Dab tsi yog Cov Txheej Txheem Tshwj Xeeb ntawm Fluorescent Microscope?
Cov xim lim thaiv yog ib feem tseem ceeb ntawm lub tshuab hluav taws xob fluorescence, thiab nws cov khoom tseem ceeb muaj xws li thawj qhov thaiv qhov thaiv rau lub teeb pom kev zoo, lub lim dej thib ob rau lub teeb pom kev zoo, thiab lub teeb pom kev sib cais. Cov xim lim qauv thiab cov npe ntawm ntau lub tuam txhab feem ntau tsis sib haum.
1. Excitation light filter thiab emission light filter: Raws li cov yam ntxwv ntawm lub teeb ci thiab cov xim fluorescent, cov nram qab no peb hom kev sib txuam feem ntau yog xaiv los muab lub teeb pom kev zoo nyob rau hauv ib qho kev sib tw wavelength thiab tso cai rau cov fluorescence zoo siab los ntawm cov qauv kom dhau mus thiab ncav cuag lub qhov muag rau cov duab.
UV excitation: Lub teeb pom kev zoo tuaj yeem tso cai rau UV teeb hla thiab thaiv lub teeb pom kev siab tshaj 400nm los ntawm kev hla. Qhov sib thooj emission lub teeb lim tso cai rau lub teeb xiav hla dhau, thiab lub teeb nyob rau hauv qhov chaw pom tshwm xiav, xws li thaum siv rau DAPI staining.
Blue light excitation: Lub teeb pom kev zoo tuaj yeem tso cai rau lub teeb xiav hla thiab thaiv lub teeb los ntawm lwm cov wavelengths. Qhov sib thooj emission lub teeb lim tso cai rau lub teeb ntsuab hla, xws li GFP staining markers.
Green light excitation: Lub excitation light filter tso cai rau lub teeb ntsuab hla thiab thaiv lub teeb los ntawm lwm cov wavelengths. Lub teeb pom kev sib raug zoo feem ntau tso cai rau lub teeb liab hla, xws li Rhodamine staining.
2. Semi pob tshab thiab semi reflective xim lim: Nws txoj haujlwm yog txhawm rau thaiv lub teeb pom kev zoo kom dhau los thiab cuam tshuam nws; Thiab emit lub teeb nyob rau hauv qhov sib thooj wavelength range. Nws cov qauv sib raug rau lub teeb lim excitation thiab lub emission lub teeb lim.
(2) Lub hom phiaj lens thiab eyepiece
Ntau lub hom phiaj lo ntsiab muag tuaj yeem siv tau, tab sis nws yog qhov zoo tshaj plaws los xaiv cov lo ntsiab muag nrog cov nplai ntxiv thiab chromatic aberration txo, vim tias lawv tus kheej fluorescence tsawg heev thiab lawv lub teeb kis tau zoo (nruab nrab wavelength) yog haum rau fluorescence. Vim lub fact tias cov fluorescence brightness ntawm ib tug duab nyob rau hauv lub microscope teb yog ncaj qha proportional rau lub square ntawm lub aperture piv ntawm lub hom phiaj lens thiab inversely proportional rau nws magnification, thiaj li yuav txhim kho lub brightness ntawm fluorescence dluab, lub hom phiaj lens nrog ib tug loj aperture ratio yuav tsum tau siv. Tshwj xeeb tshaj yog rau cov qauv uas tsis txaus fluorescence, lub hom phiaj lens nrog lub siab aperture ratio thiab siab transmittance yuav tsum tau siv, nrog rau ib tug eyepiece nrog qhov tsawg tshaj plaws magnification tau.
(3) Lwm yam khoom siv kho qhov muag
Cov txheej txheej ntawm daim iav feem ntau yog coated nrog txhuas vim tias txhuas absorbs ultraviolet tsawg dua thiab pom lub teeb nyob rau hauv lub xiav xiav cheeb tsam, reflecting tshaj 90% (thaum nyiaj muaj ib tug reflectivity ntawm tsuas yog 70%). Feem ntau, cov iav tiaj tus yog siv. Lub tsom iav tsom iav, tshwj xeeb tsim thiab tsim rau fluorescence microscopes, yog ua los ntawm quartz iav los yog lwm yam iav uas xa cov teeb ci ultraviolet. Lub teeb pom kev poob qis, ntxiv rau qhov muaj lub luag haujlwm ntawm lub teeb pom kev zoo, yog qhov tsim nyog rau kev soj ntsuam ncaj qha ntawm opaque thiab semi pob tshab qauv, xws li cov phaj tuab, lim membranes, kab mob kab mob, kab lis kev cai, thiab lwm yam qauv. Nyob rau hauv xyoo tas los no, ntau yam tshiab ntawm fluorescence microscopes tau tsim los siv lub teeb poob, hu ua lub teeb fluorescence microscope.
(4) Lub teeb ci
Niaj hnub no, siab - siab mercury teeb ntawm 50 lossis 100W feem ntau yog siv los ua lub teeb ci. Thaum lub sijhawm ua haujlwm, kev tso tawm tshwm sim ntawm ob lub electrodes, ua rau cov mercury evaporate thiab lub siab nyob rau hauv lub sphere kom sai (cov txheej txheem no feem ntau yuav siv li 5-15 feeb). Thaum lub sij hawm tus txheej txheem no, lub teeb quanta raug tso tawm, thiab lub wavelength ntawm lub teeb tso tawm yog txaus los excite ntau yam fluorescent. Yog li ntawd, nws yog dav siv nyob rau hauv fluorescence microscopes.
Lub neej kev pab cuam ntawm mercury teeb yog luv luv, feem ntau 200 teev. Nyob rau hauv teb rau qhov kev txwv no hauv kev pab cuam lub neej, nyob rau hauv xyoo tas los no, ib hom tshiab ntawm fluorescent teeb qhov chaw X-Cite tau dav siv nrog lub teeb lub neej ntev ntev ntawm 2000 teev thiab siv tau yooj yim - tsis tas yuav tsum tau preheating, npaj siv.
