. APLASIA not spread ❑ From the greek word “aplases”—not molding ▪ Generalize or systemic→ entire area was infected ❑ Defective development or congenital absence of an organ EFFECTS OF ACUTE INFLAMMATION or tissue with primordium ❑ Incomplete tissue development lead to incomplete BENEFICIAL EFFECTS formation of an organ o Dilution of toxins ❑ ACC—aplasia cutis congenital—skin defect characterized o Entry of antibodies by a focal or extensive absence of the epidermis, dermis, o Drug transport and subcutaneous tissue; affects in the scalp o Delivery of nutrients and oxygen ❑ APLASTIC ANEMIA—inability of stem cell to generate o Fibrin formation mature blood cell→pancytopenia(decreased WBC and RBC o Stimulation of immune response and platelets) HARMFUL EFFECTS ▪ Digestion of harmful tissues B. HYPOPLASIA ▪ Swelling ❑ Failure of an organ/ tissue to reach its normal size (less ▪ Inappropriate inflammatory response severe than aplasia) SEQUELAE OF ACUTE INFLAMMATION ❑ Dili na mature ang growth sa organ ❑ EXAMPLE: Congenital Tibia Hypoplasia—CCH Possible outcomes of acute inflammation are: ✓ Resolution C. ATRESIA ✓ Organization and repair ❑ Absence or abnormal narrowing of an opening or passage ✓ Suppuration (abscess formation) in the body ✓ Chronic inflammation ❑ MICROTIA—congenital deformity of the outer ear where Depend on the ear does not fully develop during first trimester of o Type of tissue involved pregnancy o Amount of tissue destruction o Nature if injurious agent D. AGENESIA POSSIBLE OUTCOMES ❑ Complete failure of an organ/ tissue to develop with no 1. RESOLUTION associated primordium Occurs when: ❑ PENILE AGENESIS—urogenital tract malformation with ❑ Connective tissue, framework is complete congenital absence of the phallus; non ❑ Involved tissue has the capacity to replace any production of reproduction hormone→induce hormone testosterone to have normal structure of reproductive specialized cells that have been lost system 2. HEALING BY ORGANIZATION ❑ MAYER-ROKITANSKY-KUSTER-HAUSER-SYNDROME Occurs when: (MRKH) or know as Mullerian Agenesis→not well ❑ Substantial damage to the connective tissue develop or missing vagina or missing uterus framework; and/ or ❑ Tissue lacks the ability to regenerate specialized cells 2. DEGENERATIVE CHANGES—abnormal type of cells that includes its processes that will cause the tissue deturiation and lead to loss of function of the cells from traumatic injury, aging, HEALING BY FIBROSIS/ORGANIZATION well and care Removal of dead tissues and acute inflammatory exudates from damage area (macrophage) A. DYSPLASIA ❑ Malformation Filling of defect by ingrowth of a specialized vascular connective ❑ ↑ immature cells ↓ mature cells tissue called GRANULATION TISSUES (ORGANISATION) ❑ Delayed cell maturation and differentiation Gradual production of collagen by granulation tissue to form a ❑ Often indicative of an early neoplastice process— fibrous (collagenous) scar constituting the repair process PRECANCER 4 MAJOR PATHOLOGIC MICROSCOPIC CHANGES Re-establishment of structural integrity 1. Aniscytosis 2. Poikilocytosis 3. ABSCESS FORMATION 3. Hyperchromatism Occurs when: 4. Presence of mitotic figures ❑ When the acute inflammatory rxn fails to destroy/remove the cause of tissue damage B. ANAPLASIA Mechanism ❑ Development of cell towards more primitive cell types Progression of acute inflammation ❑ ADULT CELL→ embryonic cell type (baliktad) ❑ Loss of differentiation Liquefaction of tissue to form pus ❑ Occur in malignant tumor At the periphery of this area, a chronic inflammatory component C. NEOPLASIA surrounds the area ❑ No growth Fibrous tissue is laid down ❑ CAUSES: carcinogens ❑ DNA alterations Walling of the suppuration (covers the pus) (focus only in the ❑ Transformation of prto-ocogenes (help to regular cell infection of inflammation in isolation) growth differentiation) into oncogenes (tumor inducing 4. CHRONIC INFLAMMATION genes) / loss of tumor suppressor gene infection ❑ May result