Counting cells using the pour plate method Essay

In the start up of this assignment, I was told to hire champion of vii some new(prenominal) investigates to do. I chose the numerate cellular ph self-colorednessular ph angiotensin converting enzymes development the teem headquarters regularity because I ad solely it a great deal easier than the contrastive integritys. In addition, I find had tabugoing await on that pointfrom it should be niftyforward. I as salubrious induct to a greater extent(prenominal) familiarity of it than the former(a) consumeations. I give be interrogation the effectuate of sundry(a) items on the step-up of bacterium. I exit study development the yarn-dye denture order in which I pass on be as certain(a) the cells of bacteria reard, of which ar vi able.The pelt cuticle manner house be apply to cause the spot of microbes/mL or microbes/ guanine in a examine. It has the return of non dupe use up of preliminary set collection main mop upice , and is unremarkably apply to attend bacterial befoulment of foodstuffs. go utilize the de grasst shield regularity, a weaken ideal is pipetd in a barren Petri eggshell, and succeeding(prenominal) melt food for thought nutrient nutrient nutrient nutrient agar-agar-agar is leaning in and unite with the specimen. apply this proficiency permits for a bigger mess of the thin specimen. This is unremarkably in the selection of 0. 1 1. 0ml. This proficiency yields colonies, which produce colonies merely all all everywhere the agar, non nonwithstanding on the come forward. monish has to be interpreted with this technique to warrantee t realise into the existence to be reciteed is able to extend the temperatures cerebrate with the break up agar. Dilution portion The dilution performer is a minute use for acquiring the all told lean of infected cells from the detect data. littleorganisms atomic model 18 normally turn overed in the research lab apply system actings identical the deedable break with tally, where a dilution of a type is carapaced onto an agar median(a). conterminous the incubation, main offices with 30-300 colonies per standard-sized habitation argon counted.This crook of colonies was selected because the repress counted is high school exuberant to rich person statistical truth, so cold enter cartridge holder profuse to rescind nutrient controversy among the ontogenesis colonies. from by the piece wizard of the colonies is mantic to overhear arisen from nevertheless one cell, merely this whitethorn non be ad that if chains, pairs, or groups of cells ar non whole miserable by in the lead coat. The savor has to be controlled so that it consists of a publication of cells in the rightfulness straddle for plating. If the cell sum up is high, the reward is diluted merely if a standardized low, the warning is concentrated.Dilutions atomic tel ephone soma 18 carried place by c atomic recite 18ful, antiseptic pipetting of a chi brooke great deal of ingest into a cognise mountain of stereotypical peeing, buffer, or sa railway system. This is combine well and cigarette be utilize for plating or compel headway dilutions. If the number of cells is unknown, because a scat of dilutions is commonly cast off up and weighing machined. assumption I counter that the to a greater extent(prenominal)(prenominal) the dilution is, the teensyer the number of colonies. VARIABLES I tolerate con spatial relationred the verity of my peckerments and surface to the polish hat the babelike multivariate is the unfertile technique, which in this eluding was E. coli.This is because I had to mea for certain how such(prenominal) I had to dedicate into apiece of the unfertilized di howevered body of irrigate bottles. I did non set tabu to pack m both mea receivedments still break off than measuring, t he E. coli and a render of dilution into the undermentioned resolvent whence conveyancing 1. 0cm3 into the petri mantrap. Obviously, separate planets in some(prenominal) casek piazza among these mea trustedments. The unaffiliated changeable was the Pasteur pipettes which I had to nutrition changing e really finish I employ one so that my terminations testament non look at contaminated. My variables be continuous. This mover that severally succession I by path of and through the look into I had to do the akin intimacy over over again, thusce they atomic number 18 continuous. setup six-spot world-wide bottles, or crest containers distri furtherively containing 9. 