Cov txheej txheem staining thiab microscopic soj ntsuam ntawm cov kab mob
Yooj yim Staining ntawm cov kab mob
1 Lub Hom Phiaj
1.1 Kawm cov tswv yim yooj yim ntawm microbial smear thiab staining.
1.2 Yuav kom paub qhov yooj yim staining txoj kev ntawm cov kab mob.
1.3 Txhawm rau kom paub txog cov yam ntxwv ntawm cov kab mob morphological, thiab sib sau ua ke kev kawm ntawm kev siv roj iav thiab cov txheej txheem ua haujlwm aseptic.
2 Txoj Cai Kev smear 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, lawv tsis yooj yim rau kev paub nyob rau hauv lub teeb pom kev zoo tib yam, thiab lawv yuav tsum tau stained. Kev siv ib qho zas xim rau stain cov kab mob, kom cov kab mob stained thiab keeb kwm yav dhau los tsim cov xim sib txawv, kom lawv cov morphology thiab cov qauv tuaj yeem pom meej dua. Txoj kev no yog ib qho yooj yim rau kev ua thiab yog qhov tsim nyog rau kev soj ntsuam ntawm cov duab dav dav ntawm cov kab mob thiab kev teeb tsa ntawm cov kab mob. Alkaline dyes feem ntau yog siv rau staining yooj yim, 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 alkaline dyes yog ionized, staining feem ntawm lawv cov molecules yog zoo them, yog li staining feem ntawm alkaline dyes. tau yooj yim khi rau cov kab mob kom cov kab mob muaj xim. Cov kab mob stained cov kab mob sib txawv nrog cov keeb kwm yav dhau thiab yooj yim dua rau kev paub hauv lub tshuab tsom. Feem ntau siv dyes rau staining yooj yim muaj xws li merocyanine xiav, siv lead ua violet, thiab yooj yim compound liab. Thaum cov kab mob decomposition ntawm cov piam thaj los tsim cov kua qaub kom cov pH ntawm qhov nruab nrab txo qis, qhov zoo ntawm cov kab mob nce, thiab tom qab ntawd tuaj yeem siv cov xim liab, acidic liab lossis Congo liab thiab lwm yam acidic dyes staining. Cov kab mob yuav tsum tau kho ua ntej staining. Nws lub hom phiaj yog ob npaug: ib qho yog tua cov kab mob thiab ua kom cov kab mob ua rau cov swb; qhov thib ob yog kom nws affinity rau cov xim. Muaj ob txoj hauv kev uas feem ntau siv: cua sov thiab tshuaj kho. Sim ua kom tus thawj morphology ntawm cov hlwb thaum kho.
3 Cov khoom siv
3.1 Strain Bacillus subtilis 12-18h nutrient agar slant culture, Staphylococcus aureus txog 24h nutrient agar slant culture, Escherichia coli 24h nutrient agar slant culture, baker's poov agar slant culture.
3.2 Stain Lue's alkaline methylene xiav stain (los yog ammonium oxalate crystal violet stain), carbolic acid compound liab stain.
3.3 Cov cuab yeej siv los yog lwm yam khoom siv hauv Microscope, lub teeb dej cawv, swb, lub nplhaib inoculation, swb tuav, ob-txheej lub raj mis (sau nrog cedar roj thiab xylene), daim ntawv kab xev, lub cev ntsev los yog dej distilled, thiab lwm yam.
4 Cov txheej txheem Smear → ziab → fixation → staining → ntxuav → ziab → microscopic kuaj.
5 Cov kauj ruam
5.1 smear Siv ob lub qhov dej huv thiab tsis muaj roj, txhua qhov me me ntawm cov ntsev (los yog cov dej distilled) nyob rau hauv qhov chaw ntawm cov swb nyob rau hauv aseptic mob, nrog lub nplhaib inoculating rau aseptically khiav lag luam, raws li, los ntawm Bacillus subtilis, Garcinia micrococcus thiab Escherichia coli slant xaiv me ntsis moss hauv dej, sib tov thiab coated nrog ib zaj duab xis. Yog tias cov kab mob raug tshem tawm (lossis cov kab lis kev cai ua kua) smear, tuaj yeem siv los kho lub nplhaib xaiv 2 mus rau 3 lub nplhaib ncaj qha rau ntawm qhov swb. Nco ntsoov tias cov saline (dej distilled) thiab noj cov kab mob yuav tsum tsis txhob ntau dhau thiab kis tusyees, tsis tuab heev.
5.2 Kev ziab kom qhuav ib txwm nyob ntawm chav tsev kub. Nws tuaj yeem qhuav los ntawm cua sov me ntsis saum lub teeb cawv nrog rau sab coated. Txawm li cas los xij, tsis txhob nyob ze ntawm cov nplaim taws, vim tias qhov kub thiab txias heev yuav ua rau cov kab mob morphology puas.
5.3 Fixation Yog tias siv cov cua sov kom qhuav, kho thiab ziab yog muab tso rau hauv ib kauj ruam tib yam li ziab. Smear, qhuav thiab cua sov kho.
5.4 Staining Muab cov zawv zawg tiaj tus rau ntawm lub txee swb, ntxiv 1 mus rau 2 tee ntawm cov tshuaj pleev xim rau ntawm cov smear (cov tshuaj pleev xim tsuas yog npog cov smear zaj duab xis yog tsim nyog). Stain lub smear nrog Lü's Basic Mellanox Blue rau 1 ~ 2min, thiab nrog Carbonate Liab (los yog Ammonium Oxalate Crystal Violet) li 1min.
5.5 Rinsing Ncuav cov tshuaj staining 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 yaug, tsis txhob cia cov dej ntws ncaj qha los ntxuav cov coated nto. Cov dej ntws yuav tsum tsis txhob nrawm dhau los yog loj heev kom tsis txhob cuam tshuam cov smear zaj duab xis.
5.6 Drying co tawm cov tee dej ntawm tus swb kom qhuav ib txwm, tshuab ziab lossis nqus cov ntawv tuaj yeem siv los nqus kom qhuav (ceev faj tsis txhob so cov kab mob hauv lub cev). 5.7 Kev soj ntsuam Microscopic Lub smear yog qhuav thiab tshuaj xyuas los ntawm microscopy. Cov smear yuav tsum tau qhuav tag ua ntej nws tuaj yeem pom nrog lub tshuab ntsuas roj.
6 Cov txiaj ntsig Kos lub morphology ntawm E. coli thiab Micrococcus garciniae tom qab ib qho staining.
Gram Stain
1 Lub Hom Phiaj
1.1 Kom nkag siab lub hauv paus ntsiab lus ntawm Gram stain thiab nws qhov tseem ceeb hauv kev faib tawm thiab kev txheeb xyuas cov kab mob.
1.2 Kawm thiab paub cov txheej txheem Gram staining thiab sib sau ua ke kev kawm ntawm kev siv roj lo ntsiab muag ntawm lub teeb tsom iav.
