New methylene blue no gusiga retikulocyte: irangi ritandukanye n'uburyo bukoreshwa ku turemangingo tukiri tuzima

Umunyeshuri yitegura kubara retikulocyte, agatumiza methylene blue, akayivanga n'amaraso, maze ntihagire hafi ikintu abona. Mugenzi we akoresha icupa ryanditseho new methylene blue, akabona ududodo tw'ubururu tugaragara neza imbere muri bimwe mu turemangingo dutukura tw'amaraso. Umunyeshuri wa mbere yibwira ko irangi ryo mu icupa rya kabiri ari rishya kurushaho cyangwa ko rifite ubukana bwinshi. Nta na kimwe muri ibyo ari cyo. Ririmo molekile itandukanye.

Uku kutumva neza ni ko gutera urujijo rwinshi muri uru rwego. Izina rituma umuntu atekereza ko new methylene blue ari methylene blue yanonosowe. Imiterere yabyo ya shimi igaragaza ibinyuranye n'ibyo. New methylene blue N ni irangi ryihariye ryo mu muryango wa phenothiazine, rifite nomero yaryo muri Colour Index, imiterere yaryo y'amatsinda yometse kuri molekile, n'uruhare rwaryo rugenwa n'ibipimo ngenderwaho mu bumenyi bw'amaraso. Iyo iryo tandukaniro rimaze kumvikana, ibindi bigize ubu buryo biroroha: impamvu uturemangingo tugomba kuba tukiri tuzima igihe dusigwa, ibarwa nka retikulocyte, aho utubumbe twa Heinz duhuriye na byo, n'igihe brilliant cresyl blue ikoreshwa.

Igisubizo kigufi

New methylene blue N, inyongeramusaruro ikoreshwa mu bumenyi bw'amaraso, isanzwe imenyekana nka Basic Blue 24 mu makuru ya PubChem yerekeye new methylene blue, C.I. 52030, CAS 1934-16-3, ifite fomula C18H22ClN3S. Methylene blue isanzwe ni methylthioninium chloride, Basic Blue 9, mu makuru yayo ya PubChem, C.I. 52015, CAS 61-73-4, ifite fomula C16H18ClN3S. Mu miterere yayo, new methylene blue ifite amatsinda abiri ya methyl ku mpeta ya molekile n'amatsinda ya ethylamino ku mpande, aho methylene blue ifite amatsinda ya dimethylamino. Iryo tandukaniro rihindura uko zisiga uturemangingo. Inyandiko z'ubumenyi bw'amaraso zisobanura ko methylene blue isanzwe ari irangi ridakora neza cyane kuri retikulocyte, naho new methylene blue ikazitera ibara rikomeye kandi risa neza hose; iryo gereranya ryavunaguye mu nyandiko ya ScienceDirect yerekeye new methylene blue.

Yego, new methylene blue ni irangi rikoreshwa ku turemangingo tukiri tuzima, uburyo bwitwa supravital. Gusiga mu buryo bwa supravital bisobanura gusiga uturemangingo tukiri tuzima nyuma yo gukurwa mu mubiri, ariko mbere yo kudufatisha, nk'uko bisobanurwa mu isuzuma rya NCBI ry'amahame yo gusiga mu buryo bwa supravital. New methylene blue na brilliant cresyl blue byombi biri muri iki cyiciro, kandi igipimo ngenderwaho cya CLSI kuri retikulocyte kivuga mu buryo bweruye ko new methylene blue ari irangi rya supravital. Rimwe na rimwe abanyeshuri bashyira ubu buryo mu cyiciro cyo gusiga ibyasizwe ku kirahure kandi byamaze gufatishwa bakoresheje uburyo bwa Romanowsky. Iyo myumvire ntikora hano. Iyo ubanje gufatisha uturemangingo, ubu buryo ntibukora.

Ku basomyi bashaka gutandukanya amazina y'amarangi asa, methyl blue ugereranyije na methylene blue isobanura urundi rujijo runini hagati y'irangi rya triarylmethane rifite umuzigo w'amashanyarazi mubi n'umuryango wa phenothiazine ufite umuzigo mwiza. Incamake yo gusiga muri laboratwari ishyira uburyo bwa supravital mu rwego rumwe n'uburyo bukoresha ibyasizwe ku kirahure kandi byafatishejwe, naho igitabo kiyobora ku miterere y'uturemangingo tw'amaraso kigaragaza aho gusuzuma retikulocyte bihuriye no gutandukanya amoko atandukanye y'uturemangingo tw'amaraso.

Icyitonderwa kimwe ku rutonde rw'ibicuruzwa: hari ubundi bwoko bwa new methylene blue bugizwe n'umunyu w'ibiri urimo zinc chloride, bufite CAS 6586-05-6. Bityo, igice gisobanura uburyo bwakoreshejwe kigomba kuvuga ubwoko bw'umunyu n'uwawutanze, aho kuvuga izina risanzwe gusa.

