Rini mai sauƙi da shuɗin methylene da rinin Gram: abin da kowace hanya za ta iya tabbatarwa

Wani ɗalibi ya shirya samfurin da aka shimfiɗa a kan gilashin dubawa daga al'adar da aka yi wa lakabin E. coli, ya lulluɓe shi da shuɗin methylene na minti ɗaya, ya kurkure, sannan ya duba ta ruwan tabarau na amfani da mai. Ƙwayoyi shuɗi masu siffar sanda sun cika filin dubawa. Rahoton ɗakin gwaje-gwaje ya ce: "An gano ƙwayoyi masu siffar sanda na Gram-korau a matsayin E. coli." Abin da aka lura da shi daidai ne. Amma wannan gilashin dubawa bai ba da hujjar wannan ƙarshe ba.

Kuskuren yana cikin fahimta, ba cikin dabarar aiwatarwa ba. Ɗalibin ya ɗauki rini mai sauƙi kamar gwaji ne na bambancewa. Rini mai sauƙi da rinin Gram suna amsa tambayoyi daban-daban domin sun dogara da ƙa'idodin rini daban-daban. Ɗaya yana tambaya: wace siffa ce ake gani. Ɗayan yana tambaya: yaya bangon ƙwayar halitta yake nuna hali idan aka gwada shi. Idan aka rikita biyun, gilashin dubawa da aka yi wa rini daidai zai haifar da rahoton da ba shi da isasshiyar hujja.

Wannan kwatancen yana ware hanyoyin biyu, yana nuna abin da kowane sakamako yake ba ka damar faɗa, kuma yana amsa tambayoyi biyu da ɗalibai suka fi yi: ko za a iya amfani da shuɗin methylene a rinin Gram, da abin da rinin E. coli da shuɗin methylene yake nunawa a zahiri.

Amsa a taƙaice

Rini mai sauƙi da shuɗin methylene yana ba kowace ƙwayar halitta da za ta karɓi rini shuɗi iri ɗaya, don haka yana bayyana siffa (sanduna, ƙwallaye, karkace) da jeri (ɗaiɗaiku, bibbiyu, sarƙoƙi), amma ba ya nuna rukunin Gram. Rinin Gram yana ƙara iodine, matakin cire launi, da rini na biyu, don haka yana rarraba bakteriya zuwa Gram-tabbatacce (shunayya) da Gram-korau (hoda ko ja) bisa yadda suturar ƙwayoyinsu take riƙe rini. Sanduna shuɗi bayan amfani da shuɗin methylene suna nufin "akwai ƙwayoyin halitta masu siffar sanda." Sanduna masu launin hoda bayan cikakken jerin matakan Gram suna nufin "akwai ƙwayoyi masu siffar sanda na Gram-korau." Furuci na biyu ya fi ƙarfi domin hanyar ta aiwatar da ƙarin gwaji.

Don ƙarin bayani game da rukunin hanyoyin rinin bakteriya da rini guda ɗaya da wannan labarin yake ciki, duba kundin hotunan rini don dubawa ta madubin ƙara girma.

Rini mai sauƙi: rini ɗaya, tambaya ɗaya

Shuɗin methylene rini ne na asali mai cajin tabbatacce (cationic). Ion ɗinsa mai launi yana ɗauke da cajin tabbatacce kuma yana ɗaurewa da sassan ƙwayar halitta masu cajin korau, ciki har da acid ɗin nucleic da rukunin acid a saman ƙwayar, don haka ƙwayoyin suna bayyana shuɗi a kan bango mai haske. OpenStax ya bayyana wannan ƙa'idar rini kai tsaye ga ƙwayoyin halitta a babinsa na rini. Ion ɗin rinin mai cajin tabbatacce shi ne ƙaramin ion na phenothiazine mai faɗin siffa wanda aka yi bayani dalla-dalla a tushen bayanin sinadaran shuɗin methylene; ion ɗin chloride mai cajin kishiya ba shi ne yake ba da launin ba.

Ƙa'idar koyarwa da aka saba da ita tana da sauƙi da gangan: shimfiɗa samfurin siriri a kan gilashin dubawa, bar shi ya bushe a iska, tabbatar da mannuwarsa da zafi, lulluɓe shi da kusan 1% na shuɗin methylene da aka narkar a ruwa na kimanin minti ɗaya, kurkure a hankali da ruwa, tsotse ruwan har ya bushe, sannan a duba da ruwan tabarau da ake nutsawa a mai. Ƙa'idar koyarwa ta LibreTexts ta bayar da wannan tsarin na 1% na minti ɗaya daidai, tare da lura cewa wasu ɗakunan gwaje-gwaje suna tsawaita lokacin. Babu sinadarin tabbatar da rini, babu mai cire launi, babu rini na biyu. Wannan rashin ƙarin matakai shi ne muhimmin batu. Da rini ɗaya kawai kuma babu cire shi daga wasu ƙwayoyi kaɗai, kowace ƙwayar da ta ɗaure rinin za ta kasance da launi iri ɗaya.

