Laboratory Safety & Instrumentation 2021-2022 PDF

Laboratory Safety, Instrumentation, and Quality Management in Histopathologic Laboratory LAB 2021 – 2022 HISTOPATHOLOGY/ CYTOLOGIC TECHNIQUES 2nd Semest...

Laboratory Safety, Instrumentation, and Quality Management in Histopathologic Laboratory LAB 2021 – 2022 HISTOPATHOLOGY/ CYTOLOGIC TECHNIQUES 2nd Semester Instructor: Kimberly Pulga, RMT, MPH WEEK 1 HPCT311 LAB Date: February 15, 2022 - Retention period – refers to how many years, TOPIC OUTLINE months, weeks, and days that the specimen will be A. Laboratory Safety stored. I. Histopathology - Overdue – disposal of the specimen II. Risk Management III. Risk Management and Safety in the II. Risk Management Laboratory ❖ In the laboratory, we are assessing the risk and hazards IV. Types of Hazards that are present A. Chemical Hazards ❖ Process of ensuring and maintaining personal as well as I. Labelling environmental health and safety in the laboratory. II. Different Types of ❖ The first step is to identify electrical, mechanical, and Chemicals biological that can potentially cause harm in the laboratory. A. Physical Hazards ❖ Electrical – in any workplace not just in the laboratory B. Biological Hazards (school, house, etc.) B. Instrumentation ❖ Mechanical – instruments in the histopathology laboratory I. Use and Care of Microscope (microtome, microscope, for cutting/slicing tissues) II. Compound Microscope ❖ Biological – specimens that can be the source of A. Eyepiece pathologic microorganisms. They can potentially cause B. Objective lens harm C. Bright field microscope ❖ An inventory of chemical reagents must be on hand and D. Dark field illumination obsolete chemicals should be routinely disposed of. E. Phase-contrast microscopy ❖ *In the histopathology section, madami kayong reagents polarized microscopy na makikita doon such as decalcifying agents, clearing F. fluorescence microscopy agents etc. G. electron microscopy I. Transmission electron III. Risk Management and Safety in the Laboratory microscope (TEM) Protect the hands and forearms by wearing either gloves II. Scanning electron microscope (SEM) and a laboratory coat or suitable long gloves to avoid C. Quality Management contact of the toxic material with the skin. Wash hands I. Laboratory Quality and Management frequently throughout the day and before leaving the lab. System Procedures involving volatile toxic substances and those II. Continuing quality improvement involving solid or liquid toxic substances that may result in the generation of aerosols should be conducted in a fume hood or other suitable containment device. A. Laboratory Safety The laboratory workplace should be well-ventilated, clean and organized. I. Histopathology Smoking, sleeping, eating and drinking are prohibited in ❖ Art of analyzing and interpreting the shapes, sizes, and the laboratory. architectural patterns of cells and tissues within a given Do not store food and drinks in laboratory refrigerators. specific clinical background. Do not wear shorts, sandals, or open-toed shoes in ❖ “Histo” – the study of tissue in a microscopic level, laboratory. Minors or personal pets are not permitted in the ❖ “pathology” – a study of the diseases laboratory. Secure any dangling jewelry, restrain loose ❖ Slides are prepared inside the histopathology laboratory clothing, and tie back long hair that might get caught in and it is checked by pathologists equipment before starting work. ❖ Activities: Use of cell phones and music headphones should be I. Pre-analytical avoided while working in the lab. They can be distracting - specimen collection and can increase the potential for an accident to occur. - accession number They can also become contaminated if handled while II. Analytical working with hazardous materials. - checking for the tissue processing Every instrument used in the laboratory should meet - reading of prepared slides is done by a specialist electrical safety specifications and have written physician instructions regarding its use. - make sure that relevant data are provided Eyewash station, safety shower and first aid kits should be III. Post-analytical standard facilities in a laboratory. Fire extinguishers, - Preparation and delivery of the report to the emergency shower systems, emergency eye washers, first requesting physician aid, emergency blankets, and hoods must be checked - Storage of the request forms *tinatago ang