Fluorescence microscopy yasintha luso lathu lotha kuona ndi kuphunzira zamoyo, zomwe zatilola kuti tifufuze za dziko lovuta kwambiri la maselo ndi mamolekyu. Chigawo chachikulu cha microscopy ya fluorescence ndi gwero la kuwala lomwe limagwiritsidwa ntchito kusangalatsa mamolekyu a fulorosenti mkati mwa zitsanzo. Kwa zaka zambiri, magetsi osiyanasiyana akhala akugwiritsidwa ntchito, chilichonse chili ndi makhalidwe ake komanso ubwino wake.
1. Nyali ya Mercury
Nyali ya mercury yothamanga kwambiri, yoyambira 50 mpaka 200 watts, imapangidwa pogwiritsa ntchito galasi la quartz ndipo imakhala yozungulira. Lili ndi kuchuluka kwa mercury mkati. Ikagwira ntchito, kukhetsa kumachitika pakati pa maelekitirodi awiri, zomwe zimapangitsa kuti mercury isungunuke, ndipo kupanikizika kwamkati m'gawo kumawonjezeka kwambiri. Izi zimatenga pafupifupi mphindi 5 mpaka 15.
Kutuluka kwa nyali ya mercury yothamanga kwambiri kumabwera chifukwa cha kupasuka ndi kuchepetsedwa kwa mamolekyu a mercury panthawi yotulutsa ma elekitirodi, zomwe zimapangitsa kuti ma photon aziwala.
Imatulutsa kuwala kwamphamvu kwa ultraviolet ndi blue-violet, ndikupangitsa kuti ikhale yoyenera pazinthu zosiyanasiyana zosangalatsa za fulorosenti, chifukwa chake imagwiritsidwa ntchito kwambiri mu microscopy ya fluorescence.
2. Xenon Nyali
Gwero lina lomwe limagwiritsidwa ntchito moyera mu microscopy ya fluorescence ndi nyali ya xenon. Nyali za Xenon, monga nyali za mercury, zimapereka mawonekedwe ochulukirapo a mafunde kuchokera ku ultraviolet kupita kufupi ndi infrared. Komabe, amasiyana mu mawonekedwe awo osangalatsa.
Nyali za Mercury zimayang'ana kwambiri kutulutsa kwawo kumadera omwe ali pafupi ndi ultraviolet, buluu, ndi zobiriwira, zomwe zimatsimikizira kubadwa kwa ma siginecha owala koma amabwera ndi phototoxicity yamphamvu. Chifukwa chake, nyali za HBO nthawi zambiri zimasungidwa zitsanzo zokhazikika kapena zojambula zofooka za fluorescence. Mosiyana ndi izi, magwero a nyali za xenon ali ndi mbiri yosangalatsa yosangalatsa, zomwe zimalola kufananitsa mwamphamvu pamafunde osiyanasiyana. Khalidweli ndi lothandiza pakugwiritsa ntchito ngati miyeso ya ndende ya calcium ion. Nyali za Xenon zimawonetsanso chisangalalo champhamvu pafupi ndi infrared range, makamaka mozungulira 800-1000 nm.
Nyali za XBO zili ndi zabwino izi kuposa nyali za HBO:
① Kuwoneka kofananako kowoneka bwino
② Kuwoneka kwamphamvu kwambiri m'magawo a infrared ndi ma infrared
③ Kutulutsa mphamvu zambiri, kupangitsa kuti zikhale zosavuta kufikira pomwe mukufuna.
3. Ma LED
M'zaka zaposachedwa, wopikisana naye watsopano watulukira mu malo a fluorescence microscopy magwero kuwala: LEDs. Ma LED amapereka mwayi wosinthira mwachangu ma milliseconds, kuchepetsa nthawi yowonekera komanso kukulitsa moyo wa zitsanzo zofewa. Kuphatikiza apo, kuwala kwa LED kumawonetsa kuvunda mwachangu komanso molondola, kumachepetsa kwambiri toxicity panthawi yoyeserera kwanthawi yayitali ya cell.
Poyerekeza ndi kuwala koyera, ma LED nthawi zambiri amatulutsa mkati mwa sipekitiramu yocheperako. Komabe, magulu angapo a LED alipo, omwe amalola kugwiritsa ntchito mitundu yosiyanasiyana ya fluorescence, kupanga ma LED kukhala chisankho chodziwika bwino pamakonzedwe amakono a fluorescence microscopy.
4. Gwero la kuwala kwa laser
Magwero a kuwala kwa laser ndi monochromatic kwambiri komanso amawongolera, kuwapangitsa kukhala abwino kwa maikolosikopu apamwamba kwambiri, kuphatikiza njira zowunikira kwambiri monga STED (Stimulated Emission Depletion) ndi PALM (Photoactivated Localization Microscopy). Kuwala kwa laser kumasankhidwa kuti kufanane ndi kutalika kwake komwe kumafunikira pa fulorophore, kumapereka kusankha kwakukulu komanso kulondola kwachisangalalo cha fluorescence.
Kusankhidwa kwa gwero la kuwala kwa microscope ya fluorescence kumadalira zofunikira zoyesera ndi mawonekedwe a zitsanzo. Chonde khalani omasuka kulumikizana nafe ngati mukufuna thandizo lililonse
Nthawi yotumiza: Sep-13-2023