Koj Paub Ntau npaum li cas txog Fluorescence Microscopy
Fluorescence microscopes feem ntau siv cov roj teeb uas muaj mercury uas siv los ua kom lub teeb pom kev zoo. Cov ntxaij lim dej yog siv los lim tawm lub teeb tsis xav tau, tawm hauv tsuas yog lub teeb siab, lub teeb ntshiab uas ua rau cov fluorophore. Tom qab lub teeb monochromatic irradiates tus qauv los ntawm lub hom phiaj lens, tus qauv yuav zoo siab rau emit lub teeb (fluorescence), thiab ob qho tib si fluorescence thiab lub teeb excitation yuav rov qab los ntawm txoj kev kho qhov muag ntawm lub hom phiaj lens. Hauv qhov no, daim iav dichroic yuav tsum tau lim lub teeb pom kev zoo. , cia tsuas yog cov fluorescence peb yuav tsum pom los ntawm.
Qhov fluorescence no mus txog qhov muag ntawm lub qhov muag ntawm txoj kev teeb ntawm lub tshuab tsom, thiab tom qab ntawd nkag mus rau peb lub qhov muag, qhov chaw uas peb tuaj yeem pom cov fluorescence tawm los ntawm fluorophore.
Kev kuaj xyuas ua ntej thiab hloov kho ntawm fluorescence microscope:
(1) Ua ntej txhua qhov kev soj ntsuam fluorescence, nws yog ib qho tsim nyog yuav tsum tau ua tib zoo saib xyuas cov filament alignment, optical path focus, aperture diaphragm, thiab field diaphragm settings of the fluorescence device.
(2) Txawm hais tias qhov xav tau fluorescence excitation / emission lim los ua ke tau raug ntsia rau hauv lub converter, seb lub fluorescence microscope lub hom phiaj lens puas raug teeb tsa, thiab tshem tawm cov roj stains thiab plua plav ntawm lub lens pem hauv ntej ntawm lub hom phiaj lens.
(3) Yog hais tias theem sib piv kev soj ntsuam ntawm lub teeb kis tau zoo nyob rau tib lub sijhawm, nws yuav tsum tau kuaj xyuas qhov sib txuas ntawm qhov nruab nrab ntawm lub condenser thiab theem sib piv ntawm lub nplhaib rov qab rau lub hom phiaj lens.
(4) Txheeb xyuas seb tus qauv cab kuj (slide iav, npog iav thiab lwm yam khoom siv) puas yog cov kua lossis hmoov av, thiab seb lub thickness puas nyob rau hauv qhov kev ntsuas ua haujlwm nyob deb ntawm lub hom phiaj lens. Cov qauv txiav yuav tsum tsis txhob tuab heev, nyiam dua tsawg dua lossis sib npaug li 10 μm.
(5) Vim hais tias lub teeb pom kev zoo muaj cov kab hluav taws xob ultraviolet, lub teeb xim av-shielding phaj yog muab tso rau saum lub hauv ntej ntawm theem kom tiv thaiv ultraviolet rays los ntawm kev puas tsuaj rau lub retina.
(6) Lub zog tsis ruaj khov yuav txo qhov kev pab cuam lub neej ntawm lub teeb hluav taws xob siab mercury, thiab lub teeb ci hluav taws xob tau nruab nrog lub zog ruaj khov.
(7) Txhawm rau ua kom lub neej ntev ntawm lub teeb mercury, nws tuaj yeem muab tua 15 feeb tom qab nws qhib; Thaum lub hwj chim fluorescent ntawm lub teeb mercury raug kaw, nws yuav tsum tau tos tsawg kawg 10 feeb kom rov pib lub mercury vapor kom txias thiab rov qab mus rau qhov qub, txwv tsis pub lub neej ntawm lub teeb yuav raug cuam tshuam.
Kev soj ntsuam duab los ntawm fluorescence microscope:
(1) Txog 5-10 feeb tom qab tig lub teeb fluorescent, qhov kev siv ntawm lub teeb excitation zoo li ruaj khov, thiab cov qauv raug thauj khoom rau kev soj ntsuam; Txhawm rau tiv thaiv cov fluorescence quenching ntawm cov qauv tshwm sim los ntawm lub teeb excitation ntau dhau thaum lub sij hawm ntawm kev tsom thiab nrhiav cov khoom, ua ntej zoom tawm lub fluorescence microscope Kho lub teeb excitation mus rau nruab nrab siv nrog lub aperture diaphragm los yog ntxiv ND lim, thiab txav tus qauv theem tsis tu ncua. Tom qab paub meej tias daim iav duab, kho rau lub xeev fluorescent rau kev tua thiab kaw.
(2) Kev hloov kho rau cov duab tsis zoo. Ntxiv nrog rau cov qauv kev npaj, cov kev hloov kho tsim nyog uas tuaj yeem ua tau yog:
① Tsis suav nrog cov khoom siv tiv thaiv lub teeb los yog lub teeb-txhim kho cov khoom siv hauv cov duab kho qhov muag, xws li DIC accessories, ND lim, thiab lwm yam.
②Readjust the receiver focus and aperture diaphragm size of the fluorescence microscope.
③ Ua tib zoo kho qhov sib txawv kho lub nplhaib ntawm fluorescence microscope lub hom phiaj lens.
Daim ntawv thov cov ntsiab lus ntawm Fluorescence Microscopy
Fluorescence microscopy siv "actinic fluorescence" duab. Yog hais tias qhov kev xaiv excitation wavelength yog nyob ze-ultraviolet cheeb tsam (320-400nm), uas yog pom tsis tau ntawm lub qhov muag liab qab, lub emission spectrum ntawm fluorescence kuj luv dua li qhov nruab nrab wavelength ntawm lub teeb pom kev zoo ib yam li daim iav qhov chaw. txhim kho. High-zog photons sib tsoo nrog electrons, ua rau cov electrons hloov ntawm hauv av mus rau lub xeev zoo siab. Cov hluav taws xob hauv lub xeev zoo siab heev tsis ruaj khov thiab yuav poob rov qab rau hauv av. Hauv cov txheej txheem no, ib feem ntawm lub zog thermal yuav raug siv thiab cov photons tshiab yuav raug tso tawm. Lub photon tshiab muaj lub zog qis dua qhov qub photon thiab yog li ntawd muaj lub wavelength ntev dua. Txij li cov photon wavelength tshiab txawv ntawm photon wavelength ntawm qhov teeb meem teeb pom kev zoo, ob lub teeb ntawm lub teeb nrog cov wavelengths sib txawv tau sib cais los ntawm qee qhov kev ua haujlwm kho qhov muag, yog li ntawd peb tsuas pom cov emission tshiab photons (fluorescence teeb liab), uas yog, fluorescence microscope pom cov duab fluorescence.
