Imakroskopu ye-fluorescence luhlobo lwemakroskopu yamehlo esebenzisa umthombo wokukhanya ophezulu ukukhanyisa isampuli kunye nokuvuselela ii-fluorochromes kwisampulu. Ukukhanya komzekelo ngokuqhelekileyo kwenziwa ngomthombo wokukhanya okhupha ukukhanya kwe-ultraviolet. Zisetyenziswa ngokubanzi kwiinkalo zebhayoloji, ezonyango kunye nezoshishino.
Isebenza njani imakroskopu ye-fluorescence?
I-microscope ye-fluorescence isebenzisa i-mercury okanye isibane se-xenon ukuvelisa ukukhanya kwe-ultraviolet. Ukukhanya kungena kwi-microscope kwaye kubethe isibuko se-dichroic - isibuko esibonisa uluhlu olulodwa lwamaza obude kwaye sivumela olunye uluhlu ukuba ludlule. Isibuko se-dichroic sibonisa ukukhanya kwe-ultraviolet ukuya kwi-specimen. Eminye imizekelo ye-fluoresce ngokwemvelo phantsi kokukhanya kwe-ultraviolet kuba iqulethe izinto ezikhanyayo ezifana ne-chlorophyll. Ukuba umfuziselo oza kujongwa awukho ngokwemvelo, unokungcoliswa ngedayi ekhanyayo ebizwa ngokuba yi-fluorochromes.
Iquka ntoni imakroskopu ye-fluorescence?
1. Umthombo wokukhanya
Eyona mithombo yokukhanya ixhaphakileyo yimercury, xenon, kunye neeLED. IMercury ibonelela ngowona mgangatho wokukhanya ubalaseleyo kwimakroskopu ye-fluorescence. Ii-LED zithandwa kakhulu ngenxa yokuba zibiza kakhulu kuneminye imithombo kwaye zisebenzisa amandla amancinci.
| Isibane seMercury | Isibane seXenon | I-LED | |
| Ubude beWaveleng bokuKhanya okuThunyiweyo | 350-370 nm | 400nm ~ 450nm | 400-700 nm |
| Ubomi bonke(iyure) | 200-300 | 400-600 | 10000 |
| Inzuzo | 1. Ukukhanya okuPhezulu okuPhakamileyo, eyona mithombo yokukhanya iqhelekileyo. 2. Ikhupha i-ultraviolet enamandla kunye ne-blue-violet ukuze ivuse zonke iintlobo ze-fluorophores. 3. Imifanekiso eqaqambileyo nenemibala. | 1. Ubunzulu be-spectral buzinzile. 2. Ukuqina okubonakalayo okunamandla kwi-infrared kunye ne-mid-infrared. | 1. Kulula ukutshintsha. 2. Akukho mfuneko yokufudumala. 3. Ingavulwa kwaye icinywe ngoko nangoko. 4. Lawula amandla omthombo wokukhanya. |
| Ukusilela | 1. Ubomi obufutshane. 2. Ixesha elide lokufudumala. | 1. Ibhokisi yamandla ombane aphantsi okhethekileyo yeDC iyafuneka ukwenzela ubushushu obugqithisileyo. 2. Ixabiso eliphezulu kunesibane seMercury | 1. Isibane se-UV sibuthathaka kunesibane semercury. 2.Ubude bokukhanya ngabunye bufuna i-LED eyahlukileyo ngenxa yobubanzi obumxinwa ababukhuphayo. |
I-Mercury Fluorescent Attachment
I-LED Fluorescent Attachment
2. Isihluzo sokuKhuthaza
Isihluzo sovuselelo sibalulekile ekusebenzeni kwe-fluorescence microscope. Igqithisa ukukhanya kobude obufutshane obunokufunxwa yidayi ye-fluorescent. Kwakhona, ithintela eminye imithombo yokukhanya okunomdla.
Isihluzi Sokuhluza(B,G,U,V)
3. Isibuko seDichroic
Isibuko se-dichroic luhlobo lwesihluzo se-optical esibonisa ukukhanya kumaza athile ngelixa sihambisa ezinye. Isetyenziswa kwimakroskopu ye-fluorescence ukwahlula imincili kunye nokuphuma kwamaza amaza.
4. Isihluzo sokukhupha
Isihluzi sokukhutshwa sidlula kuphela ubude bamaza akhutshwa yi-fluorophore kwaye sivalela konke ukukhanya okungafunekiyo ngaphandle kweli bhanti - ngakumbi imincili yamaza.
5. Idayi zeFluorescent
Idayi eziFluorescent ziikhompawundi zezinto eziphilayo ezinepropathi ye-fluorescence, ezinokuthi ngayo zenze umfanekiso we-fluorescent ngokukhupha ukukhanya okuluhlaza okuhluke kakhulu okubonakalayo emva kokuvuyiswa kukukhanya okukhulu kwe-ultraviolet. Iidayi zefluorescent eziqhelekileyo ezisetyenziswayo zezi; I-DAPI (49,6-diamidino-2-phenylindole), i-acridine orange, i-auramine-rhodamine, i-Alexa Fluors, okanye i-DyLight 488.
