Dab tsi ntawm microscope yog siv los saib cov duab ntawm cov kab mob microbial
Ib lo lus sib sau ua ke rau tag nrho cov kab mob me me uas nyuaj rau tib neeg los saib ntawm qhov muag liab qab. Cov kab mob muaj xws li kab mob, kab mob, fungi, thiab ob peb algae. (Txawm li cas los xij, qee cov kab mob tuaj yeem pom ntawm qhov muag liab qab, xws li nceb uas muaj cov kab mob fungi, Ganoderma lucidum, thiab lwm yam) Cov kab mob yog ib hom "cov kab mob uas tsis yog cellular" uas muaj ob peb yam xws li nucleic acids thiab proteins, tab sis lawv kev ciaj sia yuav tsum nyob ntawm cov hlwb muaj sia. Raws li cov chaw sib txawv uas muaj nyob, lawv tuaj yeem muab faib ua cov kab mob prokaryotic, cov kab mob hauv qhov chaw, cov kab mob fungal, cov kab mob poov xab, cov kab mob marine, thiab lwm yam.
Lub luag haujlwm thiab kev puas tsuaj ntawm microorganisms:
Ib qho kev cuam tshuam tseem ceeb tshaj plaws ntawm cov kab mob microorganisms rau tib neeg yog cov kab mob sib kis. 50 feem pua ntawm tib neeg cov kab mob yog tshwm sim los ntawm cov kab mob. Cov keeb kwm ntawm microbes ua rau tib neeg muaj kab mob yog keeb kwm ntawm tib neeg kev tawm tsam tsis tu ncua nrog lawv. Tib neeg tau ua tiav zoo hauv kev tiv thaiv thiab kho cov kab mob, tab sis cov kab mob tshiab thiab rov tshwm sim ntawm cov kab mob microbial txuas ntxiv mus, xws li cov kab mob loj heev uas tsis muaj cov tshuaj kho tau zoo. Cov txheej txheem pathogenic ntawm qee cov kab mob tsis meej. Kev tsim txom ntawm ntau cov tshuaj tua kab mob dav dav tau ua rau muaj kev xaiv muaj zog, ua rau ntau hom kev hloov pauv, ua rau muaj qhov tshwm sim ntawm cov tshuaj tiv thaiv, thiab tib neeg kev noj qab haus huv raug hem los ntawm kev hem thawj tshiab. Qee cov kab mob segmented tuaj yeem hloov pauv los ntawm kev rov ua dua lossis rov ua dua tshiab. Qhov piv txwv tshaj plaws yog tus kab mob khaub thuas.
Tom qab paub txog cov ntsiab lus tshwj xeeb ntawm cov kab mob, cov kab mob dab tsi yuav tsum tau siv thaum kawm cov kab mob kom pom, thiab siv lub tshuab ntsuas qhov twg tuaj yeem pom zoo dua, thiab soj ntsuam thiab txheeb xyuas cov kab mob sib kis.
Lub invention ntawm lub microscope yog kom pom cov khoom luag ntxhi uas tsis tuaj yeem pom ntawm qhov muag liab qab. Qhov loj ntawm cov kab mob me me heev, yog li lawv yuav tsum tau nthuav dav thiab saib xyuas nrog kev pab los ntawm lub tshuab tsom. Tsis tas li ntawd, muaj ntau hom kab mob, yog li feem ntau ntawm cov kab mob optical microscopes tuaj yeem soj ntsuam cov kab mob, cov lus nug tom ntej yog hom kab mob twg yuav tsum tau siv rau kev soj ntsuam thiab tshuaj xyuas cov kab mob. Cov tshuab microscopes feem ntau rau kev soj ntsuam microbial morphology suav nrog cov kab mob lom neeg, cov kab mob sib txawv, cov kab mob sib txawv, cov kab mob fluorescence microscopes, thiab confocal microscopes. Microscope thiab lwm yam.
Cov hauv qab no piav qhia txog ntau yam microscopes siv los soj ntsuam cov kab mob:
1. Ordinary light microscope
Lub teeb ntuj lossis lub teeb yog siv los ua lub teeb ci, thiab nws lub wavelength yog li ntawm {{0}}.4 μm. Qhov kev daws teeb meem ntawm lub tshuab microscope yog ib nrab ntawm lub wavelength, uas yog, 0.2 μm, thiab cov duab me tshaj plaws pom ntawm qhov muag liab qab yog 0.2 hli. Yog li ntawd, siv cov roj (immersion) daim iav los nthuav 1000 zaug tuaj yeem ua kom loj dua 0.2μm cov khoom rau hauv 0.2mm pom ntawm qhov muag liab qab. Ordinary optical microscopes tuaj yeem siv rau kev soj ntsuam cov kab mob, actinomycetes thiab fungi.
