photobleachingle fluorescence nashtam

koshangalkkum stharangalkkumullile thanmaathrakalude chalanam parisodhikkaan upayogikkunna oru fluorescence microscopy saankethikathayaanu photobleachingile fluorescence nashtam (flip). oru kosha stharathe saadhaaranayaayi nireekshanatthinu anuvadikkunnathinu fluorescent dye upayogichu label cheiyunnu. eee label cheytha bhaagatthinte oru prathyeka pradesham oru konfokkal lazer scanning microscopinte beam upayogichu niravadhi thavana bleach cheiyunnu. oro imaging scan cheythathinusheshavum bleaching veendum sambhavikkunnu. bleach cheyyaatha fluoroforukal bleach cheytha fluoroforukalaayi kaimaattam cheyyappedunnathinaal axus cheyyaavunna ellah fluoroforukalum bleach cheyyappedunnuvennu urappakkaan ithu niravadhi thavana sambhavikkunnu, ithu kosha stharathiloode chalanathinu kaaranamaakunnu. koshatthil mothatthil photobleachinginte phalangal nirnnayikkunnathin aaa pradeshathu ninnulla fluoresensinte alavu oru nischitha kaalayalavil alakkunnu.
pareekshanathmaka sajjeekaranam.
photobleaching sambhavikkunnathin munbu, koshangalkk oru fluorescent protein kuthivaykkanam, palappozhum oru green fluorescent protein (jii. eph. pii), ithu targettucheytha proteinukale fluorese cheyyaan anuvadikkukayum athinaal prakriyayiludaneelam pinthudarukayum cheyyum. appol, thaalpparyamulla oru mekhala nirvachikkanam. thaalpparyamulla eee prarambha mekhalayil saadhaaranayaayi muzhuvan koshamo niravadhi koshangalo adangiyirikkunnu. maruvashathu, photobleaching thaalpparyamulla pradeshathinu purathaanu sambhavikkunnath; athinaal oru photobleaching mekhalayum nirvachikkendathundu. moonnaamathe mekhala, alakkal nadakkunna pradeshavum nirnnayikkendathundu. photobleachinginu munbu fluorescence nirnnayikkaan niravadhi prarambha scanukal nadathendathundu. eee scanukal niyanthrana skaanukalaayi varthikkum, athumaayi photobleach cheytha scanukal pinneedu thaarathamyam cheyyum. appol photobleaching sambhavikkam. oro bleach pulsinum idayil, fluorescent metteeriyal veendedukkaan samayam anuvadikkendathu aavasyamaanu. kooduthal padanathinaayi oro bleach pulsinum sesham udan thanne thaalpparyamulla pradeshatthinte niravadhi scanukal edukkunnathum pradhaanamaan. thaalpparyamulla mekhalayile fluorescencile maattam moonnu vazhikalil onnil alakkaan kazhiyum. imej settukalude drishya parisodhanayude adisthaanathil thaalpparyamulla pradeshangalude sthaanam, valuppam, ennam enniva thiranjedukkuka ennathaanu ettavum saadhaaranamaayath. vyathyastha probe mobilittiyude mekhalakal vyakthigatha imej adisthaanathil kandethunnathiloodeyo chalikkunna vasthukkalil ninnulla fluorescence nashtatthinte physical modalingiloodeyo aanu mattu rund, puthiyathum ennaal kooduthal vishwasaneeyavumaaya sameepanagal.
fluorescence nashtappedunnathu nirvachikkunnath fluorescent label cheytha proteininte chalanathmaka bhinnasankhyayo allengil photobleached eriyayilekku veendedukkaan kazhivulla fluoroforukalude bhinnasankhyayo aanu. fluorescencinte apoornnamaaya nashtam soochippikkunnathu bleach cheytha pradeshathekku neengukayo sanjarikkukayo cheyyaatha fluoroforukal undennaanu. ithu fluorescent-label cheytha proteinukalude nischalamaaya bhinnasamkhyaye allengil photobleached eriyayilekku veendedukkaan kazhiyaatha fluoroforukalude bhinnasamkhyaye nirvachikkaan anuvadikkunnu. koshatthinte backie bhaagangalil ninnu adachitta compartmentukalil proteinukal undennu chalanaseshi soochippikkunnu, ithu aavarthichulla photobleaching avare baadikkunnathil ninnu thadayunnu.
aaplikkeshanukal.
membranus oorganellukalude thudarcha parishodhikkunnu.
membranus oorgaanellukalude thudarcha nirnnayikkuka ennathaanu flippinte praadhamika upayogam. thaalpparyamulla mekhalayile fluoresensinte alavu nireekshichaanu eee thudarchayo abhaavamo nirnnayikkunnath. fluorescence poornnamaayum nashtappettaal, ithu oorganellukal thudarchayaayirikkunnave soochippikkunnu. ennirunnaalum, fluorescence poornnamaayi nashtappettaal, oorganellukalkkidayil thudarcha undaakilla. pakaram, eee oorganellukal compartmentalise cheythathinaal eathenkilum photobleached fluoroforukalude kaimaattatthinaayi adaykkunnu. golgi upakaranam, endoplasmic rettikulam, nucleus ennivayude thudarcha flip upayogichu parishodhichittundu.