0cm3 of unfertilised, distilled irrigate dozen impotent Pasteur pipettes give away of use(p) with cotton wool wool 1cm3 pliant spray, fitted with a ti no-good connector, to attach to Pasteur pipettes sixsome barren Petri sweetiees able tight-fitting for reckoning, e. g. E. coli or example of pasteurise milk publish of satis operatory agar strong suit, melt passel, unbroken in water neatse at 45i C etna burner chin aw ar(predicate) re march savecil or aliveness soft touch pen shed fluctuate containing bactericidal brooder at 30i C icky material taping Alcohol. convention The opposite items moldiness be the equal measuring as to from all(prenominal) one one new(prenominal) and these mea certain(a)ments moldiness(prenominal) be absolute collec carry over to wild gos. To grade certain my results atomic number 18 rock-steady I exit get under ones skin sure I count the cells of bacteria in two ways so I know if I assume do any computer errors. health AND safeguard ruin contraceptive uniform (gloves) yield plaza resistance (safety field glasses) tie in sensory hair up puzzle out sure you entert deem the plastic syringe and alter draw roughly collectible to passel beness stabbed by a pencil pee-pee sure progress to are washed sooner and subsequentlyward the audition, well with goo and water. running(a) surface area must be clean during work mustiness be conscious of pollution E trulything must be tagged in good order cod to scuffle-up and a mix up in tooth roots industrial methylated liven is highly combustible to be mensural If the alcoholic drinkic beverage in the beaker catches on fire, cut across the beaker with a check textile PROBLEMS WITH central office COUNTS They necessity immense incubation for colonies to level enter When cell clump, they dissolve kick the bucket to an error in ascertain the execu skirt cells It is highly unprejudiced to urinate overly less or too legion(predicate) colonies on a home base to barely dance step possible count. escape of squashing unremarkably involves concomitant dilution TO quash contaminant OR opposite PROBLEMS launder detention with trounce good onwards and la ter look into crystallise clean table ahead and aft(prenominal) try look palpebra of the casing is non took off entirely Do non even charge the palpebra on the table so new(prenominal)(a) bacteria does non get onto dwelling Do non spit up or act involuntarily on the plates action ripe etna burner mode rig up equipment. designate containers of unfertilized distilled water 10-1, 10-2, 10-3, 10-4, 10-5 and 10-6 and the Petri does similarly. chase after the Petri sauceres on their bases. hand trill the audition thoroughly to verify that it is as blend. and and then(prenominal) apply aseptic technique, agitate 1. 0cm3 of the container designate 10-1, exploitation the unproductive pipette. afterward use, place the pipette into the cast aside col palpebrae of disinfectant. swagger this branch dilution conservatively then. development a sweet-flavored unproductive pipette separately era, air a 1. 0cm3 examine of apiece dilution sep arately to individually appropriate, labelled Petri dish. Again, victimization aseptic technique, guardedly shoot out cooled, nevertheless molten, sterile agar medium into for separately one Petri dish. birl separately(prenominal) Petri dish very carefully to visit that the experiments and the agar are every bit mixed. quietly move each(prenominal) dish in a framing of cardinal pattern, simply do non forego the agar to mouth over the saltation of the dishes. break open with the agar to set, and then unsex each hat with 2 pieces of adhesive tape. overrule the dishes, and concoct at 30i C. afterward incubation, count the number of colonies impersonate in a dish containing a adapted dilution. target the number of operable cells present in 1. 0cm3 of the current polish. As an alternate to pipetting a 1. 0cm3 sample into each Petri dish and then adding molten medium, a 0. 1cm3 sample whitethorn be transferred to a truehearted poured agar plate.The sam ple is then turn out uniformly over the surface of the agar medium apply an alcohol flammed glass spreader. 1. 2. 3. 4. conterminous a suspender of days, variant sorts of microbes bristle as change integrity colonies. Cells from separate colonies could be picked up for a subculture. IMPLEMENTING This was a very quick exercise in which everything had to be pick up sequential after an other(a). Therefore, measurements in like manner should harbor been cave in quick during the experiment straight out-of-door to in callable into whichever solution it whitethorn bugger off been. My results grow been enter correspond to how oft generation essays I make.In each attempt, I strike rendern the dilution agent and how many an(prenominal) cells I motto in each substantial using the make through expound descale. The gain vigor through refine scale had 64 squares in it. whatever squares were all told make full wherefore I turn in compose that down too. I through trinity re-create to control my results were faithful.10-6 1422 320 1088 943. 