Daim ntawv thov ntawm Fluorescence Total Internal Reflection Fluorescence Microscopy
TIRFM (Tag nrho Internal Reflection Fluorescence Microscope), tag nrho cov sab hauv reflection fluorescence microscope, thaum lub teeb nkag mus rau hauv nruab nrab nrog ib tug qis refractive index los ntawm ib tug nruab nrab nrog ib tug siab refractive index, yog hais tias qhov teeb meem lub kaum sab xis loj txaus, tag nrho cov teeb yog reflected yam tsis muaj refraction, Tab sis nyob rau hauv lub interface ntawm ob lub xov xwm tsim evanescent tsis muaj zog uas yuav excite lub fluorescence nyob rau hauv 100nm nyob ze ntawm lub interface kom paub qhov kev soj ntsuam ntawm cov khoom nto. Lub teeb pom kev zoo tuaj yeem raug xa mus los ntawm lub teeb pom kev ntawm lub teeb pom kev zoo fluorescence microscope lossis lub teeb pom kev tshwj xeeb, thiab qhov xwm txheej ntawm lub laser tuaj yeem tswj tau. Txoj kev ua kom nrawm nrawm nrawm yog siv los tiv thaiv lub teeb pom kev zoo los ntawm kev nkag mus rau lub ntes. Lub teeb excitation ntawm qhov sib txuas ntawm iav thiab dej ua rau tag nrho sab hauv siv los ntawm kev xav. Vim lub exponential attenuation ntawm lub excitation lub teeb, tsuas yog cov qauv cheeb tsam ze rau tag nrho cov reflection nto yuav tsim fluorescence reflection, uas zoo heev txo qhov cuam tshuam ntawm lub teeb tom qab lub suab nrov rau lub hom phiaj soj ntsuam, yog li no technology yog dav siv nyob rau hauv lub dynamic soj ntsuam. ntawm cov khoom ntawm tes.
Schematic daim duab ntawm tag nrho cov internal reflection fluorescence microscopy (TIRFM)
①Sample ②Evanescent yoj ntau ③Cov iav ④Oil immersion ⑤Target ⑥Emission beam (cim) ⑦ Excitation beam
Yuav kom ua tiav tag nrho cov kev xav sab hauv, yuav tsum muaj qhov xwm txheej loj, piv txwv li, lub kaum sab xis ntawm lub iav-dej interface yog ntau dua 61 degrees. Qhov no tuaj yeem ua tiav los ntawm prism, hu ua prism-based TIRFM, los yog los ntawm lub hom phiaj lens nrog lub siab tus lej aperture, uas yog hu ua hom phiaj-hom TIRFM. Tam sim no kev lag luam tag nrho sab hauv kev xav txog fluorescence microscopes yog feem ntau ntawm lub hom phiaj lens hom, nrog kev kub ceev thiab siab precision.
Tag nrho cov kev xav sab hauv fluorescence microscopy yog siv dav hauv qee qhov chaw lom neeg vim tias nws tuaj yeem paub txog kev soj ntsuam fluorescence nyob rau hauv qhov sib txawv heev (tsawg dua 100nm) ntawm qhov chaw ntawm cov khoom. Xws li cov kev siv hauv qab no:
Kev soj ntsuam ntawm cov duab ntawm tes: cell membrane nto qauv, cell nto hu, membrane nto dynamics/protein localization.
Kev soj ntsuam ib leeg-molecule thiab manipulation: myosin, actin, thiab Cy3-labeled ATP.
Cell membrane nto txav: xws li vesicle engulfment, vesicle exhalation, thiab vesicle exocrine. tus
Kev soj ntsuam ntawm calcium sparking phenomenon nyob rau hauv cell membrane, ion channel saib xyuas.
Kev tshawb fawb molecular motors: rotational motors, cytoskeletal proteins, polymers, G proteins, ring proteins, nucleotide motors.
Ntxiv rau thaj tsam ntawm biology, nws kuj muaj cov ntawv thov zoo hauv kev ua haujlwm ntawm chemistry rau kev soj ntsuam cov qauv tshuaj molecular.