following acute inflammation ❑ ANTI-ONCOGENE—tumor suppressor gene that counter act ❑ Persistence of injurious agent the oncogene that protect a cell para dii mag grow ABRNORMAL IN CELL GROWTH AND SOMATIC DEATH ❑ Uncontrolled growth →disturbances of the cell which might affect the growth of cells that GENERAL CHARACTERISTICS OF TUMOR CELLS might range from complete absences of tissue to toally unregulated ▪ Usually resemble and imitate normal cell wall enough growth of cells ▪ Capable of secreting substance ▪ PARASITIC NATURE; competes with cells for metabolic needs 1. RETROGESSIVE CHANGES—includes all cell abnormalities which ▪ AUTONOMY; non-responsive to normal growth controls characterize by environmental defect ▪ Steadily ↑ size regardless of environment and host’s health → abnormal growth and tissue development HISTOPATHOLOGY PARTS OF NEOPLSTIC TISSUE A. PRIMARY CHANGES 1. CIRCULATORY FAILURE A. PARENCHYMA (MEDULLARY) ❑ Cardiac function ceases ▪ Active elements of tumor ❑ Absences of pulse and heart beat ▪ Made up of cells which proliferate excessively 2. RESPIRATORY FILURE ▪ Stimulation of abundant collagenous Stroma ❑ Following cardiac failure →DYSMOPLASIA ❑ Lead to death (absences of O2 and ↑ CO2) ❑ Loss of oxidative process B. STROMA (SCHIRRHOUS) 3. NERVOUS FAILURE (CNS FAILURE) ▪ Connective tissue framework with lymphatic and vascular ❑ Loss of reflexes channels CESSATION RESPIRATION→ no breathing after 3 minutes ▪ Supply nutrients CESSATION CIRCULATION→ no heartbeat after 5 minutes CLASSIFICATION OF TUMORS B. SECONDARY CHANGES 1. ALGOR MORTIS ❑ Cooling of the body BASED ON CAPACITY TO PRODUCE DEATH ❑ Body temperature decreases at 7 F/ hr 1. BENIGN TUMORS ACCELERATED BY: ❑ Do not usually produce death ✓ Cold weather ❑ More common in younger age ✓ Severe hemorrhage ❑ Group grows slowly by expansion not fixed ✓ After long wasting diseases ❑ METASTASIS (-) ✓ In lean, malnourished, dehydrated individuaks ❑ NO CACHEXIA—shrinkage of muscles or weight SLOWED DOWN BY: ❑ Capsulated, does not affect status ✓ Consumption of drugs, extreme physical activity, fever 2. MALIGNANT TUMORS THE GLAISTER EQUATION—estimates the hours elapsed since death ❑ Cause death as linear function of the rectal temperature: ❑ Found in older age group 2. RIGOR MORTIS ❑ Grows rapidly by filtration or by expansion ▪ Rigidity or stiffening of muscles ❑ Fixued to tissues ▪ 6-12 hours after death and persists for 3-4 days ❑ Ultimate death by cachexia ▪ Head and neck then towards lower extremities ❑ METASTASIS (+)—irregularly growth ▪ Muscular activity at time of death affects body position BENIGH TUMORS MALIGNANT TUMORS ▪ MECHANISM: ACTIN, MYOSIN FIBERS BIND TOGETHER ✓ Normal cells ✓ Undifferentiated cells FORM JELLY LIKE STRUCTURES (differentiated) ✓ More mitotic figures 3. LIVOR MORTIS (POSTMORTEM LIVIDITY) / ✓ Less mitotic figures ✓ Hyperchromic SUGGILLATION ✓ Not hyperchromic ✓ More tendency to ✓ Less tendency to hemorrhage a nd necrosis ▪ Purplish discoloration of skin due to stasis hemorrhage and necrosis ▪ Discoloration in contrasts to ecchymosis ▪ Absent when blood clots are found in the insterstitial BASED ON HISTOLOGIC CHARACTERISTICS tissues 4. POSTMORTEM CLOTTING A. CARCINOMA ▪ Occurs slowly, immediately after death ▪ Malignant tumors of epithelial origin ▪ Seen fter death and autopsy ▪ Less tendency to produce supporting tissue or stroma ▪ Settling and separation of RBCs from fluid phase of B. SARCOMA blood; RUBBERY with RBCs➔ “CURRENT JELLY” ▪ Malignant tumors of connective tissue origin CLOTS ▪ (+) intracellular tissue framework 5. DESSICATION ▪ Drying and wrinkling of fluid –filled organs GRADING OF TUMORS ▪ Cornea and anterior chamber of the eye ▪ Attempst to establish an estimate of the level of malignancy 6. PUTREFACTION ▪ Based on cytologic differentiation of tumor cells and number of ▪ Loss of rigor mortis due to autolysis of cells mitoses w/in the tumor ▪ Foul-smelling odor due to bacterial invasion ▪ Guide for treatment and