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Relay Race Trivia: Biology & Lab Basics ETEAMS 2014 By: Kelly Correia & Kaitlyn Schroeder-Spain 1 Slide 2 Introduction Discuss topics and information relevant to ecotoxicology project and basic/general biology Expect this material to show-up in the trivia section of the relay race!! Topics include: Common glassware & use Laboratory equipment & use Anatomy: Male vs. Female Blue Crabs Basic Biology Review 2 Slide 3 Volumetric Flask Aka a measuring flask Usually pear shaped Used to obtain a precise & accurate volume of a solution Mixing: insert cap, secure with parafilm, and invert Range of sizes, commonly 25 mL 2 L 3 Slide 4 Petri Dish Also called a cell culture dish. Consists of a plate + lid Commonly used in microbiology laboratories and courses Microbiological cultures can be grown in petri dishes of differing sizes Often have a thin layer of growth medium, ex: agar 4 Slide 5 Beakers Commonly used in laboratories for stirring, mixing and heating liquids Not used to measure precise volumes Large range of sizes: milliliters to liters 5 Slide 6 Graduated Cylinder Used to measure solution volume More accurate and precise method of measurement than beakers Less accurate and precise than volumetric glassware or volumetric pipettes Meniscus: the curved upper surface of a liquid in a tube. Bottom of Meniscus is used for measurements; should sit directly on the mark 20 mL 6 Slide 7 Pipettes Several types, but all used to measure relatively small amounts of solutions and samples. Some are disposable, others only partly. Pasteur Pipette -Blubs may be reused, or not -Glass is disposable -Crude, small measurements Disposable pipettes -similar to droppers -Plastic - Often used only once - For small samples & crude measurements Volumetric Pipette (+ bulb) -Blubs reusable -Glass disposable -marked like graduate cylinder 7 Slide 8 Micropipettes Non-disposable; provide the highest degree of precision and accuracy Can be single-channel or multi-channel Pipette tips: Are disposable Can be sterile (usually boxed) or non- sterile (usually in a bag) Come in a variety of sizes, depending on pipette & volume of sample Multichannel pipette 8 Slide 9 Each brand is slightly different, but also similar Sample Volume: will determine the pipette and tip size used (Table 1) Change volume: turn plunger OR a separate knob; often youll hear a clicking sound Eject tips: push eject button Pipetting session will provide actual training Micropipette Use: Basics Table 1. Example Pipette &Tip Sizes, Volumes Pipette typeVolumes (L)Tip color P100.5 10white P202 20yellow P20020 200yellow P1000200 1000blue 9 Slide 10 How To Read a Micropipette 1.Confirm range: labeled on plunger & often one side Range will determine decimal places, do not rely on colors for determination Note: knobs turn beyond their range, and can break P1000 Panel View P1000 Values P200 Panel View P200 Values P20 Panel View P20 Values 11000s2100s210s 0100s010s01s 010s01s0decimal = 1000 L (1 mL) = 200 L (0.2 mL) = 20 L (0.020 mL) 10 Slide 11 How To Read a Micropipette, Example # 2 P1000 Panel View P1000 Values P200 Panel View P200 Values P20 Panel View P20 Values 01000s1100s110s 5100s810s51s 010s51s2decimal = ____ L (0.5 mL) = ____ L (0.185 mL) = ____ L (0.0152 mL) 11 Slide 12 Conversions (self study/Reference) Expect to convert between different volumes of solutions Practice and review will be covered during pipetting and graduated cylinder sessions 12 Slide 13 Stir Bar & Stir Plates Both are used to mix a solution thoroughly Help avoid and/or assist manual stirring Stir bars are magnetic, plastic and reusable; come in a variety of sizes and shapes Stir plate is also magnetic, to move stir bar Stir bar is gently dropped into a solution and placed on stir plate; plates have several speeds Use a Stir Bar Stick or Retriever to remove stir bar from solution (also magnetic) Stir bars (above) Stir Bar Stick/Retriever (right) Stir plates 13 Slide 14 Hot Plate & Stirrer/Hot Plate Combo Hot Plate (alone) Used to heat a solution, often to assist with mixing (increase solubility) Turn dial to change heat level Stirrer/Hot Plate (combo) Used to mix AND heat a solutions, Turn separate dials to change heat level