
fluorescence-jeevitakaala imaging microscopy

fluorescence-jeevitakaala imaging microscopy ledhaa flim anedi oka namuunaa nundi fluorophore yokka photan udgaaram yokka ghaataanka kshayam retulo vyatyaasaala aadhaaramgaa oka imaging saanketikata. dheenini confocal microscopy, too-photan excitation microscopy mariyu multifotan tomography imaging technicga upayoginchavacchu.
fluorophore yokka fluorescence jeevitakaalam (flt), daani teevrata kante, chitraanni anchulo srushtinchadaaniki upayoginchabadutundi. fluorescence jeevitakaalam fluorophore yokka sthaanika suukshma-vaataavaranampai aadharapadi untundi, tadwara kaanti muulam yokka prakaasamloo maarpu, nepathya kaanti teevrata ledhaa parimita photo-bleaching kaaranamgaa fluorescence teevratalo edainaa tappudu kolatalu nivaarinchabadataayi. eee saanketikata namuunaa yokka mandapati poralalo photan chedaragotte prabhavanni tagginche prayojanaanni kooda kaligi undhi. suukshma paryaavaranampai aadharapadi unnanduna, jeevitakaala kolatalu Ph, snigdhata mariyu rasayana jaatula yekagrataku suuchikagaa upayoginchabaddaayi.
fluorescence jeevitakaalam.
photan dwara uttejitamayye fluorophore aneka vibhina (radiative mariyu/ledhaa naan-radiative) kshaya maargaala dwara kshayam raetla aadhaaramgaa oka nirdishta sambhaavyatatoo naela sthithiki padipotundi. pratideeptini gamaninchadaaniki, eee margalalo okati photan yokka aakasmika udgaaram dwara undaali. samishti vivaranalo, vidudalayye fluorescence kalakramena ksheenistundi
ekkada
paina perkonna vaatilo, suutram _ 3 anedi samayam, suutram _ 4 anedi pratideepaka jeevitakaalam, suutram _ 5 anedi suutram _ 6 vadda prarambha pratideepakatvam, mariyu suutram _ 7 prati kshayam maargaaniki raetlu, veetilo kaneesam okati pratideepaka kshayam raetu suutram _ 8 ayi undaali. murry mukhyamgaa, jeevitakaalam, suutram _ 4 anedi prarambha teevrata mariyu vidudalayye kaanti nundi swatantramgaa untundi. rasayana samvedanalo teevrata laeni aadhaarita kolatalu cheyadaaniki dheenini upayoginchavacchu.
kolata.
pratideepti-jeevitakaala imaging suutram _ 4 dwara nirnayinchabadina prati pixel teevratatoo chitraalanu istundi, idhi ververu pratideepti kshayam raetlu (aaa padaarthaalu sarigga adhe tarangadairghyamto fluoreses ayinappatikee) unna padaarthaala madhya vyatyaasaanni chuudataaniki anumatistundi, mariyu itara kshayam margalalo maarpulanu choopinche chitraalanu kooda utpatti chestundi, fret imaging vantidi.
pulsed prakaasam.
pulsed sorsnu upayoginchadam dwara samaya domainlo fluorescence jeevitakaalaanni nirnayinchavacchu. 
fluoroforla janabha ultrashort ledhaa delta pulls dwara uttejitamainappudu, paina vivarinchina vidhamgaa samayam-parishkarinchabadina fluorescence vipareetamgaa ksheenistundi. ayithe, excitation pulls ledhaa detection response vistrutamgaa unte, kolichina fluorescence, d (t), puurtigaa ghatankamga undadu. vaayidya pratispandana function, irf (tii) anedi dickay function, eff (tii) thoo kalisipotundi ledhaa militam cheyabadutundi.
suutram _ 11
muulam, detector mariyu electronics yokka vaayidya pratispandananu saadhaaranamgaa chellacheduruga unna excitation light nundi kolavavacchu. denaminator sunnaki daggaragaa unnappudu frequency domainloni vibhajana adhika sabdaanni utpatti chestundi kabatti kshayam functionnu (mariyu sambandhita jeevitakaalaalu) punaruddharinchadam adanapu savaallanu kaligistundi.
