I-microscopy ye-Fluorescence iguqule amandla ethu okuba nomfanekiso ngqondweni kunye nokufunda iisampulu zebhayoloji, isivumela ukuba siphengulule kwihlabathi elintsonkothileyo leeseli kunye neemolekyuli. Icandelo eliphambili le-fluorescence microscopy ngumthombo wokukhanya osetyenziselwa ukuvuyisa iimolekyuli ze-fluorescent ngaphakathi kwesampulu. Ukutyhubela iminyaka, kuye kwasetyenziswa imithombo yokukhanya eyahlukahlukeneyo, nganye ineempawu zayo neengenelo zayo.
1. Isibane seMercury
Isibane semercury soxinzelelo oluphezulu, ukusukela kwi50 ukuya kuma-200 watts, sakhiwe kusetyenziswa iglasi yequartz kwaye singqukuva ngobume. Iqulethe umlinganiselo othile we-mercury ngaphakathi. Xa isebenza, ukukhutshwa kwenzeka phakathi kwee-electrode ezimbini, kubangela ukuba i-mercury iphume, kwaye uxinzelelo lwangaphakathi kwi-sphere lukhula ngokukhawuleza. Le nkqubo ithatha malunga nemizuzu emi-5 ukuya kweli-15.
Ukukhutshwa kwesibane esiphezulu se-mercury kubangela ukuchithwa kunye nokunciphisa iamolekyu ze-mercury ngexesha lokukhutshwa kwe-electrode, okukhokelela ekuphumeni kweefotoni ezikhanyayo.
Ikhupha i-ultraviolet eyomeleleyo kunye nokukhanya okuluhlaza okwesibhakabhaka, iyenza ilungele izinto ezahlukeneyo ezinomdla ze-fluorescent, yiyo loo nto isetyenziswa kakhulu kwi-fluorescence microscopy.
2. Izibane zeXenon
Omnye umthombo wokukhanya omhlophe osetyenziswa ngokuqhelekileyo kwi-fluorescence microscopy sisibane se-xenon. Izibane zeXenon, njengezibane zemercury, zibonelela ngobubanzi obubanzi bamaza amaza ukusuka kwi-ultraviolet ukuya kwi-infrared ekufutshane. Nangona kunjalo, ziyahluka kwi-spectra yazo yokuvuselela.
Izibane zeMercury zigxininisa ukukhutshwa kwazo kwindawo ekufutshane ne-ultraviolet, eluhlaza okwesibhakabhaka, kunye nohlaza, okuqinisekisa ukuveliswa kweempawu ezikhanyayo ze-fluorescent kodwa ziza ne-phototoxicity eyomeleleyo. Ngenxa yoko, izibane ze-HBO zihlala zigcinelwe iisampulu ezisisigxina okanye umfanekiso obuthathaka we-fluorescence. Ngokwahlukileyo, imithombo yesibane ye-xenon ineprofayili yokuvuselela egudileyo, evumela ukuthelekiswa kokuqina kumaza ahlukeneyo. Olu phawu luluncedo kwizicelo ezifana nemilinganiselo yoxinaniso lwe-calcium ion. Izibane zeXenon zikwabonisa imincili eyomeleleyo kuluhlu olusondeleyo lwe-infrared, ngakumbi malunga ne-800-1000 nm.
Izibane ze-XBO zineenzuzo zilandelayo ngaphezu kwezibane ze-HBO:
① Ubukhulu obufanayo bokubukeka
② Ukuqina okubonakalayo okunamandla kwimimandla ye-infrared kunye ne-mid-infrared
③ Imveliso enkulu yamandla, iyenza kube lula ukufikelela kwindawo yokuvula injongo.
3. Ii-LEDs
Kwiminyaka yakutshanje, umbangi omtsha uye wavela kwindawo yokukhanya kwe-fluorescence microscopy: ii-LED. Ii-LEDs zibonelela ngenzuzo yokutshintsha okukhawulezileyo kokucinywa kwii-milliseconds, ukunciphisa amaxesha okuvezwa kwesampulu kunye nokwandisa ubomi beesampulu ezibuthathaka. Ngaphaya koko, ukukhanya kwe-LED kubonakalisa ukubola okukhawulezayo kunye nokuchanekileyo, kunciphisa kakhulu i-phototoxicity ngexesha lokuvavanywa kwexesha elide leseli.
Xa kuthelekiswa nemithombo yokukhanya emhlophe, ii-LED zikholisa ukukhupha ngaphakathi kwembonakalo emxinwa yovuselelo. Nangona kunjalo, iibhendi ezininzi ze-LED ziyafumaneka, zivumela usetyenziso oluguquguqukayo lwemibala emininzi ye-fluorescence, zenza ii-LED zibe lukhetho oluya luthandwa kakhulu kwiiseti zanamhlanje ze-fluorescence microscopy.
4. Umthombo wokukhanya weLaser
Imithombo yokukhanya ye-Laser i-monochromatic kakhulu kwaye iyalathiswa, iyenza ibe yinto efanelekileyo kwi-microscopy enesisombululo esiphezulu, kubandakanywa neendlela zokulungiswa okuphezulu ezifana ne-STED (i-Stimulated Emission Depletion) kunye ne-PALM (i-Photoactivated Localization Microscopy). Ukukhanya kweLaser ngokuqhelekileyo kukhethwe ukufanisa ubude obuthile bokuvuselela obufunekayo kwithagethi ye-fluorophore, ebonelela ngokukhethwa okuphezulu kunye nokuchaneka kwi-excitation ye-fluorescence.
Ukukhethwa komthombo wokukhanya we-fluorescence microscope kuxhomekeke kwiimfuno ezithile zovavanyo kunye neempawu zesampulu. Nceda uzive ukhululekile ukuqhagamshelana nathi ukuba ufuna naluphi na uncedo
Ixesha lokuposa: Sep-13-2023