Cov txheej txheem staining thiab kev soj ntsuam microscopic ntawm microorganisms
1. Lub hom phiaj
1.1 Kawm cov txheej txheem hauv paus ntawm microbial smears thiab staining.
1.2 Tus tswv txoj kev staining yooj yim ntawm cov kab mob.
1.3 Ua ntej nkag siab txog cov yam ntxwv morphological ntawm cov kab mob, thiab sib sau ua ke kev paub txog kev siv roj lo ntsiab muag thiab cov txheej txheem ua haujlwm aseptic.
2 Cov ntsiab cai
Smearing thiab staining ntawm cov kab mob yog cov txheej txheem yooj yim hauv kev sim microbiology. Cov kab mob ntawm cov kab mob me me thiab pob tshab, ua rau lawv nyuaj rau txheeb xyuas raws li lub teeb pom kev zoo tib yam; lawv yuav tsum tau stains. Ib qho zas xim yog siv los zas cov kab mob, kom cov kab mob stained tsim cov xim sib txawv ntawm keeb kwm yav dhau, kom lawv cov morphology thiab cov qauv tuaj yeem pom meej dua. Txoj kev no yog ib qho yooj yim rau kev khiav lag luam thiab tsim nyog rau kev soj ntsuam cov qauv ntawm cov kab mob thiab cov kab mob. Basic dyes feem ntau yog siv rau staining yooj yim. Qhov no yog vim hais tias nyob rau hauv nruab nrab, alkaline los yog tsis muaj zog acidic daws, cov kab mob hlwb feem ntau tsis zoo nqi, thiab thaum cov dyes yooj yim ionized, dyeing ib feem ntawm lawv cov molecules yog zoo them, yog li ntawd lawv yog alkaline. Cov staining ib feem ntawm cov dye tau yooj yim ua ke nrog cov kab mob kom xim rau lawv. Cov kab mob stained cov kab mob sib txawv nrog cov keeb kwm yav dhau, ua rau lawv yooj yim dua los txheeb xyuas hauv lub tshuab tsom. Cov dyes feem ntau siv rau kev dyeing yooj yim xws li methylene xiav, siv lead ua violet, yooj yim fuchsin, thiab lwm yam. Thaum cov kab mob decompose qab zib thiab tsim kua qaub, ua rau pH ntawm kab lis kev cai nruab nrab poob, tus nqi zoo ntawm cov kab mob nce. Lub sijhawm no, cov kua qaub dyes xws li eosin, acid fuchsin, lossis Congo liab tuaj yeem siv rau staining. Cov kab mob yuav tsum tau kho ua ntej staining. Nws muaj ob lub hom phiaj: ib qho yog tua cov kab mob thiab ua kom lawv ua raws li cov iav swb; lwm qhov yog kom lawv cov affinity rau dyes. Ob txoj kev siv feem ntau yog cua sov thiab tshuaj kho. Thaum kho, sim ua kom zoo li qub ntawm cov hlwb.
3 cov ntaub ntawv
3.1 Bacteria: Bacillus subtilis 12-18h nutrient agar slant culture, Staphylococcus aureus 24h nutrient agar slant culture, Escherichia coli 24h nutrient agar slant culture, baker's poov agar slant culture.
3.2 Dyeing agent: Lu's alkaline methylene xiav dye (los yog ammonium oxalate crystal violet dye), carbolic acid fuchsin dye.
3.3 Cov cuab yeej siv los yog lwm yam khoom siv: lub tshuab tsom iav, lub teeb dej cawv, swb, inoculation voj, swb khib, ob-txheej lub raj mis (muaj cov roj cedar thiab xylene), cov ntaub so ntswg ntawm lub cev, saline los yog dej distilled, thiab lwm yam.
4. txheej txheem: smear → ziab → fixation → staining → ntxuav → ziab → microscopic kuaj.
5 qib
5.1 Smear: Siv ob lub iav huv thiab tsis muaj roj. Nyob rau hauv cov xwm txheej tsis muaj menyuam, muab ib qho me me ntawm cov ntsev ib txwm (los yog dej distilled) rau ntawm txhua qhov chaw ntawm lub khob swb. Siv lub voj inoculation los ua kom tsis muaj kab mob los ntawm Bacillus subtilis. , Micrococcus luteus thiab Escherichia coli, khaws cov kab mob me me hauv cov dej ntws ntawm cov slant, sib tov zoo thiab siv rau hauv cov yeeb yaj kiab nyias. Yog tias koj siv cov kab mob tua kab mob (los yog kua kab lis kev cai) smear, koj tuaj yeem siv lub voj inoculating los xaiv 2 mus rau 3 lub voj voog thiab siv ncaj qha rau ntawm qhov swb. Ceev faj kom tsis txhob tso dej ntau dhau ntawm lub cev (cov dej distilled) thiab tshem tawm cov kab mob thiab siv nws sib npaug thiab tsis tuab heev.
5.2 Kev ziab kom qhuav ib txwm nyob ntawm chav tsev kub. Koj tuaj yeem ua kom sov nws me ntsis dhau lub teeb cawv nrog rau sab coated tig mus kom qhuav. Tab sis nco ntsoov tsis txhob mus ze ntawm cov nplaim taws, vim qhov kub thiab txias yuav ua rau cov kab mob morphology puas.
5.3 Fixation Yog siv cov cua sov kom qhuav, kho thiab ziab kom qhuav yog ua ke rau hauv ib kauj ruam, thiab txoj kev yog tib yam li ziab. Smear, qhuav thiab cua sov-kho.
5.4 Staining: Muab cov swb tiaj tus rau ntawm qhov swb, thiab ntxiv 1 mus rau 2 tee ntawm cov tshuaj zas xim rau cov smear (nws yog qhov tsim nyog rau cov tshuaj zas zas tsuas yog npog cov smear zaj duab xis xwb). Stain nrog Lu's yooj yim methylene xiav rau 1 mus rau 2 feeb, thiab stain nrog carbolic acid fuchsin (los yog ammonium oxalate crystal violet) li ntawm 1 feeb.
5.5 Ntxuav nrog dej: Ncuav cov tshuaj zas xim thiab maj mam yaug nrog dej kais dej los ntawm ib qho kawg ntawm tus swb kom txog thaum cov dej ntws los ntawm cov smear tsis muaj xim. Thaum ntxuav nrog dej, tsis txhob ntxuav lub coated nto ncaj qha nrog dej. Cov dej ntws yuav tsum tsis txhob nrawm dhau los yog loj dhau los tiv thaiv cov smear zaj duab xis los ntawm kev poob.
5.6 Qhuav: Muab cov tee dej ntawm cov swb thiab qhuav ib txwm, tshuab ziab plaub hau los yog nqus nrog cov ntawv nqus dej (ceev faj tsis txhob so cov kab mob). 5.7 Kev tshuaj xyuas microscopic Tom qab cov smear qhuav lawm, ua cov tshuaj microscopic. Cov smear yuav tsum tau qhuav tag ua ntej saib nrog lub lens roj.
6 Cov txiaj ntsig Kos lub morphology ntawm E. coli thiab Micrococcus luteus pom tom qab ib qho staining.