prognosis ASSOCIATED WITH THE FOLLOWING CHANGES: ▪ Well-differentiated tumors as a rule are less malignant that ❑ Greenish blue discoloration (Fe sulfide formation) undifferentiated cells ❑ Refraction of cornea ❑ Loss of Rigor Mortis BASED ON: ❑ Peeling of skin and swelling of face ❑ Degree of differentiation of the tumor cells ❑ Followed by ADIPOCERE FORMATION—wax like ❑ Degree of pleomorphism organic substance ❑ Mitotic index ❑ Degree of anaplasia or undifferentiation A. Clostritidium perfringes (welschii) ❑ An estimate of the growth B. Staphylococcus sp., ❑ Degree of invasion C. Streptococcus sp., BRODER’S CLASSIFICATION D. Bacillus., GRADE DIFFERENTIATED UNDIFFERENTIATED 7. AUTOLYSIS CELLS CELLS ▪ Self digestion of cells 1 100%-75% 25%-0% ▪ Eventually undergone by all tissues 2 75%-50% 50%-25% ▪ Putrefaction bacterial from GIT diffuse bacteria from 3 50%-25% 75%-50% GIT diffuse into surrounding tissues and enhance 4 25%-0% 100%-75% destruction of cells SOMATIC DEATH ▪ Progressive dessication, putrifaction and autolysis →total inversible cessation of celebral function, spontaneous will eventually produce total digestion of soft tissues function of respiratory system and circulatory system HISTOPATHOLOGY INSTRUMENTATION IN HISTOTECHNOLOGY—LABORATORY 5. CRYOSTAT OR COLD MICROTOME→ for cutting frozen sections MAJOR EQUIPMENT FOR ANY SURGICAL LABORATORY 6. ULTRATHIN MICROTOME→for cutting sections for Electron 1. Microscope micrscopy 2. Microtome AUTOTECHNICON 3. Cryostat o Autotechnicians make simple processing quick and painless 4. Autotechnicon o The various solutions used for processing tissue are placed into 5. Automated coverslip ten separate nylon breakers 6. Automated H & E stainer o These beakers are placed on the circular deck of the NOTE: instrument. There are also two paraffin baths that are mounted o First and most important step in the operation of any piece of on the deck equipment is to “READ THE MANUAL” that accompanies the o Each paraffin bath contains as internal heater that is equipment thermostatically controlled o NEVER attempt to set up a major piece of equipment without KEY FEATURES approval from the manufacturer 1. Easy to use o Always learn the basics of any machine’s operation before 2. Complete dehydration series to prepare for paraffin using embedding o Each piece of equipment has its own checklist which is 3. Automatic operation—minimal human interaction administered by supervisor or senior staff AUTOMATED COVERSLIP o PURPOSE OF CHECKLIST ▪ Automated coverslip offers the most advanced technology ✓ Prescribe the appropriate use of equipment available to mke coverslipping as effortless as possible ✓ Ensures that the laboratory equipment use safely ▪ Slides are immersed in solvent to minimize drying and ensure and effectively an instant, secure bond when tissue-Tek coverslipping fil is A. File should contain information it involve; applied o Name ▪ After coverslipping, slides are held in a carousel with a full o Manufacturer capacity of 240 slides o Model number AUTOMATED STAINER o Serial number ▪ Automated slide staining B. Record of preventive maintenance perform as prescribe by the ▪ Able to stain up to 100 slides per hour manufacture ▪ Eliminates labor-intensive manual staining C. Record of service calls and repairs perform ▪ Protocols can be saved and shared across multiple devices D. Copy of operative manual via USB drive for complete slide staining standardization MICROSCOPE ARCHIVING AND STORAGE HISTOPATH REPORTS ▪ The microscope is one piece of equipment that used by both 1. Surgical pathology the pathologists and the histo technologists 2. Cytopathology report ▪ The microscope enlarges images and allows the visualization of 3. Autopsy report the morphologic cellular details that are too small to be seen by 3 copies were made/released per test the naked eye ✓ 1 for patient (original) ▪ With the aid of lenses, the unstained section allows the ✓ 1 for safe keeping (file) majority of light pass