Zingaphi iintlobo zemakroskopu ekhanyayo?
1. Imakroskopu ye-Epifluorescence ethe nkqo:
Lolona hlobo luxhaphakileyo lwe-fluorescence microscope. Ukuvuswa kwe-fluorophore kunye nokufunyanwa kwe-fluorescence kwenziwa ngendlela yokukhanya efanayo (okt ngenjongo). Uninzi lweemakroskopu ze-fluorescence, ngakumbi ezo zisetyenziswa kwisayensi yobomi, zezoyilo lwe-epifluorescence.
I-BS-2081F yoPhando ngeMikroskopu yebhayoloji eFluorescent
2. I-Confocal Fluorescence Microscope:
Imakroskopu yeConfocal Fluorescence: Olu hlobo lwemakroskopu ye-fluorescence ludibanisa ukuskena kwelaser kunye nokukhanya kwefluorescent ukuvelisa umfanekiso. Ingasetyenziswa kuluhlu olubanzi lwezicelo, ezinjengokufunda iiseli kunye nezicubu, ukufumanisa iiproteni kunye nezinye izinto ngaphakathi kweeseli, kunye nokulinganisa ubukhulu bezinto.
I-BS-2081F yoPhando ngeMikroskopu yebhayoloji eFluorescent
3. Imakroskopu yeFluorescence eNgenelweyo
Olu hlobo lomthombo wokukhanya we-microscope kunye ne-condenser zibekwe phezulu, zijonge ezantsi. I-angle yokukhanyisa kufuneka ibe kwi-90 degrees ngokubhekiselele kumphezulu we-specimen ehlolwayo.
I-BS-2095F iMikroskopu yeFluorescent eguqulweyo
Ziziphi iingenelo kunye nokungalunganga kweemicroscopes ze-fluorescence?
Izinto eziluncedo:
- Yenza ukuqwalaselwa kweeseli kwizinto eziphilayo ngaphandle kokuzonakalisa
-Ibonelela ngemifanekiso ephezulu enemibala echanekileyo
- Ivumela uphononongo lweenkqubo eziphilayo kwiiseli
- Isetyenziselwa ukuchonga iintlobo ezahlukeneyo zeemolekyuli ngaphakathi kweeseli, ezinjengeeprotheyini okanye ii-nucleic acid.
Izinto ezingeloncedo:
-Ivumela kuphela ukuqwalaselwa kwezakhiwo ezithile ngaphakathi kwiseli ephawulwe ngedayi ye-fluorescent.
-I-photobleaching ngenxa ye-electron excitation ngexesha lenkqubo ye-fluorescence inokuchaphazela iimolekyuli ezisebenzayo zedayi ze-fluorescent. Ngenxa yoko, iidayi ezisebenzayo zinokuphulukana nepropathi yazo yeekhemikhali zokuphuma kwe-fluorescence.
-Iiseli zichaphazeleka kwisiphumo se-phototoxic emva kokungcoliswa kwedayi ye-fluorescent, njengoko iimolekyuli ze-fluorophore zithatha iifotoni zamandla aphezulu ukusuka kukukhanya okufutshane kwamaza.
Yintoni usetyenziso lwemakroskopu ye-fluorescence?
Iimicroscopes zeFluorescence zisetyenziswa ngokubanzi kwiinkalo ezahlukeneyo zophando kunye nokusetyenziswa okubandakanya i-biochemistry, i-cell biology, i-microbiology, i-immunology, kunye neyeza.
1. Kwintsimi yebhayoloji, i-microscope ye-fluorescence yenza ukuba kuchongwe ngokuchanekileyo kunye neenkcukacha zamacandelo eselula kunye ne-submicroscopic kunye nemisebenzi ngoncedo lwe-labeling yedayi ye-fluorescent.
2. Kwicandelo lezonyango, i-microscope ye-fluorescence inokusebenzisa ii-reagents ze-fluorescent ukubona ubukho kunye nokusabalalisa ibhaktheriya kunye neentsholongwane, okanye ukuncedisa ekubhaleni iithagethi zotyando ukulungiselela utyando.
3. Kwintsimi ye-mineralogy, i-microscope ye-fluorescence isoloko isetyenziselwa ukufunda izinto ezineempawu ze-fluorescence ezizenzekelayo, ezifana ne-asphalt, i-petroleum, amalahle, i-graphene oxide kunye nezinye iiminerali.
4. Kwinzululwazi yezixhobo, imakroskopu ye-fluorescence ingasetyenziswa kushishino lwelaphu okanye kushishino lwephepha ukuhlalutya imathiriyeli esekwe kwifiber.
Ixesha lokuposa: Mar-14-2023