Fluorescence yog lub cev muaj zog tshwm sim los ntawm emission ntawm lub teeb lub zog thaum ib yam khoom yog illuminated los ntawm lub teeb los yog lwm yam ntaub ntawv ntawm electromagnetic tawg. Fluorescence microscopy yog ib qho khoom siv kho qhov muag uas siv cov fluorescence thiab phosphorescence nyob rau hauv situ los yog ua ke nrog kev xav thiab nqus los kawm cov khoom kho qhov muag ntawm cov khoom organic thiab inorganic. Fluorescence microscopy pab kom pom cov subcellular thiab sab hauv ntawm cov khoom kom meej meej. Nws tau pom nyob rau xyoo pua 20th thaum Lub Yim Hli Köhler, Carl Reichert thiab Heinrich Lehmann.
Tsab xov xwm index (nias mus dhia)
Yuav ua li casfluorescence microscopyua haujlwm
Fluorescence microscopy ua haujlwm
Fluorescence microscope Cheebtsam
1. Fluorophore
2. Lub teeb qhov chaw
3. Excitation lim
4.Dichroic daim iav
5. Tshem tawm lim
Qhov zoo ntawm Fluorescence Microscopy
Disadvantages ntawm fluorescence microscopy
Kev siv cov fluorescence microscopy
Yuav ua li cas fluorescence microscopy ua haujlwm
Fluorescence microscopy feem ntau ua haujlwm los ntawm staining cov muaj pes tsawg leeg thiab cov qauv ntawm cov qauv nrog dyes. Qhov no yog vim hais tias feem ntau ntawm cov cellular Cheebtsam tsis muaj xim nyob rau hauv cov xwm. Yog li ntawd, lawv tsis tuaj yeem yooj yim sib txawv ntawm ib leeg. Fluorophores los yog fluorescent molecules illuminated nrog luv wavelength lub teeb los yog electromagnetic rays yuav emit lub teeb. Lub teeb emitted muaj lub wavelength ntev dua piv rau qhov teeb meem tshwm sim. Feem ntau, lub sijhawm ntawm irradiation thiab emission yog ntsuas hauv nanoseconds thiab tuaj yeem tsis quav ntsej yooj yim. Lub teeb emission yog cais los ntawm qhov teeb meem teeb meem, uas qhia qhov tseeb qhov chaw ntawm fluorophore. Cov duab ntawm cov qauv yog tsim los ntawm lub teeb theem nrab es tsis yog qhov teeb meem thawj zaug.
Fluorescence microscopy ua haujlwm
Fluorescence microscopy ua haujlwm zoo ib yam li lub teeb microscopy. Nws muaj lub teeb pom kev zoo, lub lim dej tshwj xeeb bandpass thiab daim ntawv fluorescent ntawm cov qauv. Cov qauv uas yuav tsum tau kawm yog muab tso rau hauv lub tshuab microscope. Lub teeb nrog lub wavelength luv txaus kom txaus siab rau fluorophore yog illuminated ntawm tus qauv. Nrog kev pab los ntawm lub hom phiaj lens, lub teeb no yog tsom rau cov qauv. Cov excitation rays emitted los ntawm cov qauv yog ces tsom rau lub detector los ntawm lub hom phiaj lens. Qhov tseem ceeb ntawm lub teeb excitation yog kis los ntawm tus qauv khoom, thaum ib feem ntawm nws yog reflected rov qab thiab mus txog lub detector.
Fluorescence microscope Cheebtsam
1. Fluorophore
Ib qho fluorophore yog ib qho sib xyaw ua ke uas tawm lub teeb thaum zoo siab los ntawm electromagnetic los yog lub teeb nthwv dej. Nws tseem hu ua fluorescent dye. Fluorophores yeej ib txwm muaj kev sib xyaw ua ke ntawm cov tshuaj tsw qab lossis cov phiaj xwm / cyclic molecules nrog ntau yam π bonds.
2. Lub teeb qhov chaw
Lub teeb ci yog ib qho tseem ceeb tshaj plaws ntawm fluorescence microscope. Ntawm no, cov teeb pom kev zoo tshaj plaws siv rau kev ua kom muaj zog muaj xws li lasers, high-power LEDs, xenon arc teeb, mercury vapor teeb, thiab lwm yam.
3. Excitation lim
Excitation filters siv nyob rau hauv fluorescence microscopy feem ntau yog bandpass lim, uas tso cai rau cov teeb liab uas poob rau hauv ib qho bandwidth thaum tsis kam lees tag nrho lwm cov teeb liab. Lub wavelengths absorbed los ntawm fluorophore kis tau yooj yim, thaum lub reflected optical tawg yog thaiv.