mga monthly. request forms hindi tinatapon To avoid the unnecessary purchase of chemical materials, - Storing of the reported specimen a detailed list of chemical materials must be prepared. - The slide being used will not be disposed of instead Only a minimum amount of volatile chemicals must be the reported specimen will be stored also called kept in the laboratory. Archiving or Storing of the reported specimen - The storage will be depending on the type of Chemical material should be stored and safely secured specimen that you have *Naka-depende yung storage where there is sufficient ventilation. Combustible chemical sa slide na nagawa ng medtech. material must be stored in a heat-resistant cabinet. Acids and bases must be separately stored. C. Caldito & J. Cotelo - TRANSCRIBERS  Laboratory Safety, Instrumentation, and Quality Management in Histopathologic Laboratory LAB 2021 – 2022 HISTOPATHOLOGY/ CYTOLOGIC TECHNIQUES 2nd Semester Instructor: Kimberly Pulga, RMT, MPH WEEK 1 HPCT311 LAB Date: February 15, 2022 Every chemical compound used in the laboratory should din. Walang enough knowledge about the chemicals that have a materials safety data sheet on file that specifies the we are handling nature, toxicity, and safety precautions to be taken when ❖ cleaning agents, disinfectant drugs, anesthetic gases, handling the compound. solvents, paints, and compressed gases. All chemical material must be labeled with the name, ❖ *so sa histopathology, yung mga reagents lang naman characteristics, danger level, and precautionary measures. talaga ang nandoon such as chloroform, formalin, Laboratories must have available appropriate protective benzene, formaldehyde, etc. gears for all individuals: safety devices, goggles, gloves, ❖ The “lab standard” applies to the laboratory use of lab coats, and face-shields. chemicals and mandates written in the standard The laboratory must have a method for disposal of operating procedure (SOPs) that address the particular hazardous wastes. Collect and seal absorbed material into hazards and precautions required for safe use. labelled containers for disposal. Tissues that are collected ❖ Terms: should be stored in formalin and may be disposed by PELs - permissible exposure limit incineration or by putting them through a "tissue grinder" TLVs - threshold limit value attached to a large sink (similar to a large garbage OELs - occupational exposure limits disposal unit). Used chemicals must not be released into ❖ *These are just some of the terms to define the maximum soil, drains and waterways. Use an absorbent such as allowable airborne concentration of a certain chemical, sand, “kitty litter” or a commercial product to collect spills whether vapor man yan, fume or dust to which a worker and contain spread. may be expose One must always be cautious when handling electrical appliances and must be aware of the location of safety I. Labeling - Every chemical should be labeled with certain devices (fire extinguisher, emergency shower system). basic information, including: Extinguishers with water, carbon dioxide, dry chemical o Chemical Name powder or foam are all suitable depending on other - if mixture, the names of all ingredients should products involved in a fire. Fire safety procedures should be included be posted. There must not be any obstacle in the vicinity o Manufacturer’s name and address of the laboratory door. - if purchased commercially Avoid handling the sharp ends of instruments. Use forceps - If ikaw yung gumawa ng reagent or nag halo- or other tools to remove sharp instruments from baskets halo niyan dapat nakalagay dun ex. Pulga and autoclaves. Workers should use appropriate hand o Date purchased or made protection when hands are exposed to hazards such as - Date purchased kapag binili commercially cuts, lacerations or thermal burns. - Date made kapag ikaw mismo yung gumawa ng Laboratory accidents must be documented and reagent (ex.Pulga 2215) investigated with incident reports and industrial accident o Expiration date reports. Obtain medical advice (first aid officer, doctor, o Hazard warning poisons information center, ambulance) immediately if - Dapat icheck yung diamond shape symbol or major exposure occurs. NFPA One of the most common accidents in the laboratory - Colors seen in NFPA are blue, red, yellow and involves cutting of one's finger or hand on microtome white knives. (Gregorios) ❖ very important na