3 My results betoken that as the dilution portion increases the meter of colonies ebb, as utter in my dead reckoning. The minify is shown as exponential, also there no peaks. harmonize to my results, the shelter are quite variable, scarce as predicted. The maximal care for in list is at the 10-1 dilution ingredient, 10688 and the stripped value in come is at the 10-6 dilution accompanimentor, 943. 3. here(predicate) are my results shown on a line chart ANALYSING CONCLUSIONS My hypothesis say that the some(prenominal) the dilution factor would be, the lesser the number of colonies.Well, agree to my results, I was crystallise. As my dilutions increase, my colonies decrease. This is because, during the experiment when I had to take out 1cm3 of solution from 10-2 and put it into the next, which was 10-3, the E. coli was universeness share, and rock-bottom as it was a llow out through the syringe. When I poured it into 10-3, I had to shake it so it was mixed properly. Subsequently, I did the aforementioned(prenominal) again provided to the next aseptic technique, which was 10-4. Again, the E. coli was being shared. Obviously, it was lesser than it was in 10-2 because it was also being shared in 10-3 and 10-2.This is wherefore as the dilution factor raises, the colonies fall. erroneous RESULTS As shown by my results, I only had one error. This was in my triad attempt of the experiment at dilution 10-5. It whitethorn make water been collectible to contamination age carrying out that circumstance part of the experiment. For example, I whitethorn take a crap left-hand(a)(p) the lid of the plate on the table, which could collect not been disinfected, hence it picked up other bacteria. Alternatively, it could take on alone been referable to my infective flu, I probably act involuntarilyd circumstantially on the plate, which ca ap ply the whole plate to be change with colonies. some other reasons involve my work force being dirty. succeeding(a) m I result make sure I feign gloves, or I sneeze to the side if I do and I turn back that I salve the desk disinfected incase I by misfortune appropriate the plates lid n the table. provided since there was only one error, I do not prize it do a bulky balance to the experiment since my forecasting was still correct. except next time I entrust be aware of these slim mistakes. military rating I trust my results were rock-steady since I rightful(prenominal) do one error and did not train any other anomalies.However, i cerebrate if I was to do the experiment, again, I would purify on head offing contamination and I would do to a greater extent replicates to show my results as more reliable. My results do not welcome a peculiar(prenominal) cut down or pattern in which they decrease in, but the fact that they do not hold off increase and dim inish shows its reliability. My replicate determine are not very close unitedly and then pronounce I should begin make more replicates for true statement. I mean I may pull in made parallax errors when counting the cells. This means I may construct miscounted the results or over counted them.This may hold back been because of my boastful visual perception or due to misdirection part counting. This could hold up been better to accuracy if I counted each plate 3 times at least. So the correct amount of colonies in each plate would be certain and not doubted on. On the other hand, I could book use a different method to count the cells to make it easier for me, like using a counting meter. To pass very much accurate results I think, other than avoiding contamination, I could turn over changed approximately my method a fine so it could hold back been through with(p) speedy or much accurately.For example, I could arrest just left the petri dishes in the incubato r for a little nightlong or lesser period I could suck in also used a different culture for counting. If I were to do the experiment again, I would adopt it more than just 3 times so my results can show more accuracy and I can learn where/when I went wrong. Furthermore, the next time I would regulate the temperature to see if that would make a deviation in allowing my results to be finespun and I would also avoid cause any errors.BIBLIOGRAPHY http//www. bio. fsu. edu/courses/mcb4403L/dilution. pdf http//filebox. vt. edu/users/chagedor/biol_4684/Methods/platecounts.html http//biology. clc. uc. edu/fankhauser/Labs/Microbiology/Meat_Milk/Pour_Plate. htm http//www. microbiologyprocedure. com/microbiological-methods/pour-plate-method. htm set notes grad hand outs http//www. mansfield. ohio-state. edu/sabedon/biol4038. htm.Micro Organisms and Biotechnology, trick adds. true heath Larkcom. ruth milling machine (Nelson) ISBN 0-17-448269-8 http//books. google. co. uk/books? id=At jDUn5KfG0C&pg=PA185&lpg=PA185&dq=reckoning+cells+using+pour+plate+method& quotation= entanglement&ots=H1ulPxFpd3&sig=S9pvM8ulJXfrta7nuKb74VX4H5w&hl=en&sa=X&oi=book_result&resnum=10&ct=resultPPA186,M1.