Kev nqus thiab rov ua dua tshiab ntawm lub teeb los ntawm inorganic thiab organic media yog qhov tshwm sim ntawm phosphorescence lossis fluorescence. Qhov sib txawv ntawm qhov tshwm sim yog qhov ntev ntawm lub caij nyoog ntawm lub teeb nqus thiab emission ntawm cov kwj deg. Nrog fluorescence, cov txheej txheem no tshwm sim yuav luag ib txhij, thiab nrog phosphorescence, nrog rau qee qhov qeeb.
keeb kwm keeb kwm
Hauv xyoo 1852, tus kws tshawb fawb Askiv Stokes thawj zaug piav txog fluorescence. Nws tau sau lo lus tshiab los ntawm nws qhov kev sim nrog fluorspar, uas tawm lub teeb liab thaum raug ultraviolet teeb. Stokes tau sau tseg qhov tshwm sim nthuav. Nws pom tias lub wavelength ntawm lub teeb fluorescent yog ib txwm ntev dua li lub teeb excitation.
Ntau qhov kev sim tau ua nyob rau xyoo 19th kom paub meej qhov kev xav. Lawv pom tias muaj ntau yam qauv fluoresce thaum raug ultraviolet teeb. Cov ntaub ntawv suav nrog, ntawm lwm tus, muaju, resins, minerals, chlorophyll,tshuaj raw khoom, inorganic compounds, vitamins, roj. Kev siv ncaj qha ntawm cov dyes rau kev tshuaj xyuas lom neeg pib tsuas yog xyoo 1930
Fluorescence microscopy piav qhia
Qee yam ntawm cov ntaub ntawv siv hauv kev tshawb fawb hauv thawj ib nrab ntawm lub xyoo pua 20th yog qhov tshwj xeeb heev. Ua tsaug rau cov kev ntsuas uas tsis tuaj yeem ua tiav los ntawm txoj kev sib txawv, txoj kev fluorescence microscopy tau dhau los ua ib qho tseem ceeb hauv kev tshawb fawb txog kev kho mob thiab kev tshawb fawb lom neeg. Cov txiaj ntsig tau txais tsis tseem ceeb me me rau cov ntaub ntawv tshawb fawb.
Cov txiaj ntsig ntawm fluorescence microscopy yog dab tsi? Nrog kev pab los ntawm cov ntaub ntawv tshiab, nws tau dhau los ua kom cais cov hlwb tshwj xeeb thiab cov khoom siv submicroscopic. Lub fluorescent microscope tso cai rau koj los txheeb xyuas cov molecules ntawm tus kheej. Ntau hom dyes tso cai rau koj txheeb xyuas ntau lub ntsiab lus tib lub sijhawm. Txawm hais tias qhov kev daws teeb meem spatial ntawm cov cuab yeej raug txwv los ntawm kev txwv qhov sib txawv, uas, dhau los, nyob ntawm cov khoom tshwj xeeb ntawm cov qauv, kev tshawb pom ntawm cov molecules hauv qab no kuj tseem ua tau. Ntau cov qauv qhia autofluorescence tom qab irradiation. Qhov tshwm sim no tau siv dav hauv petrology, botany, semiconductor kev lag luam.
Ntse
Kev kawm txog tsiaj cov ntaub so ntswg los yog cov kab mob pathogenic feem ntau nyuaj los ntawm cov tsis muaj zog lossis muaj zog heev uas tsis yog tshwj xeeb autofluorescence. Txawm li cas los xij, tus nqi hauvkev tshawb fawb tau txais kev taw qhia rau hauv cov khoom ntawm cov khoom zoo siab ntawm ib lub wavelength tshwj xeeb thiab emitting lub teeb flux ntawm qhov yuav tsum tau siv. Fluorochromes ua raws li cov xim muaj peev xwm ntawm tus kheej txuas rau cov qauv (tsis pom lossis pom). Tib lub sijhawm, lawv txawv los ntawm kev xaiv siab nrog kev hwm rau lub hom phiaj thiab quantum yield.
Fluorescence microscopy tau dhau los siv dav nrog qhov tshwm sim ntawm ntuj thiab hluavtaws dyes. Lawv muaj qhov tshwj xeeb emissions thiab excitation siv profile thiab tau tsom rau cov hom phiaj lom neeg.
