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Labarai

Tambayoyin Al'adun Salula da ake yawan yi

1. Idan na karɓi bututu na sel daskararre, zan iya saka shi kai tsaye cikin ruwa nitrogen don ajiya?

A lokuta da yawa, ƙwayoyin da ake jigilar su akan busassun kankara (-80°C) ana iya mayar da su cikin ruwa nitrogen sannan a narke da sauri daga baya.Duk da haka, ana iya rage yiwuwar tantanin halitta bayan irin wannan magani.Ga wasu layukan tantanin halitta, wannan na iya sa farfadowar tantanin halitta ya fi wahala.Ana tsammanin wannan al'amari ya faru ne saboda canjin tsarin lu'ulu'u na kankara a cikin sel sakamakon canjin yanayin zafi.Don haka ana ba da shawarar cewa sel su narke kuma a al'ada su da wuri-wuri bayan an karɓa.Rage lokacin ajiya a -80 ° C.Ana amfani da wannan zafin jiki ne kawai don sufuri.

Al'adun Kwayoyin Tambayoyi akai-akai 1
Al'adun Kwayoyin Tambayoyi akai-akai 2
Al'adun Kwayoyin Tambayoyi akai-akai 4
Al'adun Kwayoyin Tambayoyi akai-akai 3

2. Wadanne matakan tsaro ya kamata a ɗauka yayin cire sel daga nitrogen na ruwa don murmurewa?

Kwayoyin cryotubes a cikin ruwa na nitrogen waɗanda ba a rufe su gaba ɗaya kuma suna da ruwa na nitrogen da ke zubowa a cikin su na iya haifar da fashewa idan yanayin cryotube ya tashi sosai lokacin narke.Don haka ana ba da shawarar cewa a sanya tabarau da safar hannu masu kariya lokacin cire sel daga nitrogen mai ruwa.Don farfaɗowa, bututun mai daskarewa yakamata a girgiza gabaɗaya a cikin ruwan wanka na 37°C don narke gaba ɗaya maganin daskarewa cikin mintuna 1-2.Bayan haka, shafa waje na bututu tare da shafan barasa, sa'an nan kuma ɗauka a cikin tebur mai tsabta mai tsabta kuma canja wurin sel zuwa bututun centrifuge tare da 10 ml na al'ada da aka kara, centrifuge a 1000 rpm na minti 5-10, jefar da mai girma, ƙara adadin matsakaicin al'ada da ya dace kuma a saka flask ɗin al'ada kuma a saka a cikin incubator 5% CO2.

3. Me ya sa za a adana sel a cikin lokacin tururi na tankin nitrogen maimakon a cikin lokacin ruwa?

Kwayoyin da aka adana a lokacin iskar iskar gas na nitrogen ruwa suna da yuwuwar sake farfado da su.Ganin cewa a cikin lokacin ruwa na nitrogen na ruwa, idan ba a rufe bututun lyophilisation da kyau ba ko kuma suna da ɗigogi, hulɗa kai tsaye tsakanin sel da nitrogen ruwa na iya yin lahani ga yuwuwar sel bayan narke.

4. Don ƙwayoyin dakatarwa, ta yaya zan canza matsakaicin al'ada?

Za a iya yin al'adar sel dakatarwa ta hanyar ƙara sabon matsakaici (idan sarari ya ba da izini) ko ta hanyar raba sel daga tsohuwar matsakaici ta hanyar centrifugation (100 xg na mintuna 5) sannan a sake dawo da sel da aka haɗe a cikin sabon matsakaici.Koyaya, don yawancin layin salula na dakatarwa, ƙara matsakaici kawai shine mafi kyawun hanya.Ko ta yaya, matsakaicin yana buƙatar sabuntawa kafin sel su kai ga mafi girman jikewa.Yawan jikewa na sel ya bambanta tsakanin 3 x 10 5 da 2 x 10 6 dangane da layin salula da yanayin al'ada (hutawa ko motsawa, matakan oxygenation, da sauransu).Dole ne a narkar da ƙwayoyin sel zuwa ƙananan ƙwayar tantanin halitta don ba da damar isassun farfadowa na gina jiki don kiyaye sel suna girma logarithmically.Idan an canza matsakaicin kawai ba tare da rage yawan tantanin halitta ba, ƙwayoyin za su ƙare da sauri kuma su mutu.Idan an narkar da ƙwayoyin sel a ƙasa da ƙaramin adadinsu, za su shiga wani lokaci mai tsawo kuma su yi girma a hankali ko kuma za su mutu.Kowane layin salula na dakatarwa yana da mabambanta madaidaicin saturation da tazarar wucewa, don haka ƙidayar tantanin halitta ta yau da kullun ita ce hanyar saka idanu kan layukan salula*.

5. Menene shawarar CO2 matakin don al'adun tantanin halitta?

Kodayake matakan CO2 a cikin tsarin al'adun tantanin halitta sun fito daga 0.03% zuwa 40% (yawanci a kusa da 0.03% CO2 a cikin yanayi), yana da yawa don samun CO2 da aka kara a cikin iska ko CO2 maida hankali na 5% zuwa 10%.Yana da mahimmanci don daidaita daidaituwar sodium bicarbonate a cikin matsakaici don daidaitawa tare da matakin CO2 a cikin lokacin gas.Kwayoyin suna samar da CO2 kuma suna buƙatar ƙaramin adadin carbonic acid don girma da rayuwa.Idan ba a ƙara CO2 ba kuma ƙwayoyin suna haɓaka, ana iya amfani da 4 mM (0.34 g/L) na anhydrous sodium bicarbonate.Duk da haka, ya kamata a danne murfin flask ɗin al'ada a wannan lokacin.Idan tsarin al'ada yana buƙatar 5% ko 10% CO2, yi amfani da 23.5 mM (1.97 g / L) ko 47 mM (3.95 g / L) sodium bicarbonate a 37 ° C, bi da bi, tare da pH na farko na kusan 7.6.A karkashin waɗannan sharuɗɗan, yakamata a bar filashin ɗin ba a rufe ko a yi amfani da tasa Petri don kula da daidaiton iskar gas.

6. Me yasa wasu sel ke buƙatar sodium pyruvate?Nawa sodium pyruvate zan ƙara zuwa matsakaici?

Pyruvate shine metabolite na kwayoyin acid a cikin hanyar glycolytic * wanda ke shiga cikin sauri ya fita daga tantanin halitta.Sabili da haka, ƙari na sodium pyruvate zuwa matsakaici yana samar da tushen makamashi da kuma tushen carbon don anabolism, yana taimakawa wajen kula da wasu takamaiman ƙwayoyin cuta, yana taimakawa tare da cloning cell ko kuma ana buƙata lokacin da aka rage yawan ƙwayar jini a cikin matsakaici.Sodium pyruvate kuma yana taimakawa wajen rage cytotoxicity da ke haifar da fluorescence.Sodium pyruvate yawanci ana ƙara shi a matakin ƙarshe na 1 mM.Samfurin kasuwancin sodium pyruvate mafita yawanci 100 mM ajiya bayani (100X).

Fassara da www.DeepL.com/Translator (sigar kyauta).


Lokacin aikawa: Juni-21-2022