though, but without adequate distinction ✓ 1 for physician between various tissue structures SIGNATORIES ▪ Stains and dyes are used to give contrasts to the tissue by 1. Request Form→ (from patient’s physician) creating light absorption of varying degrees, uniquely taken up 2. Result Forms → (Pathologist) by each tissue element, and seen microscopically as colors. SPECIMEN HANDLING TO BE USEFUL, A MICROSCOPE MUST ACCOMPLISH THREE 1. Fix First→ using fixatives available THINGS 2. Label 1. It must magnify the object STORAGE OF SPECIMEN (TISSUE BLOCKS AND SLIDES) 2. It must resolve the details of the object o SPECIMEN—1 month to 1 year 3. It must make these details visible o TISSUE BLOCKS—3 months to 10 years TYPES OF MICROSCOPE o SLIDES—indefinite 1. Compound microscope ARCHIVING 2. Bright field microscope ✓ All results are kept on file and some are LIS archived already 3. Dark field microscope ✓ Request for a second copy is a must before claiming the result 4. Phase contrast microscope→ confirmatory and detection of erythromal lupus si patients RECORDS 5. Fluorescence microscope 6. Electron microscope RECORDS/SPECIMEN TYPE RETENTION—years or months wherein they must be kept or destroyed HISTOTECHNOLOGITS→ more equip and trained in using microscope Request 2 years PATHOLOGIST→ examined id the tissue have a disease Quality control 2 years MICROTOME Instrument Maintenance 2 years Blood bank/ Recipient Records Indefinitely ▪ MICROTOME→ (from the Greek micros, meaning “small”, and Blood Bank Employee Signatures/ 10 years Initials temnein, meaning “to cut” ▪ MICROTOMY→ is process by which processed tissue, most Blood Bank Quality Control 5 years commonly a paraffin embedded tissue, is trimmed and cut into Clinical Pathology Laboratory 2 years uniformly thin slices or “sections” Results ▪ MICROTOMES→ are used in microscopy, allowing the Autopsy Forensic Reports Indefinitely preparation of sample for observation under transmitted light Surgical (and Bone Marrow Reports) 10 years or electron radiation Cytogenetics Reports 20 years KINDS OF MICROTOME Serum/ body fluids 48 hours 1. ROCKING MICROTOME→ to cut serial sections of large blocks Blood smears—routine 7 days of paraffin (composed of paraffin wax consist of hydrocarbon) Pathology/ Bone marrow slides 10 years embedded tissues Pathology blocks 10 years 2. ROTARY MICROTOME→common use in laboratory; for cutting Microbiology smears 7 days paraffin embedded sections Blood bank donor/ Recipient 7 days post-transfusion 3. SLIDDING MICROTOME→for cutting celloidin (semi-solid of specimens pyroxylene→soluble, ether and alcohol) embedded sections Cytogenetics slides 3 years 4. FREEZING MICROTOME→for cutting unembedded frozen Cytogenetics diagnostic images 20 years sections HISTOPATHOLOGY MIDTERM TOPICS HISTOTECHNOLOGIST/ HISTOTECHNICIAN ― Quality in Histopth laboratory ― Ensures formalin and other chemicals are fresh and in good ― Pre-analytical factors working qulity ― Specimen collection and handling ― Regularly filters HEMATOXYLIN and other reagents ― Routine tissue processing techniques ― Maintains equipment in high quality condition > Fixation → Preventive maintenance (no overlading) > Decalcification ― Works systematically to minimized errors > Dehydration ― Analyzes problems and corrects them > Clearing ― Provides slides that are properly labeled, processed stained, > Infiltration mounted and sequenced > Embedding SERVICES > Trimming ― Tissue processing > Sectioning ― Cytology > Staining ― Frozen biopsied sections—specimens needed to examined > Mounting immediately > Ringing ― Special staining (freeze drying → Viable for microscopic examination ― Immunostaining → Responsible to examine microscopically the tissue DOCUMENTS May be stored numerically, alphabetically or chronologically QUALITY IN THE HISTOPATHOLOGY LABORATORY TYPES OF DOCUMENTS QUALITY ASSURANCE 1. REQUEST FORM It means of: ― Patients name, age, sex, dob, room/bed/OPD ― Getting the right test ― Hospital/lab accession