Cell Biology Lab Manual - 5674 Words

Mammalian Cell Culture Lab Manual Student guide of Cell Biology Laboratory Techniques 2012 Jay M. Feldman Cell Biology Professor Lagier 11/26/2012 Mammalian Cell Culture Lab Manual Student guide of Cell Biology Laboratory Techniques 2012 Jay M. Feldman Cell Biology Professor Lagier 11/26/2012 Table of Contents I. Introduction * About the author page 2 * Preface page 3 II. Laboratory Techniques * Cell Culture Technique page 4-5 * Sterile Technique page 6-7 * Media Preparation page 8-9 * Seed cells page 10 * Subculture page 11-12 * Cell count page 13-14 * Cryopreservation page 15 III. Laboratory Experiments * Intentional Contamination page 16-17†¦show more content†¦A. They are immortal B. They are epithelial cells C. They are cancer cells D. All of the above 3) What is an advantage of cell culture? A. Control the physiochemical environment of an experiment B. It is more fun than anti cell culture C. It is easier than anti cell culture D. All of the above Answers: 1) C 2) D 3) A Introduction to Sterile Technique * What is Sterile Technique? Sterile technique is the process of executing cell culture procedures without introducing contaminating microorganisms (bacteria) from the environment * Sterilize vs Sanitize The difference between sanitization and sterilization is that when you sanitize something, you are getting rid of MOST of the live bacteria. It takes about 30 seconds and we perform this using 70% ethanol or Lysol. When we sterilize something, we make free of ALL live bacteria. Sterilization is done with an autoclave or by gas sterilize (this is not done in lab). -Something that is sterilized is also considered sanitized, but if something is sanitized, it is not sterile. * Personal Hygiene By definition, personal hygiene is the act of grooming, cleaning, and looking after our bodies. In cell biology lab, personal hygiene will be necessary in order to keep contaminants from your body from reaching your cells. 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Business Process Reengineering Essay - 1589 Words

Background Business process reengineering has widely become a significant trend in enterprise organizations seeking to innovate and massage business processes. It should come as no surprise that â€Å"over the last decade, numerous organizations have significantly changed their business processes in order to remain competitive in the global market† (Hadaya Pellerin, 2008). The text analyzes business process reengineering as a business process solution for efficiently improving information systems within the context of ERP implementations. The authors suggest that â€Å"BPR is one of the best methods for determining the need to move to an ERP system and set the high level goals and project implementation scope† (Motiwalla Thompson, 2011). In†¦show more content†¦If organizations are in fact seeking to streamline business processes through business process reengineering concepts they must fully understand the BPR methodology. The systematic approach of BPR suggests five st ep methodologies which essentially provide a BPR framework to a successful implementation. The first methodology is the preparation. When analyzing BPR, â€Å"this very first step in BPR is to develop and articulate what is to be accomplished by reengineering, including goals and scope as it relates to BPR† (Motiwalla Thompson, 2011). The as-is process encompasses the functionality of teams and focuses on communicating the visions and goals of the organization. This fundamental process is key in determining the scope that the organization is willing to reengineer contingent upon the successful communication of the individuals involved in the business processes. Thirdly the â€Å"to be† phase focuses on the changes needed to ascertain the outlined objectives of the business processes. This approach can be described as â€Å"a radical new approach to business improvement, with the potential to achieve dramatic improvement in business performance† (Larson Myers, 1997). Essentially if the most important factors that are identified in BPR; the testing and measurement phase of the BPR methodology will facilitate the â€Å"validation of each process necessary to ensure that a step was not missed or that a process wasShow MoreRelatedBusiness Process Reengineering:6310 Words   |  26 PagesBUSINESS PROCESS REENGINEERING: PUTTING THEORY INTO PRACTICE^ BRIAN FITZGERALD AND CIARAN MURPHY Executive Systems Research Centre, University College, Cork, Ireland. ABSTRACT Business Process Reengineering (BPR) advocates the fundamental examination and redesign of business processes, recognising tb-at the legacy of scientific management has been the excessive fragmentation of work practices in organisations today. 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Definition of Lyric Poetry Free Essays

