Microbiology Lab: 2, 3, 4, 11

Ch2,3,411 CH#2 Brightfield micascopy bright field dark object can be used to obserre unstained microorganisms in wet mounts hanging drar slides false moverment Brownian movement the movement (hand strikes object Causes it to bounce vibrate at the same time, keep ther positions motile-achially more from one position to another movement w more directed loccassional spin/ros micmblames-microbial communities in environments Ex ratowa, algae, fung), bactena found in rond water. looking at Murrorganisms is useful in determining 31st, shape, and movement - wet mount useful, fast way to see bagena bus hard to see movement! Hanging dror techmque better for larger mcrobes and motility +(Petro kun jelly seal reduces evarirabon of the drop of flared better for * wet mount (the most basic ane)- STEPS (small/nonimonle suurpend infusion (Stirring/chaking) 2luse pausteor pirelse to transter a small diep onto a slide Handle coversip by its edges and place it of the drop gently press end of pencil or loop handle (to secure (5) Place on slyde on the meroscope and stars on lowes Port and go to the highest better for Hanging-Drop Procedure (bigger/motile) Dobtain dean hanging-dror (depression) slide Pick up a small amount of retroleum jelly on a toothpick of Pick UP a covriste (bells edges), and carefully touch the fetaleun jelly wlan edge the coverclir to get a sthal rim of Petroleum Jelly Repeat the other 3 edges, keeping the retroleum jelly on the same side d' the arsite @place bactene in middle of coverstip ⑤ place depression slide over the coverslip Quickly invest it dror should be hanging at this po view it under meeroscope Ist solid culture media was used in 1872 and mas made from Potato starch, coagulerted egg white, dod meat caseptic techninine art pimedures used to avoid introducing unwanted micnbes and to prescat spreading micmbes where they are wanted (huuleting loop Nichrome are held with on insulated hande Flaming-stenliung the inoculating tube before and after use (single-use plastes your are so framed-aiready sent Held over burner or in electine incenerator Joner be corner of blue lame (hottest part) gar comes from seg need -Angeling Hess suggested use of asar for growing bactenal cultures Microbiome community of meerobes that inhabit oa Parhuslar environment Environment microbiones con crist on lav benches environment objects? human microbemes on humanbody/Skin Most of time, are harmiss medioms bred to culture bacteno require macronutrents (carbon, niingin, Phosphorus, and sulfur + an energy source, and growin factors -chemically defined medium -a medium whose exact chemical compacitrons known is called a chemically defined medium which the exact chemical composition yanes slightly complex media: media for from batch to batch its complex) -purnent broth-commonly used ineud complex med nim Nutnent agar solidifzal nutrient both that had agar added to it Agar extract from manne red algae, Useful in culture media termains solid clunng mcrobial gronth lqueties @ 1000 remains lianid until is cooled at you Once solid, can be incubated at 100's and remain soind! -media needs sterilization after preparation most common steam sterilization @autoclaving heated to " steam undu pressunt. 121°C 91 15 Pounds of messure x 15 mins. growing bactena, petri plates contain solid media and alow @surtare area for examination Microbes are (noculated/ introduced to butrient agar(sund) or natment broth (liquid) Incubation recad-bacteno grome atter-bruth will become based /clandy=9muth -bacten ar gaith in broth determined Forbidity Colony agar-you will see bactena with naked elle growth pellicle on surface Hocesstent sospended in population of oplls that anses from a single bactenal cen sediment a CFO(Colony forming unit) colony that anses from a group of the bustorm same microbes attached to one another -preparing culture media prepare not of of mument broth in 250ml flask J 2 add louml of distilled water (3) add nutrient broth powder ✓ mix until the powder is dissolved✓ Pipette 5 ml of broth into each test tubes/ (3)and cap enc Label "Nutrient broth" (2 vwill be autoclaved, last one won't) ⑦ add agar to remaining 15 mi of notnent broth Boll until agar is dissolved ④ Label nutnent agar 2 (10) autoclare flask and previous tubes/ allow cooling at room temp of 45% water bolh pouning Plates faren front of you (unopened)✓ Petn/dishes (2) Hold fiask at angle remove stopper - 4th and 5th finger at other hand 4 Heat mouth of Hask 2-3 times 5 remove cover from first dish I sume hand thats holding storper (4) Pour agar Quickly/neatly (cover bottom about 5mp) replak cover others petridishes @Swirl all & plates to cover any empty spaces do touch lidandagar) Have slightly open 15 min to & condensat discard empty fiask in discard area Al culture medio are spentered prior to use tehihatan taccomplished using de autoclave Powaith cuttures provide laat of such a small quic and are cosity for Agar stems) tubes containing gold clture medle that weit ist at an angle and solida o qua quath surfare easy to store transport easier than petry plater) Agar deep agu sondhed at ibm of test tube end to edine thit gute lesso -semisolid agar only as 47 (Instead of usual17) 13 mottle bectens mots aney trys potenciation, conving apprennel of an imasted pine tree STAINS michime Angulla, endreportale diffemteater gramslam and acel fut star Inoculation which to use depends on th noculanna vor sed for and tentand meataring needle used for semisolid cry PS Mold inoculating toor in dominant bund broth tube mother hand edge of stenlite leer hottest nart (inner blue frame) (2-4. Stes) while holding sterk bor, remove cap of tube Hold it in hand Hold to be in front of stenliver 5 Immerse the sterne loup into broth cand get calline Oremove loor #resteriline mouth of fube. recaptove repeat -always hold culture taber and unnatulased wors at about a 20 ang p jhiniwire the amount of dust that cound fo and dous tis Moe too far blk the liony will leak out dovit yor of tobe hurd anything Powing batrena enhances the caritast between backend and the mounding Naren and vermys i laservations of analer deont and ressentir do my hunt Procedures gimate vain toy Focenent is ased and all betterna are similgily'stained (sume dye) s mathew closenes are od sed and beckena reactidifferentise to be Khurasalu viedt denty speeps of michoropropens ved cultures(more thans (mory/backng) haider 14 та ства пилоти Lasures & PURE Curve (comams | Minare In 1880 Rubettach prepared solid bredie that helped to slote M Audy more In depth Pure cultures Lisolating bactena &dilution methods are used Ostreak Plate-most-commonley used spread plate your plate Solid streak loor is used to streak mixed sample many times over culture medium in a Petri Disl -streaking repetendly causes bactena to fall off loop one by. and ore and be distributed over the agar surface, where each cell develops into a colony STEPS label bottom of Pen Dish. 2 Flame 100% attain loopful of broth Culture. etri cand liftone edge of petricorer, hold in same hand and du Ist streak One 5 Flame loor Y hold Your Tum Plate-Streakagan 7 Flame loop etc etc. -areas that touch are reineuntarted (gang back across aprenons scation) 2) Spread mate technvr ne-small amount of previously diluted specimen is spread over the surface of a solid medium using a spreading, rool. pour Plate technique-small amount of diluted sample. Is mixed I melted agar and poured into empty, sterile Petri Dishes. STEPS Dlabel bottom of sperridisher Lisolating bactena &dilution methods are used Ostreak Plate-most-commonley used spread plate your plate Solid streak loor is used to streak mixed sample many times over culture medium in a Petri Disl -streaking repetendly causes bactena to fall off loop one by. and ore and be distributed over the agar surface, where each cell develops into a colony STEPS label bottom of Pen Dish. 2 Flame 100% attain loopful of broth Culture. etri cand liftone edge of petricorer, hold in same hand and du Ist streak One 5 Flame loor Y hold Your Tum Plate-Streakagan 7 Flame loop etc etc. -areas that touch are reineuntarted (gang back across aprenons scation) 2) Spread mate technvr ne-small amount of previously diluted specimen is spread over the surface of a solid medium using a spreading, rool. pour Plate technique-small amount of diluted sample. Is mixed I melted agar and poured into empty, sterile Petri Dishes. STEPS Dlabel bottom of sperridisher Speual media ennched media Ex: Blood agar Chocolate agar medid thaver martin medium eosin methylene blue campylobacter agar Differenttal medig macconkey agar CLED agar TCBS agar Lowenstein-Jenson method. XLD agar Transport media Anaerobic media obligate gerobes Facultative anderobes Aerototerant angenobes