and stirring speed 14 Slide 15 Used to separate samples by weight, size Can be micro (table top) or very large As rotor spins heavier particles separate on bottom of centrifuge tube Produces a supernatant & a pellet, which can be separated, re-suspended (pellet), and spun at higher speeds if necessary Centrifuge speed varies with size of centrifuge rotor and can vary with overall size Desired molecules, enzymes, etc. will determine speed necessary for separation Centrifuges 15 Slide 16 Seawater & Salinity Salinity units Oceanic World avg. 35.5 PSU PSU = practical salinity scale Estimate of ionic content 1 PSU = 1 g/kg Old method: expressed as %, or ppt (parts per trillion). Known as Knudsen salinities. Laguna Madre = unique! Hypersaline Lagoon Blue crab tanks are kept at 18-20 PSU Measure salinity using the refractometer What salinity is the image showing on the left? 16 Slide 17 17 Slide 18 Crab Sex Identification Abdominal apron, or flap, is shaped differently for & : upside down U = mature; upside down v immature : pillar shape, & slender entire life cycle Claws different colors Immature , : claws appear greenish/white, with hot pink dots Mature : claws are bright orange / red Immature : claws are blue, sometimes greenish-blue 18 Slide 19 Female Blue Crabs (FYI) Abdominal apron, or flap, is used to carry eggs 19 Slide 20 Plant VS Animal Cell (Major Differences) Animal cells: no cell wall; can have flagella, cilia & lysosomes Plants: cell wall, chloroplasts 20 Slide 21 DNA Structure Genetic information for all living things Humans have 46 chromosomes, or 23 pairs 1 pair sex chromosomes 22 pairs = autosomes Meiosis 24,000 genes ( = 2% of entire DNA) 95 % identical to chimpanzee ~ 50 % identical to bananas 21 Slide 22 RNA Structure 22 Slide 23 5 Kingdoms of all living things Monera (includes Eubacteria and Archeobacteria) Protista Fungi Plantae Animalia 23 Slide 24 Stages of Mitosis (IPMAT) *Be able to put images of Mitosis in order for relay race 24 Slide 25 Structure of a water molecule Be able to draw or create a water molecule, including charges and bonds Water = 2 Hydrogen atoms covalently bonded with Oxygen Water is bipolar because: electronegativity of Oxygen + electropositivity of 2 Hydrogen atoms Bonds between water molecules = hydrogen bonds 25 Slide 26 1.Carl Linnaeus 2.Charles Darwin 3.Louis Pasteur 4.Gregor Mendel 5.Watson and Crick (+Franklin) 6.Rachel Carson 26 Slide 27 Carl Linnaeus (1707 1778) father of biological systematics and nomenclature He invented the modern classification system of living organisms Binomial nomenclature e.g., blue crabs = Callinectes sapidus 27 Slide 28 Charles Darwin (1809 1882) Naturalist and Geologist Voyage of the Beagle (1831 1836) Origin of Species (1859) Theory of Evolution all species of life have descended over time from common ancestors 28 Slide 29 Louis Pasteur (1822-1895) Chemists and microbiologist Created first vaccinations for rabies and anthrax Primary founder of microbiology (along with Cohn & Koch) Proved that most infectious diseases are caused by micro- organisms Known as the germ theory of disease Developed pasteurization process Prevent milk and wine from becoming contaminated with bacteria 29 Slide 30 Gregor Mendel (1822 1884) Most known for his famous hereditary experiments with pea plants (1856-1853) Mendelian Laws of Inheritance, work with peas and flowers; studied phenotypes of several generations Discovered the basics of Genetics by crossing/mating peas with different physical traits 30 Slide 31 James Watson (1928 - ) and Francis Crick (1916-2004) + Rosalind Franklin (1920 -1958) Watson (left) & Crick (Right) worked at Cambridge University Discovered & published DNA structure (1953) Attended Franklins lecture Rosalind, with others (Wilkins), worked at Kings College She studied x-ray diffraction of DNA, helped ID phosphate bone and helix structure 1962: Watson, Crick, and Wilkins won the Nobel Prize for physiology/medicine Franklin died in 1958; no posthumous prizes 31 Slide 32 Rachel Carson (1907-1964) Author, Silent Spring (1963) and several other books Focused on effects of DDT and other pesticides One of the most influential people of the 20 th century Famous for advancing the global environmental movement 32