Definition of Lyric Poetry Lyric Poetry consists of a poem, such as a sonnet or an ode, that expresses the thoughts and feelings of the poet. The term lyric is now commonly referred to as the words to a song. Lyric poetry does not tell a story which portrays characters and actions. We will write a custom essay sample on Definition of Lyric Poetry or any similar topic only for you Order Now The lyric poet addresses the reader directly, portraying his or her own feeling, state of mind, and perceptions. â€Å"Italian Sonnet† by James DeFord, written in 1997: Turn back the heart you’ve turned away Give back your kissing breath Leave not my love as you have left The broken hearts of yesterday But wait, be still, don’t lose this way Affection now, for what you guess May be something more, could be less Accept my love, live for today. Written by William Shakespeare: Shall I compare thee to a summer’s day? Thou art more lovely and more temperate. Rough winds do shake the darling buds of May, And summer’s lease hath all too short a date. Sometime too hot the eye of heaven shines, And often is his gold complexion dimmed, And every fair from fair sometime declines, By chance, or nature’s changing course untrimmed. Poem by Emily Dickinson named â€Å"I Felt a Funeral in my Brain. † It describes a person who is going insane, or thinks they are: I felt a Funeral, in my Brain, And Mourners to and fro Kept treading – treading – till it seemed That Sense was breaking through – And when they all were seated, A Service, like a Drum -Kept beating – beating – till I thought My Mind was going numb – And then I heard them lift a Box And creak across my Soul With those same Boots of Lead, again, Then Space – began to toll, As all the Heavens was a Bell, And Being, but an Ear, And I, and Silence, some strange Race Wrecked, solitary, here – And then a Plank in Reason, broke, And I dropped down, and down – And hit a World, at every plunge, And Finished knowing – then – Nonsense Poetry Nonsense poetry is a form of poetry that many people are familiar with, even if they didn’t know they were reading nonsense poetry. The many limericks (both family friendly, and otherwise) that people have read and heard over the years are a form of nonsense poetry. The works of Edward Lear are some of the finest examples of the form. So are the many classic nursery rhymes that we read to our children. Sometimes the language doesn’t make obvious sense and other times the stories being told seem impossible or illogical. Either case can be a technique for writing nonsense poetry. Many of the works of Lewis Carroll are classics of the form. Ronald Dahl is another writer who has entertained us with his strange tales. Words such as silly, strange, bizarre, illogical, whimsical, and fantastic are often used to describe nonsense poems. How to cite Definition of Lyric Poetry, Essay examples

Obligate autotrophs Chemistry and influence of organic compounds Essay Example For Students

