Kev siv Inverted Fluorescence Microscope kom raug
Inverted fluorescence microscope yog ib lub tshuab siv ntau yam siv hauv kev tshawb fawb txog kev tshawb fawb txog roj ntsha siab kawg. Nws cov qauv yog complex thiab kim. Yog li ntawd, txhawm rau kom cov tub ntxhais kawm tiav kev kho mob kom raug kev ua haujlwm thiab siv cov tshuab hluav taws xob rov qab fluorescence, cov kws qhia ntawv yuav tsum xub ua kom pom qhov ua haujlwm tiag tiag ntawm lub tshuab hluav taws xob inverted fluorescence. Ua ntej, ib chav kawm tau tsim los kawm cov kev paub yooj yim ntawm inverted fluorescence microscopes, qhia cov qauv, cov ntsiab lus, kev ua haujlwm thiab cov kev siv yooj yim ntawm inverted fluorescence microscopes, thiab tom qab ntawd hauv kev ua haujlwm tshwj xeeb, rau cov qauv sib txawv, ua qauv sib txawv, los ntawm qhov ntiav. kom sib sib zog nqus, maj mam ua qauv siv inverted fluorescence microscopes los ntawm cov tub ntxhais kawm kho mob.
1. Kev siv qhov pom kev kaj lug zoo. Ib qho ntawm kev ua haujlwm ntawm inverted fluorescence microscope yog kev soj ntsuam qhov kaj. Lub hauv paus ntsiab lus tseem ceeb yog tias tom qab cov qauv yog irradiated los ntawm lub teeb, lub teeb haum ntawm txhua feem yog sib txawv kom paub cov duab nyob rau hauv ib tug ci keeb kwm yav dhau. Nws yog tsuas yog haum rau stained los yog xim hlwb, Kev soj ntsuam ntawm cov qauv xws li cov ntaub so ntswg.
2. Kev siv kom raug ntawm kev soj ntsuam theem sib piv. Qhov thib ob muaj nuj nqi ntawm lub inverted fluorescence microscope yog theem sib piv soj ntsuam. Nws lub hauv paus ntsiab lus yog los hloov txoj kev kho qhov muag sib txawv ntawm qhov sib txawv ntawm cov qauv mus rau qhov sib txawv amplitude. Kev soj ntsuam yog pom tau los ntawm ib theem sib piv lub hom phiaj lens nrog lub phaj phaj thiab lub condenser lens nrog lub nplhaib diaphragm. Nws yog tsuas yog siv tau rau kev soj ntsuam ntawm cov cell nyob los yog cov ntaub so ntswg tsis huv, nyob rau hauv cov txheej txheem ntawm kev sim qhia, ntxiv rau kom raug coj mus kuaj cov tub ntxhais kawm tiav kev soj ntsuam cov txheej txheem kev khiav hauj lwm ntawm theem sib piv kev soj ntsuam, nws yog ib qho tseem ceeb ntxiv rau coj lawv mus rau. nkag siab thiab paub cov qauv siv rau kev soj ntsuam theem sib txawv. Rau lawv, kev soj ntsuam microscopic ntawm cov kab lis kev cai yog feem ntau siv hauv kev tshawb fawb yav tom ntej. Ua ntej tshaj plaws, rau kev xaiv cov khoom siv ntawm tes, feem ntau pom zoo kom siv iav thiab yas kab lis kev cai tais diav, kab lis kev cai daim phiaj thiab kab lis kev cai fwj. Lawv feem ntau yog xim tsis muaj xim thiab pob tshab, thiab tuaj yeem pom ncaj qha rau ntawm lub tshuab hluav taws xob fluorescent inverted yam tsis muaj dej ntws tawm ntawm tes thiab lwm yam kev kho mob. Qhov thib ob, thaum soj ntsuam microscopic, tsis txhob qhib lub hau ntawm cov cuab yeej ntawm lub siab, thiaj li tsis txhob kis kab mob. Thaum kawg, rau cov neeg ua haujlwm, lawv yuav tsum tau hnav cov hnab looj tes, qhov ncauj qhov ntswg thiab cov tsho dawb, uas tuaj yeem tsis tsuas yog tiv thaiv lawv tus kheej kev noj qab haus huv, tab sis kuj tiv thaiv cov kab mob ntawm tes.