Da E. coli, abin da ake sa ran gani a filin dubawa shi ne ƙwayoyi shuɗi masu siffar sanda (bacilli), ɗaiɗaiku ko bibbiyu ko a ƙananan rukuni. Shuɗin yana fitowa daga rinin. Siffar sanda tana fitowa daga ƙwayar halittar. Amma haɗuwar waɗannan ba ta tantance ƙwayar halittar, domin nau'o'i da yawa suna da siffar sanda kuma sanduna da yawa suna zama shuɗi a rini mai sauƙi. ATCC ya bayyana wannan iyaka ta nuna siffa kawai a jagoransa na shuka ƙwayoyin halitta. Rahoton da ya dace shi ne: "an ga bacilli shuɗi." Duk wani bayani da ya wuce siffa da jeri yana buƙatar ƙarin hujja.

Rinin Gram: sinadaran gwaji huɗu, gwajin hali ɗaya

Rinin Gram rini ne na bambancewa. Yana bi da dukkan ƙwayoyin halitta da jerin matakai iri ɗaya, sannan yana nuna waɗanne ƙwayoyi suka riƙe rini na farko da waɗanne suka rasa shi. Ƙa'idodin rinin Gram na American Society for Microbiology sun bayyana daidaitaccen jerin matakai huɗu: crystal violet (rini na farko), iodine na Gram (sinadarin tabbatar da rini), barasa ko cakudar acetone da barasa (mai cire launi), sannan safranin (rini na biyu).

Tsarin koyarwa da aka saba da shi yana bayyana bambancin ta ginin suturar ƙwayar halitta. Ƙwayoyin Gram-tabbatacce suna da bangon peptidoglycan mai kauri wanda yake rasa ruwa kuma ya riƙe haɗaɗɗen sinadarin crystal violet da iodine yayin wankewa da barasa. Ƙwayoyin Gram-korau suna da siririn shimfiɗar peptidoglycan da kuma membrane na waje mai yawan lipid wanda mai cire launi yake tarwatsawa, don haka haɗaɗɗen sinadarin yana fita yayin wankewa sannan ƙwayoyin su karɓi safranin. Bitar rinin Gram ta NCBI Bookshelf ta bayar da wannan bayanin bangon ƙwayar halitta. Ƙwararru suna ƙara da cewa cikakken tsarin hulɗar zahiri da sinadarai ya fi kaurin bango kaɗai rikitarwa, amma wajen fassara sakamako a ɗakin gwaje-gwaje, ƙa'idar halin tana nan: shunayya yana nufin an riƙe rini, hoda yana nufin an rasa shi sannan aka karɓi rini na biyu.

Zane-zanen misali. Zane ne don kwatanta hanyoyi, ba hotunan kamara da aka ɗauka ta madubin ƙara girma ba. An sauƙaƙa launukan don koyar da ƙa'idar: launi ɗaya a rini mai sauƙi, launuka biyu bayan cire launi a tsarin Gram.

Ko za a iya amfani da shuɗin methylene a rinin Gram

Ana iya fahimtar wannan tambayar ta hanyoyi biyu, kuma suna buƙatar amsoshi daban-daban.

Fahimta ta farko: ko lulluɓe samfurin da shuɗin methylene sau ɗaya zai iya maye gurbin dukkan jerin matakan Gram. A'a. Ba tare da iodine, mai cire launi, da rini na biyu ba, babu matakin bambancewa, don haka ba a gwada wani rukunin Gram ba. Filin shuɗi bayan amfani da shuɗin methylene shi kaɗai sakamakon rini mai sauƙi ne, ko da kwalbar tana kusa da sinadaran gwajin Gram. Kada a rubuta shi a matsayin sakamakon Gram.

Fahimta ta biyu: ko shuɗin methylene zai iya maye gurbin safranin a matsayin rini na biyu a mataki na 4. Ta fuskar aiki, zai iya ba ƙwayoyin da aka cire wa launi sabon launi, kuma atisayen littafin koyarwa yana hasashen ƙwayoyin Gram-tabbatacce masu launin shunayya da ƙwayoyin Gram-korau masu launin shuɗi bayan irin wannan sauyin. Wannan tunanin yana da tushe domin ƙwayoyin Gram-korau da aka cire wa launi yadda ya kamata kusan ba su da launi kuma za su karɓi wani rini mai cajin tabbatacce. Amma wannan ba ƙa'idar da aka saba da ita ba ce. Ƙa'idodin ASM sun tanadi safranin, tare da wasu madadin da aka amince da su kamar basic fuchsin a wasu nau'o'in tsarin, ba shuɗin methylene ba. Haka kuma akwai wata matsala a aikace: bambance shuɗi da shunayya ya fi bambance hoda da shunayya wahala, musamman ga masu farawa da kuma idan fitilun madubin ƙara girma sun raunana. Don aikin karatu da duk wani sakamako mai muhimmanci, yi amfani da rini na biyu da ƙa'idarka ta ambata, wanda yawanci safranin ne, kuma ka ɗauki amfani da shuɗin methylene a matsayin rini na biyu a matsayin wani tsarin nuna misali maimakon daidaitacciyar hanya mai matsayi iri ɗaya.

