Qhov sib txawv thiab cov yam ntxwv ntawm fluorescence microscope thiab zoo tib yam optical microscope
Fluorescence microscope yog txawv los ntawm cov khoom siv kho qhov muag zoo tib yam. Nws tsis soj ntsuam cov qauv los ntawm qhov illumination ntawm lub teeb pom kev zoo tib yam. Hloov chaw, nws siv lub teeb ntawm qee qhov wavelength (feem ntau yog lub teeb ultraviolet, xiav-violet lub teeb) los ua kom cov khoom siv fluorescent hauv cov qauv hauv qab lub tshuab tsom, ua rau nws tawm fluorescence. Yog li, Lub teeb ci ntawm lub tshuab hluav taws xob fluorescence tsis ua haujlwm ncaj qha, tab sis raws li lub zog hluav taws xob uas ua rau cov khoom siv fluorescent sab hauv ntawm cov qauv. Qhov laj thawj yog vim li cas peb tuaj yeem soj ntsuam cov qauv tsis yog vim lub teeb pom kev ntawm lub teeb, tab sis qhov tshwm sim fluorescence tshwm sim los ntawm cov khoom siv fluorescent hauv cov qauv nqus lub zog lub zog. Nws tuaj yeem pom tias tus yam ntxwv tseem ceeb ntawm lub tshuab hluav taws xob fluorescence yog tias nws lub teeb pom kev tuaj yeem muab ntau lub teeb pom kev zoo hauv qhov tshwj xeeb wavelength, kom cov tshuaj fluorescent hauv cov qauv hauv kev kuaj xyuas tuaj yeem tau txais qhov tsim nyog siv lub teeb pom kev zoo. Nyob rau tib lub sijhawm, fluorescence microscopes yuav tsum muaj cov tshuab lim zoo sib xws. Fluorescence microscopy yog ib qho cuab yeej tseem ceeb rau fluorescence histochemistry. Nws yog tsim los ntawm ultra-high-pressure teeb qhov chaw, lim system (xws li excitation thiab suppression lim daim hlau), optical system thiab photography system. Nws siv lub teeb ntawm ib lub wavelength los ua kom cov qauv kom tawm fluorescence.
1. Txoj hauv kev ua kom muaj zog fluorescence: Raws li lub wavelength ntau ntawm lub teeb, nws muab faib ua ob hom: UV excitation method (siv ultraviolet illumination method) thiab BV excitation method (siv lub teeb xiav-violet). Txoj kev UV excitation siv ze-ultraviolet lub teeb luv dua 400nm rau excitation. Tsis muaj lub teeb pom kev zoo nyob rau hauv txoj kev no, yog li qhov pom fluorescence qhia tau hais tias muaj cov fluorescence ntawm cov dye, ua kom yooj yim rau kev paub qhov txawv ntawm cov fluorescence ntawm cov qauv ntawm autofluorescence ntawm cov ntaub so ntswg tom qab.
2. BV excitation method: excitation los ntawm ultraviolet mus rau xiav lub teeb nyob rau hauv 404nm thiab 434nm. Txoj kev no siv lub teeb xiav los ua kom pom cov qauv, yog li qhov kev txiav tawm lim ntawm fluorescence kev soj ntsuam yuav tsum siv lub lim uas tuaj yeem thaiv lub teeb xiav thiab tag nrho dhau qhov xav tau ntsuab thiab daj fluorescence. Fluorescent dyes siv nyob rau hauv txoj kev fluorescent antibody. Qhov siab tshaj plaws nqus wavelength ntawm excitation lub teeb thiab lub siab tshaj plaws emission wavelength ntawm fluorescence yog nyob ze, yog li cov lim siv nyob rau hauv BV excitation txoj kev yuav tsum tau siv ib tug ntse cutoff lim. Txoj kev no tuaj yeem siv lub teeb xiav ua lub teeb pom kev zoo, yog li qhov kev nqus ntawm cov xim fluorescent yog siab dua thiab cov duab ci tuaj yeem tau txais. Qhov tsis zoo yog tias fluorescence qis dua 500nm tsis tuaj yeem pom, thaum fluorescence saum 500nm ua rau tag nrho cov duab pom daj. Hauv txoj kev fluorescent antibody, qhov tshwj xeeb ntawm cov xim fluorescent feem ntau yog txiav txim los ntawm nws cov xim tshwj xeeb. Yog li ntawd, thaum sib tham txog qhov hloov maj mam, qhov tsis txaus ntawm cov lus hais saum toj no ntawm BV excitation txoj kev feem ntau muaj kev cuam tshuam zoo.