2. Darkfield microscopy feem ntau yog siv los soj ntsuam cov kab mob microbial tsis muaj kab mob thiab kev txav mus los. Tom qab qhov tsaus nti condenser tau teeb tsa rau hauv lub tshuab tsom iav zoo tib yam, lub teeb tsis tuaj yeem nkag ncaj qha los ntawm nruab nrab, thiab qhov pom ntawm qhov tsaus. Thaum cov qauv tau txais oblique lub teeb los ntawm ntug ntawm lub condenser, nws tuaj yeem tawg, yog li cov kab mob ci ntsa iab tuaj yeem pom hauv thaj chaw tsaus ntuj xws li kab mob lossis spirochetes.
3. Phase contrast microscope Phase contrast microscope siv lub teeb nyhuv ntawm theem sib txawv phaj hloov lub teeb theem thiab amplitude ntawm lub teeb ncaj qha, thiab hloov qhov sib txawv ntawm lub teeb theem mus rau lub teeb sib txawv. Nyob rau hauv ib lub sij hawm-contrast microscope, thaum lub teeb dhau los ntawm cov qauv tsis muaj xim, qhov sib txawv ntawm lub teeb theem yog tshwm sim los ntawm qhov tsis sib xws ntawm qhov ntom ntawm qhov sib txawv ntawm cov qauv, thiab cov morphology, cov qauv sab hauv thiab kev txav ntawm cov kab mob tuaj yeem pom.
4. Fluorescence microscope Fluorescence microscope yog qhov pib zoo ib yam li cov tshuab kho qhov muag zoo tib yam, qhov sib txawv tseem ceeb yog lub teeb ci, lim thiab condenser. Tam sim no, feem ntau ntawm lawv siv cov khoom siv hluav taws xob, thiab cov teeb hluav taws xob siab mercury feem ntau yog siv los ua lub teeb pom kev zoo, uas tuaj yeem tso tawm ultraviolet lossis xiav-violet teeb. Muaj ob hom lim: excitation lim thiab nqus lim. Ntxiv rau cov condensers dav dav dav dav, qhov tsaus nti-teeb condensers tuaj yeem siv rau hauv fluorescence microscopes siv lub teeb xiav los txhim kho qhov sib txawv ntawm fluorescence thiab keeb kwm yav dhau. Txoj kev no siv tau rau kev tshawb nrhiav lossis kev txheeb xyuas cov kab mob uas muaj cov xim fluorescent lossis ua ke nrog cov tshuaj tiv thaiv fluorescent.
5. Electron microscopes siv electron flow raws li lub teeb ci. Piv nrog rau lub teeb pom kev, lub wavelength yog kaum tawm txhiab lub sij hawm sib txawv, uas zoo heev txhim kho qhov kev daws teeb meem. Lub tshuab nqus hlau nplaum yog siv los ua qhov optical amplification system, thiab qhov magnification tuaj yeem ncav cuag kaum txhiab lossis ntau pua txhiab zaus. Nws feem ntau yog siv rau hauv cov kab mob. thiab kev soj ntsuam ntawm cov kab mob ultrastructure.
Kev soj ntsuam ntawm unstained microbial specimens:
Cov qauv tsis muaj xim feem ntau tuaj yeem siv los soj ntsuam cov kab mob morphology, lub zog thiab kev txav. Cov kab mob tsis muaj xim thiab pob tshab thaum tsis muaj xim, thiab tau pom nyob rau hauv lub tshuab tsom iav feem ntau los ntawm qhov sib txawv ntawm qhov ntsuas qhov ntsuas ntawm cov kab mob thiab ib puag ncig ib puag ncig. Cov kab mob nrog flagella txav mus rau lub zog, thaum cov kab mob uas tsis muaj flagella qhia qhov tsis xwm yeem Brownian motion. Cov kab mob zoo xws li Treponema pallidum, Leptospira, thiab Campylobacter muaj cov duab sib txawv thiab cov qauv txav, uas yog qhov tseem ceeb rau kev kuaj mob. Feem ntau siv txoj kev yog txoj kev poob siab, pendant drop method thiab capillary method.
1. Txoj kev dai dai Siv vaseline nyob ib ncig ntawm lub qhov concave ntawm lub qhov huv si concave iav swb, nqa lub nplhaib ntawm cov kab mob tshem tawm nrog ib lub voj inoculation thiab muab tso rau hauv nruab nrab ntawm lub hau npog iav, tom qab ntawd kho qhov concave qhov ntawm lub iav concave nrog. lub tee dej nyob rau hauv nruab nrab ntawm lub khob iav thiab muab lub hau rau, ces tig nws dua sai sai, maj mam nias lub hau ntswj kom nws lo nruj nreem rau lub vaseline ntawm ntug ntawm lub concave qhov, thiab ces saib nyob rau hauv ib tug high-power. microscope (los yog qhov tsaus ntuj).
2. Siv lub nplhaib ntawm cov kab mob tshem tawm nrog ib lub voj inoculation thiab muab tso rau hauv qhov chaw ntawm lub iav huv si los ntawm txoj kev poob siab, thiab maj mam npog cov kab mob tshem tawm nrog lub hau npog iav, saib xyuas kom tsis txhob muaj cua npuas thiab tiv thaiv. cov kab mob raug tshem tawm los ntawm overflowing. Brightfield (los yog darkfield) kev soj ntsuam nyob rau hauv lub siab lub hwj chim lens.