number ― At the right time ― Specimen type/source; clinical impression ― On the right specimen ― Pertinent history; operative findings ― From the right patient ― Test requested; procedure performed ― With the right diagnosis ― Date and time of request, collection and transportation ― At the right price ― Requesting physician > THINK QUALITY: Our Aim is ZERO defects 2. PATIENT’S REPORT (conveys the diagnosis) > It includes ― Same patients as requested form; plus → Availability of reagents/ supplies → DIAGNOSIS: gross/microscopic findings → Preventive maintenance and monitoring of → NAME OF PTHOLOGISTS equipments 3. TELEPHONE REPORTS → Evaluation of the quality of services—important as ― Preliminary diagnosis the basis if we aim and achieve our desired result ― FINAL DIAGNOSIS will be sent out in your report form QUALITY MANAGEMENT SYSTEMS 4. PRELIMINARY REPORTS ― A set of coordinated activities to regulate a laboratory in ― Status of results 48-72 hours from receipt of specimen order to continually improve its performance 5. FINAL REPORTS ― Continuously improving ― Conveys result after test is completed FACTORS: 6. CORRECTED REPORTS > Skilled histotechnologists/ histotechnician ― Inconsistent report form needed to be clarify > Proper specimen collection and processing 7. INCIDENT REPORTS > Efficient processing of results and documentation ― Documents problems related to personnel > High quality of reagents and equipment performance or other complaints > Preventive maintenance of equipment—regularly ― Involve are the staff of the hospital calibrated * Blocks or tissue slides can be stored in the lab for at least 10 > Continuous professional education of staff years HUMAN RESOURCE MANAGEMENT PRE-ANALYTICAL FACTORS IN HISTOPATHOLOGY LABORATORY PATHOLOGIST: head of the laboratory PRE-ANALYTICAL FACTORS ASSOCIATE PATHOLOGIST: supports the head > Events resulting in biochemical changes that lead to poor tissue quality FUNCTIONS: > Before delivering/ arriving the specimen in the laboratory ― Pinpoint problematic situations and find solutions or preventive measures 1. WARM ISCHEMIA ― Only accept sections that are properly processed, ― Initial anoxic insult in tissues when blood supply is cut stained, labeled and sequenced off ― Monitor staff performance ― Duration depends on circumstances of surgery ― Inspect patients for frozen sections, formalin fixation, ― Beyond the control of the histopath lab aliquot tissue for flow cytometry or fixation in ― Interruption of blood supply in the organ—remove glutaraldehyde for EM from the body—before butnga ug hypotraumic ― open or section hollow tissues to allow fixation of solution fixative and forestall autolysis. 2. COLD ISCHEMIA * Annually review ― Hypoxic event seen in tissues preserved in * Conducting gross examination of tissue depending un hypothermic state until it is processed diameter and size ― Preserved in flush hyperthermic solution until * ASSITANT: medical Technologist transplant > If the associate pathologist is a grosser, he/ she shall COLD ISCHEMIA TIMES ― Identify and match specimen with request HEART 3-4 ― Describe specimen in detail (drawing/photo) LUNGS 3-5 ― Take 3mm sections and label properly LIVER Accessory digits siya > Bunions and hammer toes—common deformity ― EVISCERATION > Extraocular muscle from corrective surgery—eyes → Removal of thoracic and abdominal organ blocks prior > Inguinal hernia scas in adults to gross examination > Nasal bone and cartilage from rhinoplasty—ilong → Removing or organ in sequence the organs are weigh > Prosthetic breast implants and describe > Prosthetic heart valves without attached tissue → Weigh after and describe > Tonsils from children > Umbilical hernia sacs in children GROSS EXAMINATION > Varicose veins ― Consists of describing the specimen and placing all parts of SPECIMENS THAT MAY BE EXCLUDED FROM MANDATORY it into a plastic cassette as it is being processed to a SUBMISSION TO HISTOPATH LAB paraffin block ― Bone donated