ginagawa dapat sa mga chemicals sa *Cutting of one’s finger or hand kasi we are handling histopathologic lab or even in general, the labeling of the microtome kasi doon tayo nag cu-cut ng tissues sample chemical natin, and sobrang matatalim yung knives non. Minsan matigas kasi yung tissues. *kwento muna si ma’am ng II. Different types of chemicals include: past experience nya. Mostly daw boys ang gumagawa. o Irritants – reversible inflammatory effect - At the site of contact with the living tissue IV. Types of Hazards especially our skin, so dapat kapag nagkaroon A. Chemical – dealing with a lot of reagents in the histopath lab ng exposure, hugasan or flash with water (formalin, etc.) - If sa eyes, we should go to the eye wash station B. Physical – common in a general setting pero mas maganda if you are wearing your C. Biological – since we are receiving specimens and it can be goggles whilde doing any tissue processing in a source of potential microorganisms (surgically removed the histopathology laboratory kasi masakit sa organs) mata and it can go to the respiratory passage - Total Abdominal Hysterectomy and Bilateral o Corrosive – destruction or irreversible alteration Salpingo-Oophorectomy (TAH-BSO) – a - it can destroy certain inanimate surfaces like surgical removal of uterus, cervix and fallopian metals. So dahan-dahan kapag corrosive ang tubes, and ovaries hawak o Sensitizers – allergic reactions - Hindi lang sa mga hypersensitive individuals, A. Chemical Hazards basta sensitizers ang na encounter talagang ❖ Reagents that we are handling in the histopathology lab magkakaroon ng allergy and kapag mataas ang ❖ *Dito, pag meron kang potential exposure sa chemical exposure nang worker hazards, pwedeng magkaroon ng chemical spill, o Carcinogens – tumors accidents, worst is the exposure pag pinag-combine yung - Substances that induces tumor, not just in mga dapat hindi dapat magkakatabi animals but also in humans especially in ❖ *Nangyayri to kapag poor yung storage at yung paggamit prolonged exposure to chemicals and later on it C. Caldito & J. Cotelo - TRANSCRIBERS  Laboratory Safety, Instrumentation, and Quality Management in Histopathologic Laboratory LAB 2021 – 2022 HISTOPATHOLOGY/ CYTOLOGIC TECHNIQUES 2nd Semester Instructor: Kimberly Pulga, RMT, MPH WEEK 1 HPCT311 LAB Date: February 15, 2022 can lead to a cancer most specially in staining, cutting, at kung - Example: chloroform, chromic acid, formalin, nasama ba yung part ng tissue na dapat ay formaldehyde, carcinogenic dyes, potassium makita ng pathologist chromate, auramine, basic fuchsin and even the - dyes that are derived from benzidine such as ❖ Must accomplish the following: congo red, diaminobenzidine 1. it must magnify the object o Toxic materials – capable of causing death by ingestion, - That is the factor by which an image will appear inhalation, skin contact, etc. to be enlarged. - It causes death by ingestion and inhalation but - To know the cellular abnormalities at certain specified concentration 2. it must resolve the details of the object - Example: methanol, chromic acid, osmium - Resolving power tetroxide or even uranyl nitrate - So dapat the microscope that you are using has the ability to distinguish between small objects B. Physical Hazards that are close together. So aalamin yung ❖ slips and falls from working in wet resolving power ng microscope na gamit ninyo locations and the ergonomic - Resolving means the ability to separate or to hazards of lifting, pushing, pulling, distinguish two small objects that are close and repetitive tasks. Other physical together hazards often unnoticed are - So kapag masyadong maliliit na yung objects electrical, mechanical, and thermal. yung dalawang objects na yun ang magiging Ignoring these can have potentially tingin niyo na lang ay isa. serious consequences. 