Lub hauv paus ntsiab lus thiab cov yam ntxwv ntawm Fluorescence Microscopes
Lub teeb fluorescence feem ntau siv lub teeb hluav taws xob siab siab mercury (50-200 W), uas tawm lub teeb ntawm ntau lub wavelengths. Txawm li cas los xij, txhua yam khoom siv fluorescent muaj qhov tshwj xeeb excitation wavelength uas tsim cov fluorescence khaus. Vim li no, lub lim dej excitation (feem ntau muaj nyob rau hauv ultraviolet, violet, xiav thiab ntsuab hom) yuav tsum tso cai tsuas yog lub teeb excitation ntawm ib lub wavelength tshwj xeeb kom dhau mus thiab irradiate cov qauv, thaum nqus tag nrho lwm cov wavelengths.
Thaum irradiated los ntawm excitation light, txhua yam khoom emits pom fluorescence nrog ib tug ntev wavelength tshaj lub excitation teeb nyob rau hauv ib tug luv luv lub sij hawm. Fluorescence yog qhov tshwj xeeb thiab feem ntau tsis muaj zog dua li lub teeb pom kev zoo. Txhawm rau soj ntsuam qhov tshwj xeeb fluorescence no, yuav tsum tau teeb tsa lub teeb meem (lossis kev txwv) lim tom qab lub hom phiaj lens. Nws ua haujlwm rau ob lub hom phiaj: ua ntej, nws nqus thiab thaiv lub teeb pom kev zoo los ntawm kev nkag mus rau lub qhov muag kom tsis txhob cuam tshuam nrog kev soj ntsuam fluorescence thiab ua rau lub qhov muag puas; Thib ob, nws xaiv xa cov fluorescence tshwj xeeb los tsim cov xim fluorescent txawv. Ob hom lim yuav tsum raug xaiv thiab siv rau hauv kev sib tw ua khub.
Fluorescence microscopes muab faib ua ob hom raws li lawv txoj kev kho qhov muag:
1. Transmitted-light Fluorescence Microscope
Hauv qhov tsim no, lub teeb ci ntsa iab dhau los ntawm lub condenser thiab cov khoom siv coj los ua kom muaj zog fluorescence. Ib qho tsaus nti condenser feem ntau siv, txawm tias tus qauv condenser tuaj yeem siv tau; daim iav reflecting yog kho rau redirect thiab sab-irradiate lub excitation lub teeb mus rau cov qauv. Qhov no yog hom qub ntawm fluorescence microscope. Nws qhov zoo dua yog qhov muaj zog fluorescence ntawm qhov kev nthuav dav tsawg, thaum nws qhov tsis zoo yog qhov kev siv fluorescence txo qis thaum qhov kev nthuav dav nce. Yog li nws zoo dua rau kev soj ntsuam cov ntaub ntawv loj dua.
2. Epifluorescence Microscope
Qhov no yog ib qho niaj hnub, tsim tshiab hom fluorescence microscope. Tsis zoo li cov qauv tsim hluav taws xob, lub teeb pom kev zoo yog npaj rau hauv qab los ntawm lub hom phiaj lens mus rau qhov chaw kuaj - tib lub hom phiaj lens ua haujlwm ob qho tib si raws li lub teeb pom kev zoo thiab lub hom phiaj fluorescence-collecting. Lub dichroic beamsplitter yog ntsia rau hauv txoj kev kho qhov muag ntawm 45℃lub kaum sab xis mus rau qhov chaw kho qhov muag. Excitation light yog reflected rau hauv lub hom phiaj lens thiab tsom mus rau tus qauv. Cov fluorescence tsim los ntawm cov qauv, nrog rau lub teeb pom kev zoo tshwm sim los ntawm qhov chaw ntawm lub hom phiaj lens thiab npog, ib txhij nkag mus rau lub hom phiaj lens thiab rov qab mus rau lub dichroic beamsplitter, uas cais lub teeb excitation ntawm fluorescence. Qhov seem excitation teeb yog ces absorbed los ntawm lub barrier lim.
Los ntawm swapping lim blocks nrog sib txawv ua ke ntawm excitation lim, dichroic beamsplitter thiab barrier lim, qhov yuav tsum tau rau txawv fluorescent tshuaj tiv thaiv cov khoom yuav ua tau raws li. Qhov zoo ntawm hom fluorescence microscope no suav nrog kev teeb pom kev zoo hauv cheeb tsam, cov duab ntse, thiab ua kom cov fluorescence siv ntau dua.