to the bank bank ― ROLE: Pathologist describe the weight, size, diameter of ― Bone segments removed as part of reconstructive the tissue sample orthopedic procedures ― ROLE: Histotechnician is to take note the description of the ― Fat removed by liposunction pathologist ― Foreskin from circumcision ― GROSSING TABLE: consists of scissor, scapel, ruler, pencil ― Bullets or other medico-legal evidence fiven directly to law DISSECTING PROPS enforcement personnel ― Cutting board ― IUDs without attached soft tissue ― Forceps ― Middle ear ossicles ― Ruler ― Saphenus vein segments harvested from coronary artery ― Scapel/ Knives/ saw bypass ― Inks/ dyes ― Skin or other normal tissie removed during cosmetic or ― Cassettes/ lids reconstructive procedure (not lesion or the patients does ― Bipsy pads/ tissue not have a history of malignancy) ― Filter bags ― Normal toenails and fingernails that are incidentally ― Weighing scales removed ― Wash area SPECIMEN DISSECTION SPECIMEN CATEGORIES 1. BREADLOAFING TECHNIQUE > Incomplete parallel cuts minimum of 2 cm apart A Specimens only requiring transfer from container to tissue perform on large specimens to ensure that cassette pathologic areas or tumoral areas are identified B Specimens are requiring transfer but with standard > Lahat ng parts are able to identified sampling, counting, weighing or slicing > Common method for surgical examination in ― Moderate assessment histopath C Simple dissection required with sampling needing IMPORTANT THINGS TO NOTE dissection required with sampling needing a low level of a. All hollow structures must be opned diagnostic assessment and/ or preparation b. Specimen edges should be squared ― Simple assessment c. Specimens must be fit easily into the cassette—tissue D Dissection and sampling required needing a moderate size not be more than 3 cm in thickness in order for the level and assessment fixative penetrate in the tissue E Specimens requiring complex dissection and sampling d. embed paper tags inside the cassette—graphite of the methods pencil cannot be erase when staining ― Complicated assessment e. Wrap minute and multiple tissue fragments—wrap in ― Cancerous dissection toter paper CATEGORY A SPECIMENS f. Indicate the number of sections of blocks in the specimen A. g. If the specimen is entirely submitted for processing, the ― Endometrium specimen containers are saved until the case is signed ― Breast core biopsies WHAT DOES THE PATHOLOGIST NEED TO KNOW? ― Colonic series Provide good descriptions B. ― Say what you see! ― Small lipoma > Shape ― Small skin bipsy > Color ― Cervical LLETZ (large loop excision of transformation > Weight—runded to the nearesr 0.1 gram zone)―part of the cervix na nay change of the type of cell > Dimensions—rounded to the nearest 1.o cm present > Consistency C. > Distances from margin (s) ― Prepuce > Orientation markers ― Gallbladder > Cut surface appearance ― Haemorrhoids SPECIMEN SAMPLING ― Appendix ― Literally, taking a sample of the tissue D. ― Representative, of the tissue being submitted ― Pigmented skin lesions ― Generally, fewer blocks required if the tissue looks uniform ― Large intestine (Crohn’s) throughout (for benign cases—carcinoma ) ― Skin with markers ― Salivary gland tumour HISTOPATHOLOGY SAMPLING RULES EXAMPLE: prostate chips (19 grams) - SINGLE SUTURE—lateral margins > Sampled in 8 cassettes - TWO SUTURE—caudal margins o First 12g= 6 cassettes > 3 o’CLOCK MARKER o 19g = 7g over 12 g CLINICAL DATA: suture marker at 3 o’clock o 1 cassette per 5g over = 2 more cassettes ― If the chippings weigh 12 g—a minimum of 6 cassettes must be processed > For every 5g over 12 g, one more cassette must be processed EXAMPLE OF SPECIMEN DISSECTION SPECIMEN: skin from back CLINICAL DETAILS: sebaceous cyst > 12 o’clock marker MACROSCOPIC: MICROSCOPIC: ORIENTATION MARKERS (DENOTING MARGINS) ― Aids in the identification and correct orientation of the tissue pieces during embedding ― Assists in the correct placement of the