Indlela yokukrazula-Yenza iCacterial Culture (Lyophilization)

Ukuphilisa i-Lyophilization, okanye ukumisa umisa, kuyinkqubo emfutshane yebhubhoratri

Ukumisa ukutshaya, okubizwa ngokuba yi- lyophilization okanye i-cryodesiccation, yinkqubo yokususa amanzi kwimveliso emva kokuqhwala (kwaye ibekwe phantsi kwe-vacuum) ukuvumela i-ice ukuba ishintshe ukusuka kwindawo eqinile ibe ngumphunga, ngaphandle kokuhamba kwisigaba samanzi. Iqhwa (okanye ezinye i-solvents eziqhotyoshelweyo) zisuswe kwimveliso ngokusebenzisa inkqubo ye-sublimation kunye neamolekyu zamanzi ezibophekileyo zisuswa kwinkqubo yokuchithwa.

Iisiseko zeLyophilization

Enye yezona ndlela ezintle zokugcina ibhakteria, i-fungal, imvubelo okanye enye inkcubeko ye-microorganism kwixesha elide lokusebenzisa i-process of frozen-drying. Le nkqubo emfutshane yebhoratri ingaqhutyelwa nayo nayiphi na i-dryer-dryer ekhoyo yokuthengisa ifumaneka kuyo eya kulondoloza ukuqokelela kwenkcubeko. Ngenxa yokuba i-lyophilization yindlela eyinkimbinkimbi kakhulu kunye neyabizileyo yokumisa, le nkqubo ivame ukukhawulelwa kwizinto ezinqabileyo, ezinokushisa ukushisa. Izinto ezingonakaliswe ngokuqhwaba zinokuthi zenziwe lyophilized ukwenzela ukuba ukugcinwa kwefriji akudingekile.

Le nkqubo ingathatha nje ngeeyure ezintathu, kwaye zide iiyure ezingama-24, kungabandakanyi ixesha lokukhula kwenkcubeko.

Inkqubo yeNyathelo-ngeNyathelo yeLyophilization

1. Okokuqala, khula inkcubeko yobusuku, okanye ingca, ye-microorganism kwi-LB okanye enye i-agar iplate.

2. Emva koko, lungiselela izibilini ze-crimp-cap-clock nge-autoclaving ngaphambi kwexesha, kunye neefowuni (izitofu zeerubha) zifakwe ngokukhululeka phezulu. Beka iileta zamaphepha eziprintwayo kunye nenkcazo yenkcubeko ngaphakathi kwiibhubhu ngaphambi kokuba i-autoclaving. Ngaphandle koko, sebenzisa iibhubhu ezinama-caps ezenzelwe ukubola .

1. Yongeza u-4 ml we-llophization buffer kwi-plate. Ukuba kuyimfuneko, iiseli zinqunyanyiswe usebenzisa intonga yeglasi engasinyanisi.

2. Ngokukhawuleza udlulisele inkcubeko yokumiswa kwinqwelo ezinamanzi. Yongeza malunga ne-1.5 mL nge-vial. Gcoba ngekhuni yerabha.

3. Ukukhulula ukunyuswa kwenkcubeko ngaphakathi kwezibilini ngokufaka iifom kwi-degrees Celsius freezer.

1. Emva kokuba iinkcubeko ziqingqiwe, lungiselela umququzeleli ngokuziphendulela kwaye uvumele ixesha elifanelekileyo lokushisa kunye neemeko zokuphucula. Yenza oku ngokuya kumyalelo womenzi we-brand ye-frozen-dryer oyisebenzisayo.

2. Ngenyameko, kunye ne-aseptically, faka iinqwelo ze-vial ngokuzenzekelayo ngaphezulu kweengqayi, ukwenzela ukuba umswakama ungaphunyuka ngexesha lokumisa i-frozen, kwaye ubeke iingqayi kwiklasi elomileyo. Faka isicelo sokukhupha egumbini ngokwemigaqo yomenzi.

3. Vumela ixesha lekcubeko ukuba litshise ngokupheleleyo (omile). Oku kungathabatha kwiiyure ezimbalwa ukuya kobusuku ngokuxhomekeke kumthamo wesampula ngasinye kwaye zingaphi iisampuli onayo.

4. Susa iisampuli kwikamelo lokumisa i-frozen ngokwemigaqo yomenzi kwaye unyule ngokukhawuleza izibhambathiso kunye neefowuni zeerubha, ngoko ke ukukhupha.

5. Gcina iqoqo lenkcubeko lyophilized kwiqondo lokushisa.

Imveliso oya kuyidinga

• I-Freeze-dryer
• I-nutrient okanye enye i-agar ipates (kunye ne-incubator ukukhula kwenkcubeko)
• Intonga yeglasi
• I-buyophilization buffer
• Izikhuni eziphezulu ze-crimp kunye ne-raber stoppers (kunye nomntu okhuselayo wokusebenzisa i-caps)
• IFreezer

<sub>Imbekiselo:

Scientific SP. Imigaqo-siseko yeFreeze Ukucwina. http://www.spscientific.com/freeze-drying-lyophilization-basics/</sub>