nucleus cytoplasavum thammilulla vinimaya nirakk.
cytoplaasathil ninnu nucleusilekku proteinukal engane shuttle cheyyappedunnuvennu nirnnayikkukayum thudarnnu eee shuttle sambhavikkunna nirakk nirnnayikkukayum cheyyuka ennathaanu flippinte mattu rund, pathivayi upayogikkappedaatha upayogangal. eathu bhagangal ulppettirikkunnuvennu avaa eppol shuttle prakriyayil ulppedunnuvennum nirnnayikkaan, thudarchayaaya scanukal nireekshikkunnu. shuttle prakriyayil cytoplaasathinte oru bhagam ethrayum vegam upayogikkunnu, athra vegathil athu fluorescence poornnamaayum nashtappedutthunnu. eee prakriyayude thathfalamaayundaakunna chithram poornnamaayum photoblicade cytoplasm aayirikkanam. nucleusil ninnu cytoplaasathilekkal nuclear kayattumathiyilum koshavum pankedukkukayaanenkil, nucleusil photobleaching sambhavikkum.
nucleus cytoplasavum thammilulla vinimaya nirakkum itharathilulla daattayil ninnu nirnnayikkaan kazhiyum. eee sandarbhangalil, photobleachinginte pradesham nucleusinullilaanu. shuttle athivegathil sambhavikkukayaanenkil, edutha frameukaliloode nuclear compartmentinulli fluorescence level athivegam kurayum. ennirunnaalum, shuttle pathukke sambhavikkukayaanenkil, fluorescence level baadhikkappedaathe thudarum allengil cheruthaayi kurayum.
flip vezhsas. pongacham.
flip palappozhum upayogikkunnu, ithu photobleachinginu (frap) sesham fluorescence veendedukkalumaayi adutha bandhamullathaan. eee rund microscopy saankethikavidyakalum thammilulla pradhana vyathyasam, orotta photobleaching sambhavathin sesham veendedukkaanulla oru koshatthinte kazhivinekkurichulla patanam frappil ulppedunnu ennathaanu, athesamayam onniladhikam photobleaching sambhavangalkk sesham fluorescence nashtam koshatthiludaneelam engane vyaapikkunnu ennathinekkurichulla patanam flippil ulppedunnu. uddeshyathile eee vyathyasam koshatthinte eathu bhagangal nireekshikkunnu ennathilum vyathyaasathilekku nayikkunnu. fraapil, yadhaarthathil photobleach cheytha mekhalayaanu thaalpparyamulla mekhala. neremarichu, photoblich cheiyunna pradeshathinu purathaanu thaalpparyamulla pradesham. mattoru pradhana vyathyasam, fraapil, oru photobleaching ivantum fluoroforukal ethra nannayi bleach cheytha sthalathekku madangunnu ennu nireekshikkaan oru veendedukkal kaalayalavum undu ennathaanu. ennirunnaalum, maruvashathu, bleaching pradeshathekku bleach cheyyaatha fluoroforukal madangunnathu thadayaan onniladhikam photobleaching sambhavangal sambhavikkunnu. flip pole, membranus oorgaanellukalude thudarchayekkurichulla padanatthil frap upayogikkunnu. jii. eph. pii. tagg cheytha proteinukalude chalanathmakatha nirnnayikkaan flip frap palappozhum orumichu upayogikkunnu. chalana nirakk pariganikkathe oru koshatthinte pradeshangal thammilulla tanmatra kaimaattam alakkaanum flip upayogikkaam. oru koshatthinullile protein kadathinekkurichu kooduthal samagramaya vishakalanam nadatthaan ithu anuvadikkunnu. photobleaching mathramulla pradeshangalile proteinukalude chalanathmakatha nirnnayikkaan praadhamikamaayi upayogapradamaaya frappil ninnu ithu vyathyasthamaan.
saadhyathayulla sankeernathakal.
flippilum fraapilum ulppedunna niravadhi sankeernathakalundu. rund tharatthilulla microscopykalum jeevanulla koshangale parishodhikkunnathinaal, koshangal neengaanulla saadhyatha ellaayppozhum undu, ithu thettaaya falangalkku kaaranamaakunnu. ithu ozhivakkaanulla ettavum nalla maargam oru alainment algoritam upayogikkuka ennathaanu, ithu eathu chalanathinum nashtapariharam nalkukayum chalanam moolamulla pishakinte valiyoru bhagam illaathaakkukayum cheyyum. eee pareekshanangalil, phalangal krameekarikkunnathinum fluorescencile mothatthilulla nashtatthinu veendedukkal vakram sariyaakkunnathinum oru niyanthrana group undaayirikkunnathum pradhaanamaan. pishak kuraykkunnathinulla mattoru maarggam photobleaching orotta pradeshatho pradeshatho nilanirthuka ennathaanu. photobleaching moolamulla fluorescence nashtavumaayi thaarathamyappeduthumbol foto-kedupaadukal moolamundaakunna fluorescence nashtam niyanthrikkunnathinum parimithappeduthunnathinum eee parimiti sahayikkum.