3. it must make these details visible. - If you are about to lift something specially when it is heavy object make sure to bend your knees II. Compound Microscope kasi mabibigla yung spinal cord ❖ Also called as Bright Field Microscope ❖ Many operations in the lab can result in lab workers - which contains own light source assuming sustained or repetitive awkward postures such at its base. So, the specimen, iniilawan siya as looking at slides on a microscope for extended periods below and they are viewed from above. Parts of Compound Microscope: C. Biological Hazards Ocular (Eye piece) ❖ Anything that can cause disease in human regardless of Body or Tube their source so since we are in the histopathology, we Course focusing knob must again in general rule must consider all specimen Fine focusing knob that we are receiving as potential source of harmful Objective lens microorganisms. Movable stage- it is the platform where the slide is place ❖ Include infectious agents and their toxins as well as for the examination contaminated solutions, specimens, or objects. Substage- it is under the stage and holds the condenser ❖ Allergens, are one of the most important health hazards, and your diaphragm. yet they are frequently overlooked Mechanical Stage- it permits the movement of the stage ❖ Molds and fungi produce and release millions of spores while holding the slide in the face and focus. small enough to be air, water, or insect-borne which may Condenser lenses have negative effects on human health including allergic Field (iris) diaphragm reactions, asthma, and other respiratory problems. Mirror & light source Base- it provides the support for the microscope. It B. INSTRUMENTATION should be large and solid enough para mallow natin magstand yung microscope by itself. I. Use and Care of Microscope Arm- its supports and holds your magnifying and adjustment system. We also used this as handle for ❖ Microscope is one piece of equipment that is used by carrying the microscope. both the pathologist and the histotechnologist ❖ pathologist examines the slide under the microscope to identify a disease process or an abnormality that will A. EYEPIECE/OCULAR directly affect the patient's treatment. ❖ Forms magnified virtual & erect - Specialist pathologist are the one who examines images the slides under the microscope for the Types: identification of the diseases and also to check if a) Monocular (Huygenian) there are cellular abnormalities - One eyepiece when you ❖ Histotechnologist examines the same slide microscopically are viewing the specimen. It is also for quality control to determine whether all technical very light weight and inexpensive. processes are done properly and if a slide of diagnostic b) Binocular (Ramsden) quality has been achieved. - Two eyepiece. They are convenient and - They are the ones who prepare the slides for comfortable to use. observation and also at the same time they also - Common choice examine the same slide microscopically for c) Trinocular (Compensating) quality control - Have a third eyepiece - To check if all the technical processes are done - The use of third eyepiece is for another para C. Caldito & J. Cotelo - TRANSCRIBERS  Laboratory Safety, Instrumentation, and Quality Management in Histopathologic Laboratory LAB 2021 – 2022 HISTOPATHOLOGY/ CYTOLOGIC TECHNIQUES 2nd Semester Instructor: Kimberly Pulga, RMT, MPH WEEK 1 HPCT311 LAB Date: February 15, 2022 dalawa yung pwedeng magtingin sa specimen or tatlo para tag-iisa for each person. ❖ The light microscopy of phase contrast enhances the B. OBJECTIVE LENS contrast of transparent and colorless object. It is also ⚫ Mounted on nose piece influence the optical path of light kaya ganto yung If forms magnified real image nakikita niyo (referring to letter b picture) Magnification of objective = Optical Tube Length F. POLARIZED MICROSCOPY Focal length (the distance between the outer lens of the ❖ contrast-enhancing technique that improves the quality of objective and the cover glass of slide under the examination. the image obtained with birefringent materials when Scan – 4x X 10x eyepiece = 40x compared to other techniques such as bright field Low power – 10x X 10x eyepiece = 100x microscopy, phase contrast microscopy and darkfield High power – 40x X 10x eyepiece = 400x microscopy. Oil immersion – 100x X 10x eyepiece = 1000x ❖ In here, this is design to examine specimens that are visible primarily due to their optically anisotropic character. C. BRIGHT FIELD MICROSCOPE ❖ Two essential components: ❖ Generally used in compound microscopes, Polarizer where light is either passed through or - This is situated reflected off, a specimen below the specimen ❖ a specimen is placed on the stage of the stage. Nakafixed to in microscope and incandescent light from the left to right, east to west microscope’s light source is aimed at a lens direction. beneath the specimen. - Pero may iba na rotatable naman siya D. DARK FIELD ILLUMINATION 360 degrees. Yung iba ❖ technique used to observe