tii. sii. ess. pii. sii.
samayam-paraspara sambandham gala yeka-photan lekkimpu (tii. sii. ess. pii. sii) saadhaaranamgaa upayoginchabadutundi endukante idhi muulam teevrata mariyu yeka photan pulls vyaapti loe vaividhyaalanu bharti chestundi. 
vaanijyaparamaina tii. sii. ess. pii. sii parikaraalanu upayoginchi pratideepti kshayam vakrarekhanu 405 eff. ess. varaku samaya spashtatato namodu cheyavachu. 
namodaina fluorescence kshayam histogram poison ganaamkaalaku kattubadi untundi, idhi amarchinapudu fitt yokka manchitanaanni nirnayinchadamlo pariganinchabadutundi. 
marinta pratyaekamgaa, excitation lazer pulls kuu sambandhinchi vegavantamaina single-photan detector (saadhaaranamgaa photo-multiplier tube (pmt) ledhaa single photan avalanch photo diode (spaad)) dwara vyaktigata photaanlanu gurtinche samayaalanu tii. sii. ess. pii. sii. namodu chestundi. 
recordinglu bahula lazer pulls kosam punaraavrutamavutaayi mariyu taginanta namodu cheyabadina sanghatanala tarvaata, eee namodu cheyabadina samaya binduvulannintilo sanghatanala sankhya yokka histograamnu nirminchagalugutaaru. 
eee histogram appudu aasakti yokka ghaataanka jeevitakaala kshayam functionnu kaligi unna ghaataanka functionku saripotundi, mariyu tadanugunamgaa jeevitakaala paraamitini velikiteeyavacchu. 
16 nundi 64 muulakaalatoe kuudina bahula-chaanal pmt vyavasthalu vaanijyaparamgaa andubaatulo unnaayi, ayithe iteevala pradarsinchabadina cmos single-photan avalanch diode (spaad)-tccpc flim vyavasthalu marinta ekkuva sankhyalo detection chaanellanu mariyu adanapu takkuva kharchutho kuudina empikalanu andinchagalavu.
gating paddhati.
pulls excitation ippatikee eee paddhatiloo upayoginchabadutundi. pulls namuunaanu cherukune mundu, kontha kaanti dicroic addam dwara pratibimbistumdi mariyu photodiode dwara gurtinchabadutundi, idhi sisidi detector mundu kuurchunna gated optical intensifier (goyi) nuu niyantrinche delay generatornu sakriyam chestundi. alasyam tarvaata terichinappudu kontha samayam matrame gurtinchadaaniki goyi anumatistundi. anduvalla, sardubaatu cheyagala aalasya generatortho, namuunaa yokka pratideepti kshayam yokka samaya paridhini kaligi unna aneka aalasya samayala tarvaata pratideepti udgaaraanni sekarinchagalugutaaru. iteevali samvatsaraalalo samagramaina teevramaina sisidi kemeraalu marketloki pravesinchaayi. eee cameralalo image intensifier, sisidi sensor mariyu integrated delay generator untaayi. 200 ps varaku athi takkuva gating samayalu mariyu 10 ps aalasya dasalu kaligina ii. sii. sii. dii kemeraalu sub-nanosecond resolution flimnu anumatistaayi. endoscopto kalipi eee saanketikata medadu kanitula intraoperative nirdhaaranaku upayoginchabadutundi.
dasha modulation.
