Ukungcoliswa kwamasiko amaseli kalula kube inkinga ejwayeleke kakhulu kumalabhorethri e-cell culture, kwesinye isikhathi kubangele imiphumela emibi kakhulu.Ukungcola kwesiko lamaseli kungahlukaniswa ngezigaba ezimbili, ukungcola kwamakhemikhali njengokungcola okuphakathi, i-serum namanzi, ama-endotoxins, ama-plasticizers kanye nezinto zokuhlanza, kanye nokungcola kwezinto eziphilayo ezifana namagciwane, isikhunta, i-yeasts, amagciwane, i-mycoplasmas cross infection.Ingcoliswe abanye olayini beseli.Nakuba kungenakwenzeka ukuqeda ngokuphelele ukungcola, imvamisa nokuqina kwayo kungancishiswa ngokuqonda kahle umthombo wako nokulandela amasu amahle aseptic.
1.Lesi sigaba siveza izinhlobo eziyinhloko zokungcola kwebhayoloji:
Ukungcoliswa kwamagciwane
Ukungcoliswa kwesikhunta kanye negciwane
Ukutheleleka kwe-Mycoplasma
Ukungcola kwemvubelo
1.1 Ukungcola kwamagciwane
Amagciwane ayiqembu elikhulu lama-microorganisms aneseli elilodwa elitholakala yonke indawo.Ngokuvamile ama-microns ambalwa ububanzi futhi angaba nezimo ezihlukahlukene, ukusuka kuyimbulunga kuya kuma-rods kanye nama-spirals.Ngenxa yobuningi bawo, ubukhulu, kanye nezinga lokukhula ngokushesha, amabhaktheriya, kanye nemvubelo nesikhunta, yizona zinto ezingcolisa kakhulu eziphilayo esikweni lamaseli.
1.1.1 Ukutholwa Kokungcola Kwamagciwane
Ukungcoliswa kwamagciwane kutholakala kalula ngokuhlola isiko ngeso lengqondo ezinsukwini ezimbalwa ngemva kokuba litheleleke;
Amasiko angenwe yileli gciwane ngokuvamile abonakala eguqubele (okungukuthi, ama-turbid), ngezinye izikhathi anefilimu elincanyana phezulu.
Ukwehla okungazelelwe kwe-pH yendawo yesiko nakho kuvame ukuhlangana nakho.
Ngaphansi kwesibonakhulu esinamandla amancane, amagciwane avela njengamancane, ama-granules anyakazayo phakathi kwamaseli, futhi ukubuka ngaphansi kwesibonakhulu esinamandla amakhulu kungaxazulula izimo zebhaktheriya ngalinye.
1.2 Ukungcoliswa kwesikhunta kanye negciwane
1.2.1 Ukungcola Kwesikhunta
Isikhunta ama-microorganisms e-eukaryotic ombuso wesikhunta akhula ngendlela yama-multicellular filaments ebizwa ngokuthi i-hyphae.Amanethiwekhi axhumayo ale micu yamangqamuzana amaningi aqukethe ama-nuclei afana nofuzo abizwa ngokuthi amakholoni noma i-mycelium.
Ngokufana nokungcoliswa kwemvubelo, i-pH yesiko ihlala izinzile phakathi nesigaba sokuqala sokungcola bese ikhula ngokushesha njengoba isiko litheleleka kakhulu futhi liguquguquke.Ngaphansi kwe-microscope, i-mycelium ivame ukuba ne-filamentous, ngezinye izikhathi njengamaqoqo aminyene ezinhlamvu.Izinhlamvu zesikhunta eziningi zingaphila endaweni enzima kakhulu futhi engenamoya phakathi nesigaba sazo sokulala futhi zisebenza kuphela lapho izimo zokukhula ezifanele kuhlangatshezwana nazo.
1.2.2 Ukutheleleka ngegciwane
Amagciwane ayizisetshenziswa ezithathelwanayo ezincane kakhulu ezithatha imishini yeseli eliphethe ukuze zikhiqize kabusha.Ubukhulu bawo obuncane kakhulu bubenza kube nzima ukubonwa emasikweni kanye nokukhipha kuma-reagents asetshenziswa kumalabhorethri e-cell culture.Njengoba amagciwane amaningi enezidingo eziqine kakhulu kubasingathi bawo, ngokuvamile awathinti kabi amasiko amaseli ezinhlobo zezilwane ngaphandle komsingathi.