unstained and transparent naman is east to west, samples causing them to be clearly visible and appear left to right na direction. brightly lit against a dark, almost purely black Analyzer background. - Located siya above the objectives. - This is usually aligned in north-south but again, ❖ Ginagawa dito sa dark field microscope, binablock dito rotatable rin siya in some microscope 360 yung central light gamit yung condenser para yung mga degrees. oblique rays lang yung maghit sa object. - It can be moved in and out of the light path as required. G. FLUORESCENCE MICROSCOPY ❖ 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 cameras. ❖ When certain compounds are illuminated with high energy light, they emit light of a lower frequency ❖ So, specimen is stained with fluorescent dye then after that, when we will examine it under the microscope using your UV light and makikita niyo na magfloulfourish yan and then you can see there a visible radiation. E. PHASE CONTRAST MICROSCOPY ❖ So, may ultraviolet light and then you will see a bright ❖ type of light microscopy that enhances contrasts of object against the dark background. transparent and colorless objects by influencing the optical path of light ❖ phase-contrast microscope is able to show components in a cell or bacteria which would be very difficult to see in an ordinary light or bright field microscope. ❖ We used this to have a better visualization of internal cellular components. ❖ Pwede rin ito for the diagnosis of tumor cells and also examination of the growth and behavior of wide variety ng living cells in the cell culture. C. Caldito & J. Cotelo - TRANSCRIBERS  Laboratory Safety, Instrumentation, and Quality Management in Histopathologic Laboratory LAB 2021 – 2022 HISTOPATHOLOGY/ CYTOLOGIC TECHNIQUES 2nd Semester Instructor: Kimberly Pulga, RMT, MPH WEEK 1 HPCT311 LAB Date: February 15, 2022 o United Kingdom National External Quality Assessment Service (UK NEQAS) ❖ Quality Assurance – H. ELECTRON MICROSCOPY ❖ two distinct systems: ❖ Electron Microscopes uses a beam selective system: stained preparations from of highly energetic electrons to examine departmental archival records are used to assess the objects on a very fine scale. quality of staining. ❖ This examination can yield the info Distributive system: participating laboratories are asked about to stain sections that have been submitted by the Morphology scheme organizer Composition II. Continuing Quality Improvement ❖ The examination can yield info about the morphology and ❖ system is used to approach, evaluate and identify compositions of the tissues or whatever that is you viewing opportunities to improve quality before problems occur under the electron microscope through evaluation of all systems/processes in the laboratory a. TRANSMISSION ELECTRON MICROSCOPE (TEM) ❖ GOAL: improve potential care & safety through recognition ❖ Stream of electrons is formed of potential problems/errors before they can occur ❖ Accelerated using a positive ❖ electrical potential ❖ Focused by metallic aperture and electro magnets ❖ Interactions occur inside the irradiated sample which are detected and transformed into an image ❖ Projector lens forms image on the fluorescent viewing screen ❖ 2D image ❖ Magnification 10,000X to 100,000 X ❖ It works on the same principle as an optical microscope. ❖ So ang ginagamit lang dito is electron instead of the light Book References: and electro magnets instead of the glass lenses. So ayun References: pinagkaiba nila. Bruce-Gregorios, Jocelyn H. Histopathologic Techniques, ❖ We are using this electron microscope for thin slices of 2017 Lo, Raymundo, et al. Basic Histopathologic specimen Techniques, 2015 Adyanthaya, Soniya, et al. Quality and safety aspects in b. SCANNING ELECTRON MICROSCOPE (SEM) histopathology laboratory, 2013 ❖ 3D image C. QUALITY MANAGEMENT I. Laboratory Quality and Management System ❖ Quality - degree to which healthcare services strive to provide accurate desired outcomes for patients and are consistent with current professional knowledge ❖ Safety – “freedom from accidental injury” ❖ Quality Control - system of routine technical activities ❖ Quality Assurance - planned system of review procedures conducted by personnel not directly involved in the laboratory process data of QC provides the data for QA Quality assessment programs o College of American Pathologists (CAP) C. Caldito & J. Cotelo - TRANSCRIBERS