3. Kev siv cov kev soj ntsuam fluorescence kom raug. Qhov thib peb ua haujlwm ntawm lub tshuab hluav taws xob fluorescence inverted yog kev soj ntsuam fluorescence. Lub hauv paus ntsiab lus tseem ceeb yog siv lub teeb ci kom zoo siab rau cov qauv los tsim cov fluorescence, kom paub qhov kev soj ntsuam ntawm cov duab thiab txoj hauj lwm ntawm tus qauv. Nws yog qhov tsim nyog rau kev soj ntsuam cov qauv fluorescently sau npe. Kev soj ntsuam fluorescence yog ib qho tseem ceeb ntawm lub tshuab hluav taws xob inverted fluorescence, thiab nws cov khoom siv yog kim thiab tshwj xeeb siv. Txhawm rau kom cov tub ntxhais kawm tiav kev kho mob kom raug kev ua haujlwm thiab siv kev soj ntsuam fluorescence, cov kws qhia ntawv yuav tsum tsis yog qhia lawv cov kev ua haujlwm yooj yim, tab sis tseem ceeb tshaj, qhia lawv kom paub nws qhov tshwj xeeb. Ua ntej tshaj plaws, hauv kev npaj cov qauv, cov iav swb thiab npog iav yuav tsum muaj lub ntsej muag du thiab cov tuab tsim nyog (lub iav swb yuav tsum nyob nruab nrab ntawm 0.8mm thiab 1.2mm, thiab cov iav npog yuav tsum yog li ntawm { {5}}.17mm); cov qauv yuav tsum muaj qhov tsim nyog tuab thiab zam cov hlwb Overlapping lossis npog los ntawm impurities; qhov nruab nrab nruab nrab yuav tsum siv qhov sib npaug ntawm qhov sib npaug ntawm cov glycerol thiab carbonate buffer (0.5mol/L pH9.0-9.5). Qhov thib ob, ua ntej kev soj ntsuam fluorescent, lub teeb hluav taws xob hauv chav tsev thiab lub teeb dawb ntawm lub tshuab hluav taws xob fluorescent yuav tsum raug muab tua kom ntseeg tau tias ib puag ncig hauv chav tsaus. Thaum kawg, nyob rau hauv kev soj ntsuam fluorescence, lub sij hawm soj ntsuam yuav tsum nruj me ntsis tswj, feem ntau 1-2 teev / sij hawm yog tsim nyog, txwv tsis pub cov fluorescence yuav attenuate thiab ploj; Thaum kho qhov teeb pom kev zoo, hnav cov tsom iav tiv thaiv kom tsis txhob muaj kev puas tsuaj ntawm ultraviolet rays rau cov tsom iav.
4. Kev siv cov duab thaij kom raug. Hauv kev tshawb fawb tshawb fawb, nws tsis yog tsuas yog xav tau tias lub tshuab hluav taws xob rov qab fluorescence tuaj yeem paub ntau yam kev soj ntsuam, tab sis kuj xav kom lub tshuab hluav taws xob inverted fluorescence tuaj yeem paub cov duab ntes. Lub inverted fluorescence microscope system ua ke nrog lub tshuab ntsuas qhov muag nrog lub khoos phis tawj los ntawm kev hloov pauv photoelectric, tsis yog rau kev soj ntsuam microscopic ntawm lub qhov muag, tab sis kuj rau lub sijhawm tiag tiag dynamic duab saib, kho thiab txuag hauv lub computer.