An tattauna tambayar da ke da alaƙa da wannan, wato yadda shuɗin methylene yake idan aka kwatanta shi da sauran zaɓuɓɓukan rini guda ɗaya, a kwatancen hanyoyin rini mai sauƙi na bakteriya.

Abin da rinin E. coli da shuɗin methylene yake tabbatarwa, da abin da ba ya tabbatarwa

E. coli ƙwaya ce mai siffar sanda ta Gram-korau, amma wannan jimlar tana haɗa bayanai daga hanyoyi da dama: siffa daga dubawa ta madubin ƙara girma, amsar Gram daga rinin Gram, tantance nau'i daga shuka ƙwayar tare da gwajin sinadaran halitta ko na ƙwayoyin molekul. Samfurin da aka yi wa rini da shuɗin methylene yana ba da ɓangaren farko kawai. Yana tabbatar da cewa akwai ƙwayoyin halitta masu siffar sanda a samfurin da aka yi wa rini, da yadda suke jere. Ba ya tabbatar da amsar Gram, domin ba a gudanar da gwajin cire launi ba. Ba ya tantance nau'in ƙwayar, domin sandunan nau'o'i da yawa suna kama da juna a wannan matakin iya rarrabe cikakkun bayanai.

A nan ne rahotanni suke yin kuskure ta hanyar da ake iya hasashe. Marubucin yana shigar da ilimin da yake da shi tun farko ("ana sa ran wannan al'adar ta kasance E. coli, kuma E. coli Gram-korau ne") cikin layin abin da aka lura da shi, don haka gilashin dubawa ya zama kamar yana tabbatar da abin da a zahiri aka ɗauka ne kawai. Rahoto mafi tsabta yana ware matakan bayanin: "Rini mai sauƙi da shuɗin methylene: an ga bacilli shuɗi; siffa kawai, ba a gwada rukunin Gram ba." Sannan, idan an yi rinin Gram daidai tare da samfuran sarrafa ingancin gwaji: "Rinin Gram: sanduna masu launin hoda, sun dace da Gram-korau." Tantance nau'in ƙwayar yana zuwa daga baya, daga tsarin tantancewa, ba daga launi kaɗai ba.

Jagorar fassara sakamako a aikace

Yi amfani da wannan ƙa'idar yanke shawara a ɗakin gwaje-gwaje. Idan dukkan ƙwayoyin suna da launi ɗaya kuma ƙa'idar ta yi amfani da rini ɗaya ba tare da mai cire launi ba, ka yi rini mai sauƙi. Ka bayar da rahoton siffa, ƙiyasin girma, da jeri. Ka tsaya a nan. Idan ƙa'idar ta yi amfani da crystal violet, iodine, mai cire launi, da rini na biyu tare da samfuran sarrafa ingancin gwaji da aka san suna da amsar tabbatacce da korau, ka yi rinin Gram. Ka bayar da rahoton launi tare da siffa: ƙwayoyi masu siffar ƙwallo na shunayya a cikin sarƙoƙi, sanduna masu launin hoda, da sauransu.

Kuskure uku ne suke haifar da mafi yawan ruɗani. Na farko, ɗaukar sanduna shuɗi a matsayin tabbacin E. coli fiye da abin da sakamakon yake nunawa. Na biyu, rubuta bayanin gilashin dubawa na shuɗin methylene ƙarƙashin taken "Rinin Gram" a littafin rubutu, wanda masu karantawa daga baya za su fassara ba daidai ba. Na uku, sauya rini na biyu ba tare da kulawa ba sannan a kwatanta launuka tsakanin ɗakunan gwaje-gwaje kamar an yi amfani da launuka iri ɗaya. Maganin kowane kuskure ɗaya ne: ambaci hanyar da aka aiwatar a zahiri, kuma ka faɗi abin da wannan hanyar take gwadawa kawai.

Rini mai sauƙi yana da muhimmanci ne musamman domin abin da yake tabbatarwa yana da iyaka. Yana da sauri, ƙananan kurakurai ba sa saurin ɓata shi, kuma ya isa idan tambayar ta shafi siffa: ko akwai wani abu, kuma wace siffa yake da ita. Idan tambayar ta shafi rarrabawa, yi gwajin bambancewa. Gilashin dubawa mai shuɗi yana gaya maka yadda ƙwayoyin suke bayyana. Gilashin mai launin shunayya ko hoda yana gaya maka yadda bangonsu yake nuna hali. Babu ɗaya daga cikin gilashin biyu, shi kaɗai, da yake gaya maka nau'in ƙwayar.