Kev txheeb xyuas ntawm tus kheej molecules
Ntau zaus, nyob rau hauv cov xwm txheej zoo, koj tuaj yeem sau npe lub ci ntawm ib lub caij. Txhawm rau ua qhov no, ntawm lwm yam, nws yog ib qho tsim nyog los xyuas kom meej txaus lub suab qis thiab kho qhov muag tom qab. Lub fluorescein molecule tuaj yeem tawm mus txog 300,000 photons ua ntej kev puas tsuaj vim photobleaching. Nrog rau 20% sau tus nqi thiab cov txheej txheem kev ua tau zoo, lawv tuaj yeem sau npe nyob rau hauv tus nqi ntawm 60 txhiab
Fluorescence microscopy, raws li avalanche photodiodes lossis electron sib npaug, tso cai rau cov kws tshawb fawb los soj ntsuam tus cwj pwm ntawm tus kheej molecules rau vib nas this, thiab qee zaum feeb.
Kev nyuaj
Qhov teeb meem tseem ceeb yog kev tawm suab nrov los ntawm kev kho qhov muag tom qab. Vim tias muaj ntau cov ntaub ntawv siv hauv kev tsim cov lim dej thiab lo ntsiab muag pom qee qhov autofluorescence, kev siv zog ntawm cov kws tshawb fawb hauv cov theem pib tau tsom mus rau kev tshaj tawm.cov khoom uas muaj fluorescence tsawg. Txawm li cas los xij, cov kev sim tom ntej tau ua rau cov lus xaus tshiab. Tshwj xeeb, fluorescence microscopy raws li tag nrho cov kev xav sab hauv tau pom kom ua tiav cov keeb kwm yav dhau los thiab cov teeb pom kev zoo siab.
Mechanism
Cov ntsiab lus ntawm fluorescence microscopy raws li tag nrho cov kev xav hauv sab hauv yog siv cov nthwv dej sai sai lossis tsis-propagating yoj. Nws tshwm sim ntawm qhov sib cuam tshuam ntawm cov xov xwm nrog cov ntsuas ntsuas sib txawv. Nyob rau hauv cov ntaub ntawv no, lub teeb beam dhau los ntawm ib tug prism. Nws muaj qhov ntsuas qhov ntsuas siab siab.
Lub prism yog nyob ib sab ntawm cov tshuaj aqueous lossis iav tsis muaj qhov ntsuas. Yog hais tias lub beam ntawm lub teeb yog qhia rau nws ntawm lub kaum sab xis uas siab tshaj qhov tseem ceeb, lub beam yog kiag li reflected los ntawm lub interface. Qhov no tshwm sim, nyob rau hauv lem, ua rau ib tug nonpropagating nthwv dej. Nyob rau hauv lwm yam lus, ib tug electromagnetic teb yog generated uas nkag mus rau ib tug nruab nrab nrog ib tug qis refractive index ntawm qhov deb ntawm tsawg tshaj li 200 nanometers.
Nyob rau hauv lub nthwv dej tsis-propagating, lub teeb siv zog yuav txaus txaus rau excite fluorophores. Txawm li cas los xij, vim nws qhov tob tob tob, nws qhov ntim yuav tsawg heev. Qhov tshwm sim yog keeb kwm qis qis.
Kev hloov kho
Fluorescence microscopy raws li tag nrho cov kev xav sab hauv tuaj yeem pom tau nrog epi-illumination. Qhov no yuav tsum tau lo ntsiab muag nrog nce tus lej aperture (tsawg kawg yog 1.4, tab sis nws yog qhov xav tau tias nws ncav cuag 1.45-1.6), nrog rau ib feem ntawm qhov pom ntawm lub cuab yeej. Cov tom kawg yog tiav nrog ib qho chaw me me. Txhawm rau kom muaj kev sib haum xeeb ntau dua, lub nplhaib nyias yog siv, los ntawm qhov twg ntawm qhov ntws tau thaiv. Txhawm rau kom tau txais lub kaum ntse ntse tom qab qhov kev xav tau tag nrho tshwm sim, qhov siab ntawm qhov cuam tshuam ntawm qhov nruab nrab immersion nyob rau hauv lub lo ntsiab muag thiab lub tsom iav tsom iav yog xav tau.