pratideepti jeevitakaalaanni frequency domainlo faze-modulation paddhati dwara nirnayinchavacchu. eee paddhati led, diode lazer ledhaa electro-optic modulator ledhaa acousto-optic modulatortho kalipi nirantara taranga muulam vanti adhika pounahpunyamlo (500 emhechz varaku) pulls ledhaa modulate cheyabadina kaanti vanarunu upayogistundi. pratideepti anedi (a.) demodulated mariyu (bii.) dasha maarchabadindi; rendu parimaanaalu fluorophore yokka lakshana kshayam samayalaku sambandhinchinavi. alaage, excitation mariyu fluorescence saine taramgaalaku y-bhagalu modulate cheyabadataayi mariyu eee y-bhagala modulation nishpatti nundi jeevitakaalaanni nirnayinchavacchu. anduvalla, jeevitakaalaaniki 2 viluvalanu faze-modulation paddhati nundi nirnayinchavacchu. eee prayogaatmaka paaraamitula amarika vidhaanaala dwara jeevitakaalam nirnayinchabadutundi. pmt-aadhaarita ledhaa camera-aadhaarita frequency domine flim yokka prayojanam emitante, daani vegavantamaina jeevitakaala chitra samuparjana, idhi pratyaksha kana parisoedhana vanti anuvartanaalaku anukuulamgaa untundi.
vislaeshanha.
vislaeshanha algorithm yokka lakshyam kolichina kshayam nundi swachchamaina kshayam vakrarekhanu velikiteesi, jeevitakaalam (lanu) anchanaa veyadam. rendodi saadhaaranamgaa oke ledhaa bahula ghaataanka vidhulanu amarchadam dwara saadhinchabadutundi. eee samasyanu parishkarinchadaaniki vividha paddhatulu abhivruddhi cheyabaddaayi. atyanta vistrutamgaa upayoginche saanketikata athi takkuva chadarapu punaraavruta punah-parivartana, idhi avasheshaala yokka baruvutho kuudina mottaanni tagginchadam meeda aadharapadi untundi. eee saanketikatalo saiddhaantika ghaataanka kshayam vakratalu parikara pratispandana functiontho vankaragaa untaayi, idhi vidigaa kolustaaru, mariyu kaneesa kanugonabade varaku vividha inputla kosam avasheshaala punaraavruta ganana dwara uttama amarika kanugonabadutundi. time binn i loe fluorescence signal yokka pariseelanala suutram _ 12 kosam, jeevitakaala anchanaa kanishteekarinchadam dwara nirvahinchabadutundih
suutram _ 13
tarangadairghyam aadhaarita parikara pratispandana functiontho sahaa prayogaatmaka ibbandulatoe paatu, punaraavruta dii-convolution samasya yokka ganita chikitsa neerugaa munduku saagadu mariyu idhi nemmadigaa jarige prakriya, idhi flim prarambha rojullo pixel-bye-pixel vislaeshanhaku asadhyamga maarindi. 
naan fitting paddhatulu aakarshanheeyamgaa untaayi endukante avi jeevitakaala anchanaaku chaalaa vegavantamaina parishkaranni andistaayi. eee vargamlooni pradhana mariyu suutigaa unde paddhatullo okati vegavantamaina jeevitakaala nirdhaarana (rld) paddhati. rld kshayam vakrarekhanu samaana vedalpu suutram _ 14t yokka rendu bhagaluga vibhajinchadam dwara jeevitakaalaanni mariyu vaati vyaaptulanu neerugaa lekkistundi. kshayam vakrarekhanu samaana samaya vyavadhilo suutram _ 14t loe ekikrutam cheyadam dwara vislaeshanha nirvahinchabadutundih
suutram _ 16
i anedi i-th chaanello namodu cheyabadina signal mariyu k anedi chaanella sankhya. jeevitakaalaanni eee krindi vaatini upayoginchi anchanaa veyavachuhu
suutram _ 17
bahula ghaataanka kshayam kosam eee sameekaranam sagatu jeevitakaalaanni andistundi. dvi-ghaataanka kshayala vislaeshanhaku eee paddhatini vistarinchavacchu. eee paddhati yokka oka pradhana lopam emitante, idhi parikara pratispandana prabhavanni parigananaloki teesukodu mariyu eee kaaranamgaa kolichina kshayam vakrarekhala yokka prarambha bhaganni vislaeshanhalaloo vismarinchaali. dheeni ardham signal loe kontha bhaagam paaraveyabadindani mariyu takkuva jeevitakaalaanni anchanaa vaeyadaaniki khachitatvam taggutundani.