Icyo gusiga mu buryo bwa supravital bikora koko

Retikulocyte ni akaremangingo gatukura k'amaraso kakiri gato, kaba kamaze gutakaza intima yako ariko kagifite ibisigazwa bya RNA ya ribosome. Mu cyitegererezo gitose kitasizwe irangi, iyo RNA ntigaragara. Irangi rya supravital ryinjira mu karemangingo kakiri kazima, rigatuma iyo RNA yirema cyangwa yegerana ikaba utubumbe n'ududodo tw'ubururu, ari byo bikoresho bisa n'urusobe rw'ududodo bigaragara iyo hakoreshejwe mikorosikopi ifite objektifu yibizwa mu mavuta. Bityo, urwo rusobe rubaho kubera gusiga irangi; si urwego rusanzwe rwubatse rutegereje gusa guhabwa ibara. Ni yo mpamvu ibintu bihinduka mu mabwiriza y'uburyo bukoreshwa, nk'ubukana bw'irangi, umuti ugumisha pH, igipimo cy'amaraso ugereranyije n'irangi, ubushyuhe n'igihe, bifite akamaro kangana n'izina ry'irangi.

Icyo bivuze mu bikorwa ni uko intambwe zigomba gukurikizwa uko ziteye: banza uvange amaraso yashyizwemo umuti ubuza kuvura n'irangi, ureke bimare igihe mu bihe byagenwe, hanyuma ubisige ku birahure ubisuzume bitafatishejwe kandi bitasizwe irangi ryunganira. Gufatisha mbere yo gusiga, cyangwa gukoresha irangi ryunganira rihisha ududodo dufite ibara ridakomeye, byangiza amakuru kubara bishingiraho.

Amabwiriza yo gusiga retikulocyte hakoreshejwe new methylene blue

Nta mvange imwe ikoreshwa hose, bityo vuga iyakoreshejwe. Uburyo butatu bwanditswe bugaragaza uko imvange zishobora gutandukana.

Uburyo busanzwe bwo mu bwoko bwa Brecher bukoresha new methylene blue iri ku gipimo cya hafi 0.5 percent hamwe na potassium oxalate, bikavangwa n'ingano ingana y'amaraso arimo EDTA yavanzwe neza, hanyuma bikamara hafi 10 kugeza kuri 15 minutes ku bushyuhe bwo mu cyumba. Nyuma yo kongera kubivanga buhoro, hasigwa amaraso ku birahure mu buryo butanga agace kananutse kagenda kagabanuka, akumishwa n'umwuka, hanyuma agasuzumwa atafatishejwe hakoreshejwe objektifu ya 100x yibizwa mu mavuta. Amabwiriza ya NCBI yo kubara retikulocyte asobanura izi ntambwe n'urubibi rw'agace ko kubariramo rukoreshwa muri ubu buryo.

Uburyo bwa Dacie na Lewis bukoresha new methylene blue iri ku gipimo cya 1.0 percent mu muti w'umunyu urimo citrate, ikavangwa n'ingano y'amaraso iyikubye 2 kugeza kuri 4, hanyuma bikagumishwa kuri 37 C mu gihe cya 15 kugeza kuri 20 minutes mbere yo kubisiga ku birahure. Mu ishyirwa mu bikorwa riheruka ryo guhuza uburyo bwa CLSI H44, hakoreshejwe umuti ufite irangi rito kurushaho, ku gipimo cya 0.1 percent, mu muti w'umunyu ufite phosphate igumisha pH kuri 7.4, ukamara kuri 20 C mu gihe cya 10 minutes, hatabayeho gufatisha, nk'uko byatangajwe mu bushakashatsi bwa 2026 bwo guhuza uburyo bwa NMB.

Amategeko yo kubara ni amwe muri ubu buryo bwose. Bara akaremangingo nka retikulocyte iyo karimo nibura utubumbe tubiri twihariye twasizwe ubururu cyangwa uduce tubiri tw'ududodo, bityo ukirengagiza irangi ryifashe ahantu hamwe gusa. Mu kazi gasanzwe, bara nibura uturemangingo dutukura 1,000, maze utangaze retikulocyte nk'ijanisha. Iyo ijanisha riri hasi, imibare isaba kubara uturemangingo twinshi kurushaho. Isuzuma ry'uburyo bwa CLSI H44 rivuga ko iyo retikulocyte ziri hafi ya 1 percent, hakenerwa uturemangingo tugera kuri 4,400 kugira ngo ihindagurika ryo kubara rigabanuke rigere kuri 5 percent, nk'uko biri mu isuzuma rya CLSI H44 kuri De Gruyter. Disiki ya Miller ishyirwa mu ndorerwamo y'ijisho ya mikorosikopi yorohereza kubara uturemangingo twinshi. Yungurura irangi cyangwa utegure irishya, kuko irangi ryiremyemo utubumbe rishobora kwitiranywa n'utubumbe two mu turemangingo.