Obligate autotrophs: Chemistry and influence of organic compounds Essay In spite of the frequent habitation of compost heaps, nitrate, beds, mud or sewage by nitrifying bacteria and thiobacilli, they are generally incapable of being cultured on a wide range of heterotrophic laboratory media. They may even be inhibited by certain organic nutrients added to the autotrophic medium. Similarly certain diatoms and green or blue-green algae fail to grow on none or only on a very few of the numerous organic compounds tested, and sometimes even then retain a requirement for light and cannot grow heterotrophically in the dark. Numerous metabolic poisons are, not surprisingly, toxic to autotrophs. The existence of facultatively autotrophic relatives of the obligate autotrophs argues against there being any fundamental structural or chemical difference between the two. This view is supported by analysis which shows the usual sugar and amino acid components in polysaccharide and protein of Nitrosomonas, Thiobacillus, Ferrobacillus and blue-green algae. Similarly, the coenzymes, cytochromes, nucleic acids, vitamins and storage products of obligate autotrophs are similar to those of heterotrophs of various types. The endogeneous metabolism in the absence of light or of the inorganic substrates is probably similar to that in heterotrophs. For example, Thiobacillus respires endogenously with an R.Q. of 1 and seems to oxidize a stored polysaccharide synthesized from carbon dioxide. Polyhydroxy-butyric acid is stored by Nitrobacter and by the blue-green alga Chlorogloea10 and presumably provides maintenance energy from its oxidation as it does in Rhodospirillum and Hydrogenomonas. Of course, no growth occurs under such ‘starvation’ conditions, andthere must clearly be some distinction in the obligate autotroph between exogenous organic nutrients and those available within the cell as stored reserve nutrients. Compounds synthesized by autotrophs from carbon dioxide are secreted into the medium to a significant extent. For example, Thiobacillus releases amino acids and phosphatidylinositol into the medium and Ferrobacillus loses pyruvate. These compounds cannot support growth after inorganic substrate exhaustion, but might of course make the environment more favourable for growth: phosphatidylinositol is a wetting agent and may facilitate the attack of elementary sulphur by Thiobacillus. Amino acids in solutionsmight act as chelating agents and provide a bound or ‘buffered’ source of essential trace metals. Such secretion might thus have some selective advantage to these organisms. It should be noted that these compounds probably escape from intact organisms rather than because of lysis, because both T. thiooxidans and Ferrobacillus seem very resistant to lysis. Clearly, no obvious chemical peculiarity is likely to be the cause of obligate autotrophy. Many attempts to find stimulation or inhibition of autotrophic growth by organic nutrients have produced relatively few positive results. Reports of inhibition of growth of nitrifiers by some amino acids and peptone could be due to metal chelation or some other secondary effect and have little significance in considering obligate autotrophy. Similarly growth stimulation by hay infusion and yeast extract could be due to supply of essential trace metals. Such a reason could explain the stimulation of ammonia oxidation in Nitrosomonas by corn steep liquor, for inorganic solids obtained from the liquor are just as effective. Also the stimulation of Nitrobacter by yeast extract may be due partly to molybdenum in the extract, although biotin could be stimulatory also as was found by Krulwich Funk. Certain organic compounds may stimulate autotrophic metabolism. Algal development in the light may be enhanced by the addition of glucose or acetate to the medium. Glucose was slowly used by T. thiooxidans and produced stimulation of sulphur oxidation and growth. T. neapolitanus gives faster growth rates and increased yields when supplemented with amino acids. Nitrosomonas was stimulated by pyruvic acid, which also decreased the lag in autotrophic growth. Photosynthetic growth of Chlorobium thiosulfatophilum was enhanced by acetate but, of course, carbon dioxide, light and sulphide were also essential. These effects could be explained if the stimulatory compounds were used as supplementary carbon sources in a metabolism that was carbon-limited by the rate of carbon-dioxide-fixing reactions or by the synthesis of a particular essential compound such as biotin or a derivative of pyruvate in the examples quoted. Uptake and metabolism of organic compounds Evidence for oxidation of externally added organic nutrients by chemoauto- trophs is relatively scanty but the endogenous respiration of Thiobacillus does seem to be increased by organic acids and by ribose, glycerol and glucose, al- though sometimes this stimulation was slight. Nitrobacter, however, is able to oxidize formate quite rapidly and the oxidation seems to involve the same cytochrome system as that involved in nitrite oxidation although the pH optima for the oxidation of the two substrates are different. Formate oxidation is apparently coupled to carbon dioxide fixation and may even allow a very low rate of growth. A Worn Path Theme EssayHowever, Thiobacillus denitrificans is freely permeable to glycerol, on which it cannot grow, and Nitrobacter similarly accumulates materials which do not support growth. It is noteworthy that Thiobacillus neapolitanus assimilates virtually none of any acetate or amino acids supplied to it, unless it simultaneously oxidizes thiosulphate. Similarly acetate assimilation by Anacystis nidulans21 is light-dependent. This may indicate that permeation by the nutrients occurred only if a source of energy were available. Clearly, however, permeability is not the explanation for all autotrophy. 2. Energy coupling Perhaps the most attractive explanation is to believe that even if the organism is permeable to the nutrient it is unable to grow on it, because of an inability to obtain sufficient energy from its oxidation. Such inability might result from a failure to oxidize the compounds or from a lack of the systems known in heterotrophs for the trapping of energy from such oxidations. Many autotrophs do not oxidize organic compounds to any significant extent and even if they do stimulate respiration they do not support growth. Lewin showed that even complete oxidation of acetate by Chlamydomonas dysosmos was possible but no growth occurred. The fact that oxidations may occur, shows that there is permeation into the cells, and thus indicates that the oxidations may not yield energy to the organism. Lewin in fact simulated the responses of a strictly autotrophic Chlamydomonas to acetate by treating a wild type strain with the energy-uncoupler 2,4-dinitrophenol. Hempfling ; Vishniac found that although Thiobacillus extracts oxidized NADH its oxidation did not support ATP synthesis. If intact organisms behave in the same way, NAD-linked oxidations would not support growth, unless the cytochrome systems believed to be involved in sulphur oxidations were also involved in NADH oxidation. The critical factor in Nitrobacter growing with formate may well be that the energy trapping system is the same as with nitrite. The main function of light energy or chemolithotrophic energy in supporting organic assimilations by Chlorobium, blue-green algae and thiobacilli is almost certainly in activating the compounds and providing sufficient energy for synthetic reactions. However, if this were the sole explanation for dependence on light or specific inorganic respiratory substrates, an ‘obligate autotroph might be able to grow on organic compounds in the absence of carbon dioxide provided that the particular energy source is also supplied. However, experiments with Chlamydomonas, Nitrobacter and thiobacilli failed to obtain such growth when even complex mixtures of organic compounds, including extracts of the organisms themselves, were provided. Thiobacillus neapolitanus, for example, could not develop without C02 but with thiosulphate when supplied with hexoses, pentoses, yeast extract and amino acids, although normal development occurred in the presence of C02. Butler ; Umbreit16 obtained growth of T. thiooxidans on glucose with sulphur in the absence of C02. This result does not mean, however, that total cell synthesis produced from glucose directly, for Suzuki27 showed that this organism could incorporate glucose into aminoacids and other cell components during sulphur oxidation, but that it was also oxidized (possibly by both Embden-Meyerhof-Parnas and pentose-phosphatecycle mechanisms) to liberate carbon dioxide. Consequently much of the cell biosynthesis observed by Butler ; Umbreit could have depended on this liberated C02, which would be providing physiologically ‘normal’ conditions. With Chlorobium and blue-green algae the light-dependent incorporation of organic compounds also depends on simultaneous carbon dioxide supply. Growth could not thus occur on organic nutrients alone, but in considering the C02 requirement of autotrophs one must recall that it is an essential metabolite to most, if not all, heterotrophs also. The large amounts of glucose and acetate carbon (32% and 18% of the dry weight) assimilated by the Anabaena of Carr ; Pearce28 might similarly have been taken up in part as C02 previously released as oxidation products of, say, an operative tricarboxylic acid cycle.