intended surface of the tissue block towards the microtome balded during sectioning ― Distinguishes surgical margins (surgery) from trimming margins A. INKING MAPPING ON A LARGER SCALE - orients specimen component - resection margins - embedding instructions - orientation - distinguish between samples - identify the cut surface - DONE BEFORE BREAD LOAFING - Dry first before applying formalin - India Ink, Silver Nitrate, Various artists pigments - If the specimen is suspected the ink will covered the margin of the specimen - Different color inks are used depends on the dye and different areas - The ink will mark on the actual margin of the slide CASSETTE SIZES STANDARD : 3.0 x 2.5 x 0.4 cm INKING RESECTION MARGINS ― Specimen should not be more than 0.3 cm in thickness > Resection—gi cut na siya ― Tissue cassettes should not bulge when closed SPECIMEN WORKSHEET ― A.k.a “gross worksheet” ― Guides histotechnician in assuring that all blocks are processed ― Must be properly filled up; filled for future reference ― Contains the following; → Accession number → Number of sections and blocks → “comments” column EMBEDDING INSTRUCTION NICKING—indicates laterality > INK DOTS instruct the embdder to embed the tissue a FIXATION certain way ― Crucial step ― Very first step in histopath ― Maintain viability of the tissues ― Process at which the constituent of cells and tissues are being fix in chemical state ― Maintain life like architecture and arrangement of the cell in the tissue ― Can withstand subsequent treatment and various reagents with minimum loss of architecture distortion of decomposition ― It should be fix in order for them not to be damage in the next step of protein processes B. SATURING ― Every steps need chemical or mechanical reagent - indicates anatomical orientation ― We need to fix the tissue for its to withstand for the - used to indicate margins or for orientation treatment after I fix - used variable numbers and/ or colors of suture ― Gina stabilize lang niya ang protein components of tissue - there must be clear description of what the for it to become resistant any changes or damages from sutures indicate in the submission/ request form different reagents applied of our tissues (i.e one suture= cranial margin) HISTOPATHOLOGY ― Important to preserved the tissue after removal from the ― The extent of cross-link formation varies. body (GLUTARALDEHYDE) ― PRIMARY PURPOSE OF FIXATION: preserve the → More effective in cross linking the proteins morphology and chemical integrity of cells → Choice for doing electron microscopy ― SECONDARY PURPOSE OF FIXATION: to hardened and → Stain poorly protect the tissue thus it will be easier for ― Formaldehyde and gultaraldehyde are both formalin histotechnologist or pathologists to cut it solution ― The more the tissue is in the fixative it will hardened due to FACTORS AFFECTING FIXATION detoriation of protein A. FIXATIVE OF CHOICE ― The hardened the tissue easier to cut during in processing ― 10% neutral buffered formalin ― Prevents generation, decomposition, and putrifaction that ― Morphological criteria for diagnosis have been leads to as its common preservation technique established based on FFPEs ― Distortion of tissue after removal of from the body EFFECTS OF FIXTIVES B. pH AND BUFFERS 1. Harden tissues ― (LM) Ph does not affect tissue preservation ― When collecting sample from the body it is fresh ― FORMALIN: pH 6.8-7.2 (acid formaldehyde hematin ― Putting out tissues in fixative it hardened pigment) 2. Act as mordants for certain stains → Below 6.8 the formalin may form acid ― MORDANTS—known as IODINE formaldehyde hematin—clumps or crystal ― Intensify the staining capacity of the primary pigment stain; facilitate staining process ― (EM) pH should match physiological pH 7.2-7.4 ― Known as asintuitors 3. Increase optical differentiation of tissues C. OSMOLALITY ― During microscopic examination ― 400-450 mOsm 4. Render cells insensitive to chemicals ― Measurement of the num