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Cell Bio 2382 - Final Exam Notes.docx

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Biology 2382B
Christopher Brandl

Cell Bio Final Exam Notes Midterm 1 MaterialCell cultures technique to grow cells outside of vivo and under controlled conditions copies physiological conditions such as amount of CO2introduced because cells are used to it 1 Isolationby breaking down the cellcell matrix a Trypsin cleaves the proteins that helps cells stick to each other b EDTA supresses calcium ions that are crucial for sticking onto surfaces2 Supplied with amino acids glucose vitamins mineralsSERUM insulin and growth factorsa Insulin helps take up glucose mitogenetic factorsomething that tells cells to commence cell division 3 Suspension cell culturesroller bottles no time to stick 4Adherent cell culturesstick together plating Phenyl red indicatoryellow is low pH and purple is high pH alkaline Primary cell culturescells directly taken from an organism You can plate these and grow them until they become a confluent monolayer o Takes 9 days o Stops growing because of contact inhibitionTreat with trypsin and transfer to new plate to continue growing o Can only do 50x in humans limitation in labsGerm linescan grow indefinitely and do not show contact inhibition HeLa was the first germ line to be foundCame from cervical carcinoma of Henrietta LacksPicture slide3T3 mouse fibroblast cell that synthesizes cell matrix transformed by Rous sarcoma virus Elongated becomes roundlikeParallel growthrandom growthLoss of contact inhibitionartificially introduced a gene that turned it into a cancer cell Stem Cells Only function is to growAsymmetric division can differentiate into different things OR Self renewal symmetric divisioncontinue producing more stem cells1 Blastocyst is formed basically a ball of cellsa Inside is INNER CELL MASS thing that becomes embryo2 Inner cell mass is the ESC embryonic stem cella Can differentiate into the three germ layers if exposed to the right hormones 3 Less likely to be rejected compared to adult stem cells 4 ESC are then plated on fibroblast cells which are feeder cells responsible for making cell matrix ESC are pluripotentbecome any of the three germ layers but not extraembryonic things such as placentaAdult Stem Cells Most tissues contain Adult Stem Cells Not pluripotentcan only differentiate into what tissue it isUsed to REPAIR and maintain the tissueLocated beside helper cells that give them signals to either differentiate or to self renew Induced Pluripotent Stem Cells iPSCFIBROBLASTSiPSCIntroduce genes to differentiated cells to turn them back into STEM cellso oct4 sox2 klf4 and CMYC characteristic of cancer cellWhen making iPSC you can make cancer cells Cancer Stem CellsStem cells can become cancer cells Also proliferating normal cells rapidly growingCancer stem cells can give rise to any type of cancer anything that grows in a proliferating manner without control apoptosis Lecture 2Microscopy Anton van leeuwenhoekfirst one to use microscope to look at cells bacteria called animcules Blood cells and spermsAchieved a magnification of 200x good at grinding lens Single lens Compoundmicroscope more than one lens increases magnification Condenserfocuses light onto the specimen usually on a glass slip Objective lenscollects the light after it has passed through the specimen magnify100x Eye piece ocularanother lens to magnify10x MagnificationTOTAL of both lenso 40 to 1000x magnification Refractive index ability to bend lightHighermore observableNo refraction by the specimen means all light passes through and its seen as WHITE Any part of the specimen that can slow down light bend it will show up as dark spotsAll the above is bright field microscopy which usually require staining to increase contrast Resolutionability to see two really close objects as two separate objectso 02micrometers apart nothing less thanCONVENTIONAL MICROSCOPESmaller the resolutionbetter Calculationo Resolution D061 lambdaN sin a o Lambdawavelength of light used300700 300 is UV o Nsinanumeric aperture higher is betterNrefractive index of the medium between lens and specimen oilwater a angle of light entering objectiveincrease by bringing the objective lens closer and thus making the cone oflight broadero limit of resolution is 02micrometers200 nmManipulating Phases of light to get contrast Certain parts of cells such as the nucleus have high refractive properties and can slow waves down This results in a shift in the phase of the lighto Nucleus aboutof a wavelength slow down Contrast in microscopy is just darkero Light gets dimmer in a certain area of the cell o INTERFERENCE occurs after the light is recombined at the objective lens How bright light isthe height of the peaks and trophs Interference cancels the trophs and peaks leaving only small bumpsThis is dimmer light bight light has high peaks and low trophsPhase contrast microscopy Does not need to be stained HALO effect SUPER BRIGHT AREAobscure PHASE PLATE that slows down the refracted light additionalwavelength wavelength total1 Annular diaphragm controls the amount of light to pass through2 Condenser lens focuses light onto specimen3 Light refracted by specimen is slowed downwavelength only addition is annularphase plate ONLY THE INNER part of the phase plate slows down light Non impeded light is transferred through the circumference of the phase plateWhen they are combined the contrast is even higher thus the dark spots will be even darker
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