Qhov txawv ntawm Fluorescence Microscopes thiab Ordinary Light Microscopes
Fluorescence microscopes txawv ntawm cov pa microscopes. Hloov chaw ntawm kev soj ntsuam cov qauv nrog lub teeb los ntawm lub teeb pom kev zoo tib yam, lawv siv lub teeb ntawm ib lub wavelength tshwj xeeb (feem ntau yog ultraviolet lossis xiav-violet lub teeb) los ua kom cov khoom siv fluorescent nyob rau hauv cov qauv ntawm lub tshuab tsom iav, ua rau cov tshuaj no tawm fluorescence. Yog li ntawd, lub teeb qhov chaw ntawm lub fluorescence microscope tsis ncaj qha illuminate tus qauv; Nws ua raws li lub zog los ua kom muaj zog cov tshuaj fluorescent endogenous hauv cov qauv. Peb tuaj yeem pom cov qauv tsis yog vim lub teeb pom kev los ntawm lub teeb pom kev ntawm nws tus kheej, tab sis vim tias cov tshuaj fluorescent hauv cov qauv nqus lub zog excitation lub zog thiab tom qab ntawd tawm fluorescence.
Nws ua raws li qhov tseem ceeb ntawm lub fluorescence microscope yog tias nws lub teeb ci tuaj yeem muab lub teeb pom kev zoo ntawm ib lub wavelength band, ua rau cov tshuaj fluorescent hauv cov tshuaj ntsuam xyuas kom tawm fluorescence ntawm txaus siv. Lub caij no, lub tshuab hluav taws xob fluorescence yuav tsum tau nruab nrog lub tshuab lim sib raug. Nws yog ib qho tseem ceeb ntsuas rau fluorescence histochemistry. Lub tshuab ntsuas fluorescence feem ntau yog tsim los ntawm lub teeb pom kev siab siab, lub tshuab lim dej (xws li cov lim dej lim dej thiab cov lim dej thaiv), lub tshuab kho qhov muag thiab cov duab thaij duab. Nws ua haujlwm los ntawm kev siv lub teeb ntawm ib lub wavelength tshwj xeeb los ua kom muaj cov tshuaj fluorescent.
① Cov txheej txheem ntawm Fluorescence Excitation Raws li lub wavelength ntau yam ntawm lub teeb excitation, muaj ob hom: UV excitation method (ultraviolet illumination method) thiab BV excitation method (blue-violet illumination method).The UV excitation method siv ultraviolet light shorter for 400 nm. Nws tsim cov fluorescence tsis pom, yog li cov fluorescence pom los ntawm cov qauv nws tus kheej. Txoj kev no yooj yim distinguishes tshwj xeeb fluorescence los ntawm autofluorescence ntawm keeb kwm yav dhau.
② BV Excitation Method The BV excitation method siv lub teeb excitation xws li violet mus rau xiav nyob nruab nrab ntawm 404 nm thiab 434 nm. Txij li thaum lub teeb xiav yog siv los irradiate cov qauv, lub excitation lim system rau txoj kev no yuav tsum tau thaiv lub teeb xiav thiab tsuas yog tso cai rau lub teeb ntsuab thiab daj kom dhau mus.Rau fluorescent antibody staining, qhov siab tshaj plaws nqus wavelength ntawm lub teeb excitation yog ze rau qhov siab tshaj plaws emission wavelength ntawm fluorescent zas xim. Vim li no, lub lim teeb rau BV excitation yuav tsum siv lub lim txiav tawm. Txoj kev no siv lub teeb xiav rau excitation, yog li cov fluorescence sau efficiency yog qhov siab thiab cov duab ci tuaj yeem tau txais. Nws qhov tsis zoo yog tias fluorescence siab tshaj 500 nm tsis tuaj yeem kuaj pom, thiab tsuas yog cov duab daj-ntsuab tuaj yeem pom saum 500 nm. Hauv fluorescent antibody staining, kev txiav txim siab ntawm qhov tshwj xeeb tso siab rau cov ntaub ntawv xim, yog li qhov tsis zoo ntawm BV excitation txoj kev tuaj yeem yog ib qho kev txwv loj thaum tham txog kev hloov maj mam.
