Biomicroscopy nrog rau qhov ntim ntawm cov ntaub so ntswg
Biological Microscopy Txog tam sim no, kev kho mob khaub thuas, khov ultrathin seem thiab khov-drying yog cov txheej txheem niaj hnub rau cov ntaub so ntswg thiab cell x-ray microsections. Cov ntsiab lus ntawm txoj kev no tau piav qhia raws li hauv qab no:
Rau cov tshuab kuaj kab mob lom neeg nrog lub condenser, lub condenser tuaj yeem txav mus los ua kom lub teeb ci nruab nrab, thiab qhov aperture ntawm lub teeb hloov pauv tuaj yeem hloov pauv kom ua tiav qhov nruab nrab 9b ci ntsa iab. Yog tias lub teeb ci tshav ntuj, lub condenser tuaj yeem tsa tau tsim nyog, thiab qhov aperture ntawm qhov sib txawv ntawm lub teeb tuaj yeem nthuav dav kom tsim nyog. Yog tias lub teeb muaj zog heev, lub condenser lo ntsiab muag tuaj yeem txo qis kom tsim nyog, thiab qhov aperture ntawm lub teeb sib tshuam tuaj yeem raug txo kom tsim nyog. Yog hais tias koj tseem xav tias dazzling nyob rau hauv cov ntaub ntawv no, koj muaj peev xwm xaiv ib tug tsim nyog lim thiab muab tso rau hauv lub bracket nyob rau hauv lub condenser. Cov ntoo qhib no yuav tuaj yeem tau txais qhov ci ntsa iab uas txaus siab rau koj. Ntawm chav kawm, kho lub sab sauv thiab qis txoj hauj lwm ntawm condenser, kho qhov aperture loj ntawm qhov sib txawv ntawm qhov kev nyeem ntawv kho qhov muag thiab xaiv cov lim tsim nyog yuav tsum muaj qee lub sijhawm ntawm kev xyaum kom tau txais kev paub.
Ib qho teeb meem tseem ceeb heev hauv kev siv tshuaj tua kab mob yog tias nyob rau hauv tus txheej txheem ntawm kev kuaj thiab cais cov hlwb, tom qab khov-qhuav thiab resin embedding (FD), tom qab khov ultra-thin aggregates, thiab tom qab khov-qhuav, cov ntsiab lus ntawm 65 ntsiab lus nyob rau hauv txhua feem. yuav tsum ua tib zoo saib xyuas. Cov hlwb yuav raug tshuaj xyuas tsis tuaj yeem raug puas tsuaj. Vim tias X-ray microanalysis tsis tsuas yog cuam tshuam nrog ntau cov kauj ruam, tab sis kuj raug nqi ntau heev. Yog tias cov hlwb uas tau txheeb xyuas yog cov hlwb puas lossis cov hlwb tuag tom qab lub sijhawm ntev thiab kev ua haujlwm ntau, nws yog qhov zoo siab heev los txiav txim siab tsis ncaj ncees lawm. Piv txwv li, cardiomyocytes sib cais los ntawm kev kho mob collagenase muaj ob daim duab, ib qho yog tus pas nrig ntev, thiab lwm qhov yog puag ncig. Cov tom kawg yog cov hlwb tuag uas raug puas tsuaj thaum lub sijhawm sib cais ntawm tes.
Biomicroscopy Tus nqi thiab kev faib tawm ntawm electrolytes hauv ob hom cell no txawv heev. Na yog siab heev thiab K yog tsawg heev nyob rau hauv lub voj voog cardiomyocytes, thiab cov concentration ntawm ca hauv nematodes siab heev. Tom qab kuaj xyuas nrog lwm txoj kev tshuaj ntsuam xyuas, nws tau ua pov thawj tias qhov siab Na thiab qis K ntawm puag ncig lub hlwb thiab Ca siab hauv mitochondria yog vim kev puas tsuaj ntawm cov cell membrane thaum lub sijhawm sib cais ntawm tes. Txoj kev kho dej khov-txias ntawm cov hlwb thiab cov ntaub so ntswg feem ntau yog siv quenching thiab fixation ua ntej, thiab tom qab ntawd muab cia rau hauv cov kua nitrogen. Quenching fixation yog ib qho tseem ceeb heev rau cov nyhuv ntawm preservation. Cov cell nyob los yog cov ntaub so ntswg tshiab muaj dej ntau. Thaum quenching, qhov chaw ntawm cov hlwb los yog cov ntaub so ntswg uas nyob rau hauv kev sib cuag ncaj qha nrog crushed refrigerant (tshwj xeeb tshaj yog thaum quenching nrog kua nitrogen) feem ntau khov thiab tsau ua ntej, yog li tsim ib "plhaub", uas cuam tshuam lub hauv paus ntawm lub hlwb. crushed thiab txias-kho. Yog li ntawd, thaum ua X-line micro-chaw tsom xam, nws feem ntau pom tias muaj cov dej khov hauv nruab nrab ntawm cov hlwb loj. Txhawm rau tiv thaiv qhov no los ntawm qhov tshwm sim, ib yam khoom uas muaj cov ntsiab lus melting siab dua li cov kua nitrogen tab sis qis dua li ntawm 806c yog siv los ua cov tub yees tawg. Muaj ntau yam khoom zoo li no, tab sis tus nqi pheej yig tshaj plaws yog cov concentrated propane (qhov kub ntawm - 42.120c, melting point - 187.10c, molecular hnyav 44.1), thiab qhov txias txias yog qhov ceev tshaj plaws. Tab sis nws qhov tsis zoo yog tias nws yog flammable.
