Qhov zoo thiab qhov tsis zoo ntawm Multiphoton Laser Scanning Microscopy
Multiphoton laser scanning microscopy yog ib txoj hauv kev sim ua raws li laser scanning microscopy thev naus laus zis, uas muab cov peev txheej kho qhov muag pom tseeb dua hauv kev soj ntsuam peb sab. Multiphoton fluorescence excitation method siv lub teeb liab ntev ntev los yog ze-infrared lub teeb los sau cov duab fluorescence siab ntawm cov qauv, uas muaj kev puas tsuaj tsawg heev rau cov qauv nyob, yog li nws tsim nyog rau cov duab nyob hauv cell, tshwj xeeb tshaj yog rau cov ntaub so ntswg nyob tuab xws li lub hlwb hlais, embryos, kev tshawb fawb ntawm tag nrho cov kabmob lossis txawm tias tag nrho cov kabmob.
Qhov zoo yog raws li nram no:
1. Zoo siab los ntawm lub teeb liab los yog lub teeb infrared, lub teeb tawg yog me me (qhov tawg ntawm cov khoom me me yog inversely proportional rau plaub lub zog ntawm lub wavelength).
2. Tsis tas yuav tsum muaj cov pinholes, thiab cov photons tawg ntau ntawm cov duab tuaj yeem sau tau.
3. Lub pinhole tsis tuaj yeem paub qhov txawv ntawm cov photons tawg tawm ntawm qhov chaw tawm ntawm lub ntsej muag lossis qhov chaw tsom mus, thiab qhov sib piv ntawm cov teeb liab-rau-suab nrov ntawm ntau cov duab duab hauv cov txheej sib sib zog nqus yog qhov zoo.
4. Lub teeb ci ultraviolet lossis pom pom siv rau ib leeg-photon excitation yog yooj yim absorbed thiab attenuated los ntawm cov qauv ua ntej lub beam mus txog lub focal dav hlau, thiab nws tsis yooj yim rau excite sib sib zog nqus txheej.
5. Nyob rau hauv cov nqe lus ntawm kev tshuaj ntsuam xyuas lub tshuab kuaj kab mob, thawj qhov kev txiav txim siab tsis yog ua kom lub cev ua haujlwm ntawm lub cev nws tus kheej, thiab tswj cov kev ncig ntawm dej, ion concentration, oxygen thiab cov as-ham. Nyob rau hauv cov ntaub ntawv ntawm lub teeb pom kev, ob qho tib si tshav kub thiab photon zog yuav tsum nyob rau hauv lub irradiation npaum li cas thiab lub teeb lub zog uas yuav tsis ua puas hlwb.
6. Multiphoton microscopy kuj muaj ntau yam zoo. Xws li kev daws teeb meem peb-dimensional, kev nkag mus tob, kev ua haujlwm tawg, lub teeb tom qab, teeb liab-rau-suab nrov piv, tswj, thiab lwm yam, muaj cov yam ntxwv zoo tshaj plaws uas laser microscopes tsis muaj lossis tsis muaj qhov sib piv yav dhau los.
Multiphoton confocal laser scanning microscopy tau txuas ntxiv mus rau ntau yam kev tshawb fawb thiab daim ntawv thov teb. Nws tuaj yeem ua qhov kev soj ntsuam peb-dimensional tsis muaj kev puas tsuaj ntawm cov qauv hauv lub xeev ntuj, thiab tuaj yeem txhim kho qhov kev daws teeb meem thiab teeb liab-rau-nruab nrab ntawm qhov system. Siv cov ntaub ntawv multiphoton excitation Nws tseem tuaj yeem paub peb-seem qhov siab cov ntaub ntawv khaws cia thiab peb-dimensional arbitrary kev taw qhia microfabrication, uas muaj daim ntawv thov siab. Nws ntseeg tau tias nrog kev txhim kho ntxiv ntawm cov tshuab, cov khoom siv, thiab cov tshuab laser ntsig txog multiphoton confocal microscopy, yuav muaj ntau dua Photon confocal laser scanning microscope yuav tau txais kev txhim kho ntau dua thiab siv dav dua.
Qhov tsis zoo yog raws li nram no:
1. Tsuas yog rau cov duab fluorescence xwb.
2. Yog tias cov qauv muaj xws li chromophores uas tuaj yeem nqus tau lub teeb pom kev zoo, xws li cov xim pigments, cov qauv yuav raug thermally puas.
3. Qhov kev daws teeb meem yog txo me ntsis. Txawm hais tias nws tuaj yeem txhim kho los ntawm kev siv lub confocal aperture tib lub sijhawm, lub teeb liab yuav ploj.
4. Vim muaj kev txwv ntawm kim ultrafast lasers, tus nqi ntawm multiphoton scanning microscope yog tus nqi siab.