convolution siddhaantam yokka aasaktikaramaina lakshanaalalo okati, convolution yokka samagrata anedi samagratanu yerparuchukune kaarakaala yokka utpatti. kolichina vakrarekha nundi swachchamaina kshayam vakrarekhanu tirigi pondadaaniki eee lakshanaanni upayoginchukune konni paddhatulu roopaantaram chendina pradaesamloe panicheestaayi. laplace mariyu forier parivartana, laguerre gaas vistaranatoe patuga, roopaantaram chendina pradaesamloe jeevitakaalaanni anchanaa vaeyadaaniki upayoginchabaddaayi. eee vidhanalu deconvalution aadhaarita paddhatula kante vaegamgaa untaayi, conny avi kattirimpu mariyu namuunaa samasyalatho baadhapadataayi. antekakunda, lagur gaas vistarana vanti paddhatula anuvartanam ganitasaastraparamgaa sanklishtamgaa untundi. forier paddhatulalo oke ghaataanka kshayam vakrarekha yokka jeevitakaalam ilaa ivvabadutundihi
suutram _ 18
ekkadah
suutram _ 19
mariyu n anedi harmonic sankhya mariyu t anedi gurtinche mottam samaya paridhi.
applications.
kanaalu mariyu kanitulalo photosensitizerlanu gurtinchadaaniki alaage nishpattilo imaging kashtamgaa unna sandarbhaallo aandolana chendadaaniki oka paddhatigaa jeevasaastramlo flim pradhanamgaa upayoginchabadindi. 
eee saanketikata 1980la chivaralo mariyu 1990la praarambhamlo abhivruddhi cheyabadindi (gating paddhatih bugil mariyu itarulu. 1989. 1989 loe, dasha modulation lakoviz ett all. 1992,) 1990la chivaralo marinta vistrutamgaa vartimpajeyabadataaniki mundu. sell kalcharlo, idhi egf grahaka sanketam mariyu akrama ravaanaanu adhyayanam cheyadaaniki upayoginchabadindi. 
nuclear envalap vadda vibhina homopolymerlalo rendu rakaala nuclear intermediate filament proteins laminulu eah mariyu bii 1 yokka paraspara charyanu choopinchadaaniki time domine flim (tidiflim) kooda upayoginchabadindi, ivi adhika kramam nirmaanaalalo okadaanitoe okati marinta sankarshana chendutaayi. neuronlalo flim imaging mukhyamgaa upayogapadutundi, ikkada medadu kanajaalam dwara kaanti chedaragottadam anedi nishpattilo imaging kosam samasyaatmakamgaa untundi. neuronlalo, pulsed prakaasam upayoginchi flim imagingnu raas, kaamki, rock mariyu runn family proteinlanu adhyayanam cheyadaaniki upayoginchaaru. clinical multifotan tomography intradermal cancer kanaalatoe paatu pharmaceutical mariyu cosmetic sammelanaalanu gurtinchadaaniki flim upayoginchabadindi.
iteevala mokkala kanaalalo flavanolsnu gurtinchadaaniki kooda flim upayoginchabadindi.
autofluorescent koenjaimlu naad (pii) hetch mariyu fyaad.
ksheerada gvakriyalo maarpulaku gurtulugaa koenjaimla nundi auto-fluorescensenu gurtinchadaaniki multy-photan flim ekkuvagaa upayoginchabadutundi.
bhayapette imaging.
endukante fluorophore yokka fluorescence jeevitakaalam radiative (i.ii. fluorescence) mariyu naan-radiative (i.ii. challarchadam, aandolana) prakriyalu, daata anuvu nundi sweekarta anuvuku shakti badilee daata jeevitakaalaanni taggistundi. 
anduvalla, flim upayoginchi pora kolatalu fluorophore yokka sthithulu/parisaraala madhya vivaksha chuupadaaniki oka paddhatini andistundi. 
teevrata-aadhaarita fret kolatalaku viruddhamgaa, flim-aadhaarita fret kolatalu kooda fluoroforla gaadataku sunnitamgaa undavu mariyu tadwara namuunaa antataa ekagrata mariyu udgaara teevratalo vaividhyaala dwara pravesapettina kalakhandalanu filter cheyagalavu.