I-Fluorescence microscopy iguqule ikhono lethu lokubona ngeso lengqondo nokufunda izibonelo zebhayoloji, okusivumela ukuthi singene sijule ezweni eliyinkimbinkimbi lamaseli nama-molecule. Ingxenye ebalulekile ye-fluorescence microscopy umthombo wokukhanya osetshenziselwa ukujabulisa ama-molecule e-fluorescent ngaphakathi kwesampula. Phakathi neminyaka edlule, kuye kwasetshenziswa imithombo yokukhanya ehlukahlukene, ngayinye inezici zayo ezihlukile kanye nezinzuzo zayo.
1. Isibani seMercury
Isibani semercury esinengcindezi ephezulu, esisukela kumawathi angama-50 kuye kwangama-200, sakhiwe kusetshenziswa ingilazi yequartz futhi siyindilinga. Iqukethe inani elithile le-mercury ngaphakathi. Uma isebenza, kuphuma uketshezi phakathi kwama-electrode amabili, okwenza i-mercury ihwamuke, futhi ukucindezela kwangaphakathi ku-sphere kuyanda ngokushesha. Le nqubo ngokuvamile ithatha cishe amaminithi angu-5 kuya kwangu-15.
Ukukhishwa kwesibani se-mercury esinengcindezi ephezulu kubangela ukuhlakazeka nokuncipha kwama-molecule e-mercury ngesikhathi sokukhishwa kwe-electrode, okuholela ekukhishweni kwama-photon alula.
Ikhipha ukukhanya okunamandla kwe-ultraviolet kanye ne-blue-violet, okuyenza ifanelekele izinto ezihlukahlukene ezithokozisayo ze-fluorescent, yingakho isetshenziswa kakhulu ku-microscopy ye-fluorescence.
2. Amalambu eXenon
Omunye umthombo wokukhanya omhlophe ovame ukusetshenziswa ku-fluorescence microscopy isibani se-xenon. Amalambu e-Xenon, njengamalambu e-mercury, ahlinzeka nge-spectrum ebanzi yamaza wamaza ukusuka ku-ultraviolet kuya ku-infrared eduze. Nokho, ziyehluka nge-spectra yazo evusa amadlingozi.
Amalambu e-Mercury agxilisa ukukhishwa kwawo ezindaweni eziseduze ne-ultraviolet, eluhlaza okwesibhakabhaka, neluhlaza, okuqinisekisa ukukhiqizwa kwezimpawu ezikhanyayo ze-fluorescent kodwa eza ne-phototoxicity eqinile. Ngenxa yalokho, amalambu e-HBO ngokuvamile agcinelwe amasampula angashintshi noma i-imaging ye-fluorescence ebuthakathaka. Ngokuphambene, imithombo yesibani ye-xenon inephrofayili ye-excitation ebushelelezi, evumela ukuqhathaniswa kokuqina kumaza ahlukene. Lesi sici sinenzuzo ezinhlelweni ezifana nezilinganiso zokuhlushwa kwe-calcium ion. Amalambu e-Xenon aphinde abonise ukujabula okunamandla ebangeni le-infrared eseduze, ikakhulukazi eduze kuka-800-1000 nm.
Amalambu e-XBO anezinzuzo ezilandelayo ngaphezu kwamalambu e-HBO:
① Ukuqina kwe-spectral okufana okwengeziwe
② Ukuqina kwe-spectral okunamandla ezindaweni ze-infrared nemaphakathi ne-infrared
③ Ukukhipha amandla amakhulu, okwenza kube lula ukufinyelela indawo yomgomo.
3. Ama-LED
Eminyakeni yamuva nje, kuye kwavela umbangi omusha endaweni yemithombo yokukhanya ye-fluorescence microscopy: ama-LED. Ama-LED anikezela ngenzuzo yokushintsha okusheshayo kokucisha kuma-millisecond, ehlisa izikhathi zokuchayeka isampula kanye nokwelula ubude bempilo yamasampuli athambile. Ngaphezu kwalokho, ukukhanya kwe-LED kukhombisa ukubola okusheshayo nokunembayo, kunciphisa kakhulu ubuthi be-phototoxic ngesikhathi sokuhlolwa kweseli ephilayo yesikhathi eside.
Uma kuqhathaniswa nemithombo yokukhanya emhlophe, ama-LED ngokuvamile aphuma ngaphakathi kwe-spectrum yenjabulo emincane. Kodwa-ke, amabhande e-LED amaningi ayatholakala, avumela izinhlelo zokusebenza ze-fluorescence ezinemibala eminingi, okwenza ama-LED abe yinketho ethandwa kakhulu ekusetheni kwe-microscopy ye-fluorescence yesimanje.
4. Lasers Light Source
Imithombo yokukhanya ye-laser i-monochromatic kakhulu futhi iyaqondisa, iyenza ilungele imakroskopu yokulungiswa okuphezulu, okuhlanganisa namasu okulungiswa okuphezulu okufana ne-STED (Stimulated Emission Depletion) kanye ne-PALM (Photoactivated Localization Microscopy). Ukukhanya kwe-laser kuvame ukukhethwa ukuze kufane nobude beza begagasi besasasa obuthile obudingekayo ku-fluorophore eqondiwe, okunikeza ukukhetha okuphezulu nokunemba kwesasasa le-fluorescence.
Ukukhethwa komthombo wokukhanya wesibonakhulu se-fluorescence kuncike ezidingweni ezithile zokuhlola nezici zesampula. Sicela ukhululeke ukuxhumana nathi uma udinga noma yiluphi usizo
Isikhathi sokuthumela: Sep-13-2023