Kodwa-ke, ukusetshenziswa kwamasiko amaseli atheleleke ngegciwane kungaba yingozi enkulu empilweni yabasebenzi baselabhorethri, ikakhulukazi uma amaseli omuntu noma ama-primate ekhuliswa elabhorethri.
Ukutheleleka ngegciwane kumasiko amaseli kungatholwa nge-electron microscopy, i-immunostaining ngesethi yamasosha omzimba, i-ELISA, noma i-PCR eneziqalo zegciwane ezifanele.
1.3 Ukungcoliswa kwe-Mycoplasma
Ama-Mycoplasma ama-bacterium alula angenazo izindonga zamangqamuzana, futhi kucatshangwa ukuthi yizinto ezincane kunazo zonke eziziphindaphindayo.Ngenxa yobukhulu bawo obuncane kakhulu (imvamisa engaphansi kwe-micron engu-1), i-mycoplasma ilukhuni ukuyibona ize ifinyelele ukuminyana okuphezulu kakhulu futhi ibangele amasiko amaseli ukuba awohloke;Kuze kube yileso sikhathi, ngokuvamile azikho izimpawu ezisobala zokutheleleka.
1.3.1 Ukutholwa kokungcola kwe-mycoplasma
Amanye ama-mycoplasma akhula kancane angase aphikelele emasikweni ngaphandle kokubangela ukufa kwamangqamuzana, kodwa aguqule ukuziphatha kanye nokugayeka kokudla kwamangqamuzana aphethe emasikweni.
Ukutheleleka kwe-mycoplasma okungapheli kungase kubonakale ngokuncishiswa kwezinga lokukhula kwamangqamuzana, ukunciphisa ukugcwala kwe-saturation kanye ne-agglutination kusiko lokumiswa.
Kodwa-ke, okuwukuphela kwendlela enokwethenjelwa yokuthola ukungcola kwe-mycoplasma ukuhlola isiko njalo kusetshenziswa amabala e-fluorescent (isb, i-Hoechst 33258), i-ELISA, i-PCR, i-immunostaining, i-autoradiography, noma ukuhlolwa kwe-microbial.
1.4 Ukungcola kwemvubelo
Imvubelo ama-eukaryote aneseli elilodwa lombuso wesikhunta, asukela ngosayizi ukusuka kuma-microns ambalwa (imvamisa) kuya kuma-microns angama-40 (akuvamile).
1.4.1Ukutholwa kokungcola kweyesti
Njengokungcoliswa kwamagciwane, amasiko angcoliswe imvubelo angaguqubala, ikakhulukazi uma ukungcola sekusezingeni eliphezulu.I-pH yamasiko angcoliswe imvubelo ishintsha kancane kakhulu kuze kube yilapho ukungcola kuba kubi kakhulu, lapho i-pH ivame ukukhuphuka.Ngaphansi kwe-microscope, imvubelo ibonakala njengezinhlayiya ze-ovoid ngayinye noma eziyindilinga futhi ingase ikhiqize izinhlayiya ezincane.
2.Ukutheleleka ngegciwane
Nakuba kungavamile njengokungcoliswa kwamagciwane, ukungcoliswa okubanzi kwemigqa yamaseli amaningi nge-HeLa kanye neminye imigqa yamaseli ekhula ngokushesha kuyinkinga echazwe ngokucacile enemiphumela emibi.Thola imigqa yamaseli kumabhange amaseli ahloniphekile, hlola njalo izici zemigqa yamaseli, futhi usebenzise amasu amahle we-aseptic.Le mikhuba izokusiza ukuthi ugweme ukungcoliswana.Izigxivizo zeminwe ze-DNA, i-karyotyping kanye ne-isotyping kungaqinisekisa ukuthi kukhona ukungcoliswa okuphambene nesiko lamaseli akho.
Nakuba kungavamile njengokungcoliswa kwamagciwane, ukungcoliswa okubanzi kwemigqa yamaseli amaningi nge-HeLa kanye neminye imigqa yamaseli ekhula ngokushesha kuyinkinga echazwe ngokucacile enemiphumela emibi.Thola imigqa yamaseli kumabhange amaseli ahloniphekile, hlola njalo izici zemigqa yamaseli, futhi usebenzise amasu amahle we-aseptic.Le mikhuba izokusiza ukuthi ugweme ukungcoliswana.Izigxivizo zeminwe ze-DNA, i-karyotyping kanye ne-isotyping kungaqinisekisa ukuthi kukhona ukungcoliswa okuphambene nesiko lamaseli akho.
Isikhathi sokuthumela: Feb-01-2023