Optical Microscope Thickness Requirements rau iav Slides
Microscopic Dehydration thiab Sample Attachment: Vim yog rov centrifugation, cov hlwb ploj, yog li nws yog qhov zoo tshaj plaws los muab cov hlwb mus rau saum npoo ntawm lub iav swb los yog cov ntawv yas rau optical microscopy, kom lub hlwb dehydrated thiab qhuav ua ke nrog cov iav. swb los yog daim ntawv yas. Qhov yuav tsum tau ua rau lub thickness ntawm iav slides nyob rau hauv optical microscopy Yuav kom cov hlwb mus txuas rau cov iav slides los yog cov ntaub ntawv yas, nws yog ib qhov tsim nyog los ua ntej nteg ib txheej ntawm zaj duab xis rau iav slides los yog cov ntawv yas. Txoj kev tso zaj duab xis yog raws li nram no:
(1) Txoj kev yooj yim tshaj plaws rau optical microscopy yog nteg ib txheej nyias ntawm polyvinyl formal (formvar) zaj duab xis. So kom huv si rau hauv (0.5-1) feem pua polyvinyl formal chloroform tov. Tom qab huab cua volatilizes, ib zaj duab xis dawb tshwm rau ntawm cov slides. Co cov slides ob peb zaug hauv chloroform kom hloov cov yeeb yaj kiab. Nyias thiab npaj siv.
(2) Muab pov tseg 0.1 latitude poly-L-lysine (solve 1 mg poly-lysine hauv 1 ml lim dej thaum npaj) rau ntawm lub khob ntxuav, thiab muab tso rau hauv 450C lub thawv khoom tom qab cov tee dej nthuav tawm. . qhuav. Txoj kev no yog qhov zoo tshaj plaws vim tias poly-L-lysine tau them nyiaj zoo, tso cai rau ntau qhov chaw tsis zoo rau cov cell txuas.
(3) Ntxiv plasma thiab glycerol rau cov dej distilled ntawm qhov sib piv ntawm (30--40) feem pua thiab (3-5) feem pua raws li qhov yuav tsum tau muaj ntawm lub tshuab kho qhov muag rau qhov tuab ntawm cov swb los tsim cov glycerol protein. daws. Cov kua yog poob rau ntawm lub khob ntxuav, nthuav tawm thiab qhuav nyob rau hauv lub tshuab kho qhov muag. Cov yeeb yaj kiab tau muab tso rau hauv 2 feem pua glutaraldehyde los kho cov protein, ntxuav thiab qhuav rau tom qab siv. Ua ntej siv, cov plasma poob rau ntawm cov yeeb yaj kiab protein Ua kom nws "activated", yaug nrog tsis pub dhau 30 feeb, thiab nws tuaj yeem siv ncaj qha.
Cov cell ruaj khov thiab huv si yog tsim los rau hauv qhov tsim nyog ntawm kev tshem tawm ntawm tes nrog cov dej lim hauv qab lub tshuab ntsuas digital, thiab lub tshuab kho qhov muag tau muab tso rau ntawm lub iav swb lossis cov ntawv yas uas tau npog nrog daim nyias nyias, thiab muab tso rau hauv qhov chaw ntub, qis. qhov kub thiab txias (40C) ib puag ncig (30-120) feeb kom cia cov cell txuas mus rau daim nyias nyias. Siv cov tweezers zoo los khaws cov iav swb thiab co nws hauv cov dej huv huv ob chav distilled, thiab ntxuav cov hlwb uas tsis sib txuas nrog lub tshuab ntsuas qhov muag. Cov kev xav tau rau cov tuab ntawm cov slides hauv optical microscopes Polarizing microscopes yog ces muab tso rau hauv ib lub nkoj uas muaj 50 feem pua acetone (los yog ethanol). Cov txheej txheem lub cev qhuav dej yog muab cov slides rau hauv acetone los yog ethanol nrog nce concentration txhua 3 feeb, thiab kho qhov muag tsom iav los yog tsis tu ncua ua kom cov concentration ntawm cov dej dehydrating nyob rau hauv lub khob swb. Raws li tus sau qhov kev kuaj sim, kev ua haujlwm tom kawg yog yooj yim dua. Cov txheej txheem tshwj xeeb ntawm optical microscopy tuaj yeem nqus tau ib nrab ntawm tus neeg lub cev qhuav dej hauv lub thawv txhua 3 feeb, thiab tom qab ntawd ntxiv ib qho sib npaug ntawm tus neeg sawv cev lub cev qhuav dej nrog qhov concentration ntawm 100 feem pua. Tom qab 5-6 kev ua haujlwm, qhov siab ntawm tus neeg ua kom lub cev qhuav dej hauv lub tshuab hluav taws xob tus nqi nce siab tshaj 0.5 feem pua, thiab cov hlwb tau qhuav dej tiav. Tom qab cov ntsiab lus tseem ceeb kom qhuav, cov iav swb los yog cov ntawv yas yog glued rau cov qauv pier nrog cov nplaum nplaum rau cov hlau txheej. Kuj tseem muaj kev yooj yim rau kev txuas cov hlwb rau daim nyias nyias nteg los ntawm peb txoj kev. Optical microscope tuaj yeem dai cov hlwb uas tsis tshua muaj siab ntawm glutaraldehyde ntawm "activated" protein zaj duab xis. Optical microscope yuav tsum tau muab lub thickness ntawm cov iav swb rau 10 feeb, thiab cov iav swb los yog cov ntawv yas yog soaked nyob rau hauv 2 feem pua glutaraldehyde. Tom qab 30 feeb, yaug nrog lim ob chav distilled dej, thiab tom qab ntawd dehydrate, qhuav thiab hlau lub tsho tiv no raws li cov qauv saum toj no.
