Cov qauv tshwj xeeb ntawm fluorescence microscope suav nrog:
(I) Xim lim system
Xim lim yog ib qho tseem ceeb ntawm lub fluorescence microscope, nws cov khoom tseem ceeb los ntawm lub teeb pom kev zoo lim (thawj lub teeb ci), emission lub teeb lim (thib ob barrier lim) thiab ib nrab-pob tshab thiab ib nrab-tiv thaiv lim (beam-splitting mirror) muaj pes tsawg leeg . Ntau cov tuam ntxhab ntawm cov xim lim qauv, lub npe feem ntau tsis sib xws.
1. Excitation lub teeb lim thiab emission lub teeb lim: raws li lub teeb ci thiab cov yam ntxwv ntawm fluorochrome, feem ntau xaiv peb hom kev sib txuam hauv qab no, muab qee qhov wavelength ntau ntawm lub teeb excitation, thiab ua rau cov qauv excitation ntawm fluorescence los ntawm eyepiece rau daim duab.
UV excitation: Lub excitation lub teeb lim tso cai rau kev sib kis ntawm UV teeb thiab thaiv qhov kev pom ntawm lub teeb pom kev siab tshaj 400nm. Cov lim dej sib thooj tso cai rau kev hla ntawm lub teeb xiav, uas tshwm xiav hauv qhov pom, xws li DAPI staining.
Blue light excitation: Lub excitation lub teeb lim tso cai rau lub teeb xiav hla dhau, thaiv lwm cov wavelengths ntawm lub teeb. Cov lim dej sib thooj tso cai rau kev hla lub teeb ntsuab, piv txwv li rau GFP staining markers.
Green excitation: Lub lim dej tso cai rau lub teeb ntsuab hla dhau, thaiv lwm cov wavelengths. Cov lim dej sib thooj feem ntau tso cai rau lub teeb liab hla dhau, xws li Rhodamine staining.
2. Ib nrab-pob tshab thiab ib nrab-reflective lim: nws lub luag hauj lwm yog kom tag nrho cov excitation ntawm lub teeb los ntawm lub reflection; thiab los ntawm qhov sib thooj wavelength ntau ntawm lub teeb emitted. Nws cov qauv thiab cov excitation ntawm lub teeb lim thiab emission ntawm lub teeb lim sib raug rau.
(B) Lub hom phiaj lens thiab eyepiece
Muaj ntau lub hom phiaj lo ntsiab muag tuaj yeem siv tau, tab sis * kev xaiv zoo ntawm symmetrical, achromatic lub hom phiaj lo ntsiab muag, vim tias nws cov fluorescence yog me me thiab lub teeb kis tau tus mob (nruab nrab wavelength) haum rau fluorescence. Raws li cov fluorescence brightness ntawm daim duab nyob rau hauv lub microscope ntawm saib yog proportional rau lub square ntawm lub hom phiaj lub muzzle tus nqi thiab inversely proportional rau nws magnification, thiaj li yuav txhim kho lub brightness ntawm cov duab fluorescence, lub hom phiaj nrog ib tug loj muzzle npaum li cas yuav tsum tau. siv. Tshwj xeeb tshaj yog rau cov fluorescence tsis muaj zog txaus cov qauv, yuav tsum tau siv lub qhov ncauj loj npaum li cas, siab lub teeb kis tau tus mob lub hom phiaj, nrog rau tsawg li tsawg tau nrog eyepiece.
(C) lwm yam khoom siv kho qhov muag
Reflector, nws cov txheej txheej feem ntau yog txhuas-plated, vim tias txhuas ntawm ultraviolet thiab pom lub teeb pom kev ntawm xiav-violet cheeb tsam tsawg dua, xav txog li 90% lossis ntau dua (thaum lub zog ntawm cov nyiaj tsuas yog 70%). Flat reflectors feem ntau yog siv. tsom teeb tsom iav, tsim thiab tsim tshwj xeeb rau fluorescence microscopy tsom teeb tsom iav ua los ntawm quartz iav los yog lwm yam UV-pob tshab iav. Poob lub teeb khoom, uas ntxiv rau kev ua haujlwm ntawm lub teeb kis tau tus mob, tsim nyog rau cov opaque thiab translucent specimens, xws li tuab zaj duab xis, lim membrane, colony, cov ntaub so ntswg kab lis kev cai thiab lwm yam qauv rau kev soj ntsuam ncaj qha. Nyob rau hauv xyoo tas los no, txoj kev loj hlob ntawm ib hom tshiab ntawm fluorescence microscope siv lub teeb poob, hu ua ntog fluorescence microscope.
(D) lub teeb ci
Niaj hnub no, 50 lossis 100W high-pressure mercury teeb yog siv los ua lub teeb ci. Ua hauj lwm los ntawm ob lub electrodes tawm, tshwm sim los ntawm mercury evaporation, lub pob ntawm huab cua siab nce ceev (cov txheej txheem no feem ntau yuav siv sij hawm li 5 ~ 15min), thiab nyob rau hauv tus txheej txheem ntawm emission ntawm lub teeb quanta, lub wavelength ntawm lub tso tawm lub teeb yog txaus los excite. Txhua hom tshuaj fluorescent, yog li ntawd, feem ntau siv rau fluorescence microscopy.
Mercury teeb muaj lub neej ua haujlwm luv luv, feem ntau yog 200hrs, thiab teb rau qhov kev txwv lub neej no, nyob rau xyoo tas los no, lub teeb ci fluorescent tshiab, X-Cite, tau siv dav rau nws lub neej ntev ntawm 2000hrs thiab nws siv tau yooj yim. - tsis tas yuav tsum tau preheating, thiab nws yog npaj los siv txoj cai tawm ntawm lub thawv.