Lub tshuab siv tshuaj lom neeg tuaj yeem muab cov nqaij fiber ntau tso rau ntawm ib qho tshwj xeeb khib, yog li ntawd thaum cov nqaij fiber ntau contraction mus txog ib theem thiab yuav tsum tau kho, tam sim ntawd pib lub nozzle los tsuag kua propane rau cov nqaij fibers kom quench thiab kho nws. Tom qab ntawd cov nqaij fibers ua ke nrog cov thav duab raug muab tshem tawm thiab muab tso rau hauv cov kua nitrogen. Yog tias kho cov qe ntshav los yog cov hlwb cais, ua ntej mloog cov hlwb los ntawm centrifugation ntawm qis ceev, hloov cov hlwb mus rau cov nyiaj vial nrog thermal conductivity zoo, tso lub vial rau hauv cov kua propane, thiab khov rau fixation. Thaum kho cov nas pancreas nyob rau hauv Hall Laboratory, ob lub steel blocks tau pre-txias nrog kua helium (los yog kua nitrogen), thiab ob lub tooj liab blocks raug clamped nrog forceps thiab muab tso rau pem hauv ntej thiab sab nraum qab ntawm lub txiav kom quench thiab kho. pancreas. Cov ntaub so ntswg los yog cov hlwb tuaj yeem khaws cia hauv cov kua nitrogen rau lub sijhawm ntev tom qab quenching thiab kho.
Biological Microscopy Ntxiv rau qhov kev hwm frozen ultrathin seem txoj kev, ntawm no yog lwm cov txheej txheem tso nyiaj siv los ntawm cov kws tshawb fawb. Ib qho tshuaj ntxiv los tsim cov sediment nrog rau cov khoom siv los soj ntsuam kom tsis txhob muaj kev cuam tshuam ua ntej kev tshuaj ntsuam, thiab cov sediment tau txheeb xyuas. Piv txwv li, tib neeg feem ntau siv oxalate thiab pyroantimonate tso ca hauv cov leeg nqaij. Cov qub tsis rhiab txaus rau qhov qis concentration ntawm Ca hauv cytoplasm; pyroantimonate yog rhiab heev thiab muaj peev xwm tsim electron-dense deposits nrog dawb Ca nyob rau hauv lub hlwb, tab sis pyroantimonate tsis sib xws nrog sodium, manganese, barium, hlau deposits kuj tsim thiab tsis tshua muaj. Cov txheej txheem kev npaj cov qauv zoo ib yam li kev npaj cov khoom sib kis ntawm electron microscope ultrathin seem cov qauv, qhov txawv yog tias 3 feem pua poov tshuaj pyroantimonate (phosphate buffer saline, pH 7.6) tau kho rau 6 teev ua ntej kho, thiab ntawm cov leeg nqaij ultrathin, Cov deposits dub tau pom nyob rau hauv nruab nrab ntawm bands A thiab 2 ntawm txhua sarcomere, thiab cov seem tsis muaj xim tau siv rau X-ray microanalysis. Diaminobenzidine tetrahydrochloride (DAB) tau siv los ua kom cov khoom muaj heme thiab lwm yam. Kev sib xyaw ua ke ntawm histochemical nag lossis daus cov tshuaj tiv thaiv thiab x-ray micro-differentiation choj tuaj yeem raug txiav txim siab hauv cov chaw soj nstuam uas tsis muaj cov txheej txheem khov kho ultrathin. Semi-quantitative tuaj yeem ua tau raws li qhov siab tshaj plaws ntawm cov ntsiab lus los txheeb xyuas