4.Dichroic daim iav
Dichroic iav ua raws li cov ntxaij lim dej nyias. Lub hom phiaj tseem ceeb ntawm daim iav dichroic yog xaiv kom dhau lub teeb ntawm ib qho me me ntawm cov xim thaum xav txog tag nrho lwm cov xim. Nws tseem hu ua cov xim xim lim.
5. Tshem tawm lim
Lub emission lim yog lub lim bandpass. emission filters ua haujlwm zoo ib yam li cov lim dej excitation. Nws radiates lub teeb emitted los ntawm fluorophore thaum thaiv lub teeb excitation.
Qhov zoo ntawm Fluorescence Microscopy
1. Lub rhiab heev ntawm fluorescence microscopy yog siab heev, sib npaug ntawm kwv yees li 50 molecules ib cubic micrometer.
2. Nrog fluorescence microscopy, cov molecules sib txawv tuaj yeem dyed xim sib txawv, tso cai rau cov neeg siv duab ntau lub molecules ntawm ib yam khoom tib lub sijhawm.
3. Fluorescence microscopes muaj txiaj ntsig zoo dua li ib txwm siv tshuaj kho qhov muag.
4. tuaj yeem siv rau hauv vivo thiab hauv vitro duab.
5. Fluorescence microscopy tuaj yeem cais tus kheej cov proteins nrog kev ua haujlwm siab.
6. Fluorescence microscopy yog qhov zoo tshaj plaws rau kev kuaj pom thiab kawm txog cov cwj pwm tsis zoo uas pom los ntawm cov hlwb nyob.
7. Fluorescence microscopy muaj peev xwm ua kom muaj kev daws teeb meem siab thiab cov duab meej, yog li txhim kho kev tshawb fawb.
8. Nws muab cov khoom zoo ntawm cov khoom siv kho qhov muag nrog rau qhov pom ntawm fluorescence.
9. Fluorescence microscopy tuaj yeem siv los tsim 3D magnified dluab ntawm cov qauv uas tau kawm.
Disadvantages ntawm fluorescence microscopy
1. Fluorescent compounds los yog fluorophores uas muaj nyob rau hauv fluorescence microscopy yuav poob lawv lub peev xwm los emit lub teeb lub sij hawm thaum zoo siab vim yog ib tug txheej txheem hu ua photobleaching. Photobleaching feem ntau tshwm sim thaum lub fluorophore molecule raug puas tsuaj vim hluav taws xob excitation thaum lub sij hawm fluorescence.
2. Nws tsuas yog tso cai rau kev soj ntsuam thiab kawm txog qee cov qauv hauv cov qauv uas tau qhia thiab sau tias yog fluorescent.
3. Thaum cov khoom siv fluorescent yog irradiated los ntawm lub teeb, fluorescent molecules feem ntau tsim cov tshuaj, uas ntxiv rau phototoxicity. Cov nyhuv phototoxic no rhuav tshem cov khoom qub ntawm cov qauv pom. Lub phototoxicity ntawm luv-wavelength lub teeb hluav taws xob yog qhov siab.
4. Fluorescence microscopy tsis tuaj yeem muab cov ntaub ntawv hais txog lipid bilayer, tshwj xeeb tshaj yog rau SUVs.
5. Fluorescence microscopy tsis tuaj yeem ua tiav cov kev soj ntsuam uas muaj feem xyuam nrog cov cim bilayer lossis lamellarity ntawm cov vesicles.
6. Qhov sib ntxiv ntawm kev sojntsuam thiab dyes feem ntau cuam tshuam nrog cov khoom ntawm lipid vesicles, ua rau cov ntaub ntawv tsis raug txhais lus thiab yog li rhuav tshem qhov tseeb ntawm qhov kev sim.
7. Lub teeb-induced lipid peroxidation tej zaum yuav ua rau domain tsim.
Kev siv cov fluorescence microscopy
1. Fluorescence microscopy yog dav siv los kawm cov qauv sab hauv ntawm cov qauv ntawm qib microscopic nrog qhov zoo tshaj plaws.
2. Fluorescence microscopy yog qhov zoo tshaj plaws rau kev kawm cov qauv lom neeg nyob.
3. Hom kab mob no feem ntau nyiam thaum sau npe rau cov qauv hauv nruab nrab ntawm cov hlwb lossis cov kab mob.
4. Pab ntsuas lub cev lub cev ntawm cov hlwb lossis cov qauv kuaj pom.