3. Txoj kev capillary feem ntau yog siv rau kev kuaj xyuas cov kinetics ntawm cov kab mob anaerobic. Feem ntau xaiv 60~70mm ntev. Tom qab siphoning cov kab mob anaerobic raug tshem tawm los ntawm lub capillary nrog lub aperture ntawm 0. Lub capillary tau tsau rau ntawm lub iav swb nrog cov ntawv yas, thiab pom nyob rau hauv ib lub hwj chim loj hauv qhov tsaus ntuj.
Kev soj ntsuam ntawm stained microbial specimens nrog lub tshuab tsom:
Tom qab cov kab mob stained, vim qhov sib txawv ntawm cov xim ntawm cov kab mob thiab cov cheeb tsam ib puag ncig, cov yam ntxwv ntawm cov kab mob (xws li qhov loj, cov duab, kev teeb tsa, thiab lwm yam) ntawm cov kab mob thiab qee cov qauv tshwj xeeb tuaj yeem ua tau. pom meej meej nyob rau hauv ib qho yooj yim optical microscope (xws li tshuaj ntsiav, flagella, spores, thiab lwm yam), thiab cov kab mob tuaj yeem raug cais tawm thiab txheeb xyuas raws li cov tshuaj tiv thaiv staining.
(1) Cov txheej txheem dav dav ntawm cov kab mob staining Cov txheej txheem dav dav ntawm cov kab mob staining yog: smear (drying)-fixation-staining.
1. Smear Kev npaj cov ntshav, tso zis, tso tawm, puncture kua thiab kua kab lis kev cai, thiab ncaj qha nyias zaj duab xis smears rau iav slides; autopsy los yog kab mob cov ntaub so ntswg, smear qhov txhab nrog ib tug paj rwb swab rau sampling. Rau kev npaj cov kab mob kab mob los yog cov nyom ntawm cov khoom nruab nrab, ua ntej siv lub voj inoculation coj ib lub nplhaib ntawm saline ib txwm thiab muab tso rau hauv nruab nrab ntawm cov swb, tom qab ntawd siv lub voj tsis muaj kab mob kom coj me me ntawm kab lis kev cai thiab zom nws. tusyees nyob rau hauv ib txwm saline, thiab kis mus rau 1cm2 loj los yog me me pleev xim rau qhov chaw, cia nws qhuav nyob rau hauv chav tsev kub los yog qhuav maj mam nyob rau hauv ib tug deb.
2. Lub hom phiaj ntawm fixation yog tua cov kab mob, coagulate cov kab mob protein thiab cov qauv, thiab pab staining; txhawb cov kab mob kom ua raws li cov swb kom tsis txhob raug ntxuav los ntawm dej thaum ntxuav; hloov cov permeability ntawm cov kab mob mus rau dyes, uas yog zoo rau cov qauv ntawm cov kab mob hlwb ntawm staining. Nws feem ntau yog kho los ntawm cua sov nrog cov nplaim taws, thiab cov smear qhuav yog sai dhau los ntawm cov nplaim taws rau 3 zaug. Nws yog qhov zoo dua tsis txhob hlawv cov tawv nqaij ntawm sab nraub qaum thaum nws kov cov swb.
3. Dyeing Raws li kev soj ntsuam sib txawv, xaiv cov txheej txheem dyeing sib txawv rau dyeing. Thaum dyeing, ntxiv cov tshuaj dye dropwise kom qhov kev pab cuam.
4. Mordant Txhua yam khoom uas tuaj yeem txhim kho kev sib raug zoo ntawm cov xim thiab cov khoom dyed, kho cov xim ntawm cov khoom dyed thiab ua rau muaj kev hloov pauv hauv permeability ntawm cov cell membrane hu ua mordant. Feem ntau siv yog alum, tannic acid, hlau ntsev thiab iodine, thiab lwm yam, thiab cua sov kuj yog siv los txhawb cov xim. Mordants tuaj yeem siv nruab nrab ntawm thawj qhov staining thiab counterstaining, thiab tseem tuaj yeem siv tom qab fixation los yog muaj nyob rau hauv fixative thiab staining.
5. Decolorization Txhua yam tshuaj uas tuaj yeem tshem tawm cov xim ntawm cov khoom dyed yog hu ua decolorizer. Ethanol, acetone, thiab lwm yam yog siv los ua decolorizers. Tus neeg saib xyuas decolorizing tuaj yeem kuaj xyuas qhov kev ruaj ntseg ntawm kev sib xyaw ntawm cov kab mob thiab cov dyes, uas tuaj yeem siv rau cov xim sib txawv.
6. Cov tshuaj tiv thaiv kab mob los yog lawv cov qauv uas tau decolorized feem ntau counterstained nrog cov tshuaj counterstain kom yooj yim saib. Cov xim ntawm cov tshuaj counterstaining txawv los ntawm cov thawj dyeing tov los tsim ib tug ntse sib piv. Lub counterstaining yuav tsum tsis txhob muaj zog heev, thiaj li tsis mus npog cov xim ntawm thawj staining.