Utubumbe twa Heinz muri icyo cyitegererezo kimwe

Icyitegererezo kimwe cyasizwe mu buryo bwa supravital gishobora no kugaragaza utubumbe twa Heinz, ari two twegeranya rya hemoglobin yagize okisidasiyo kandi imiterere yayo igahinduka, aho kuba ibisigazwa bya RNA. Mu byitegererezo bya new methylene blue, tugaragara nk'utubumbe tuzengurutse, akenshi ari kamwe kandi kari hafi y'agahu k'akaremangingo, dufite ubururu bworoshye kurusha ibikoresho bisa n'urusobe rw'ududodo. Ntituguma kugaragara neza nyuma yo gufatisha no gusiga irangi ryunganira, bityo suzuma icyasizwe ku kirahure kitafatishejwe. Amabwiriza aheruka yakoresheje 20 microliters z'amaraso arimo EDTA na 20 microliters za new methylene blue iri ku gipimo cya 1 percent, bikamara 15 minutes mu bihe byagenwe, maze habarwa uturemangingo turimo utubumbe twa Heinz muri erythrocytes 250, nk'uko biri mu bushakashatsi bwo kuri PMC bwatangaje ubwo buryo.

Itandukaniro ry'ibintu bibiri ririnda gusoma nabi ibigaragara. Icya mbere, methyl violet na crystal violet bisiga utubumbe twa Heinz ibara rikomeye kurusha new methylene blue, nk'uko byavuzwe mu nyandiko ya ScienceDirect yerekeye utubumbe twa Heinz. New methylene blue ni yo yorohereza akazi igihe retikulocyte n'utubumbe twa Heinz bisuzumirwa hamwe, ariko si yo irangi rikomeye kurusha ayandi yagenewe by'umwihariko utubumbe twa Heinz. Icya kabiri, utubumbe twa HbH two mu turemangingo ku bantu bafite ikiranga cya alpha thalassemia dutanga utudomo duto twinshi mu karemangingo kose nyuma yo gusiga mu buryo bwa supravital, mu gihe utubumbe twa Heinz dusanzwe tuba duke, tunini kurushaho kandi akenshi dufatanye n'agahu k'akaremangingo. Irangi ryifashe ku kirahure ni ikintu cya gatatu gishobora kwitiranywa na two: riba ritari ku rwego rumwe n'uturemangingo aho mikorosikopi irebera neza, kandi rihinduka hagati y'uduce turebwamo.

Brilliant cresyl blue ugereranyije na new methylene blue

Aya marangi yombi akoreshwa mu gusiga retikulocyte zikiri nzima, ariko ibipimo ngenderwaho bishyira imbere rimwe muri yo. Uburyo bwa CLSI H44-A2 bwo gusuzuma retikulocyte, bwateguwe ku bufatanye n'Inama Mpuzamahanga Ishinzwe Ibipimo Ngenderwaho mu Bumenyi bw'Amaraso kandi buri ku rupapuro rw'igipimo ngenderwaho cya CLSI H44, bugena ko gusiga amaraso arimo EDTA na new methylene blue ari bwo buryo bw'intoki bwo kugereranyaho. Inyigisho zisanzwe z'ubumenyi bw'amaraso zihuriza ku miterere y'ibigaragara: new methylene blue isiga ududodo ibara rikomeye kandi risa neza hose, mu gihe brilliant cresyl blue ihindagurika kurushaho bitewe n'icyiciro cy'irangi ryakozwe n'icyitegererezo.

Icyo cyemezo kigomba guherekezwa n'icyitonderwa. Amabwiriza ya ICSH yo gusuzuma ibikoresho bipima amaraso yemera gukoresha mikorosikopi mu buryo bw'intoki hamwe na new methylene blue nk'uburyo fatizo bwo kugereranyaho, ariko avuga ko bushingira ku buryo umuntu abona kandi ko budatanga imibare ihamye cyane; uburyo fatizo bwa flow cytometry ni bwo bushyirwa imbere aho buboneka. Igereranya ryakozwe mu 2022 ku byitegererezo 100 ryasanze brilliant cresyl blue ifite ukubogama kuke kurusha new methylene blue iyo hagereranyijwe n'igikoresho cyapimaga amaraso muri ubwo bushakashatsi, nk'uko byatangajwe mu igereranya ry'uburyo bwo gusiga riri kuri PMC. Incamake ifite ishingiro ku banyeshuri ni iyi: koresha new methylene blue nk'uburyo bw'intoki bugengwa n'ibipimo ngenderwaho bwo kugereranyaho, genzura uburyo ubwo ari bwo bwose bwa brilliant cresyl blue bukoreshwa aho ukorera ubigereranya na yo cyangwa n'ibikoresho byikoresha, kandi uteganye ihindagurika ryagutse mu bisubizo bya mikorosikopi ikoreshwa n'intoki, uko uburyo bwaba bumeze kose. Inyandiko za kera ku gushyiraho ibipimo ngenderwaho na zo zashyigikiraga Azure B yatunganyijwe nk'ubundi buryo bufite ubuziranenge bwa shimi kurushaho, kuko ibyiciro by'amarangi yagurishwaga byatandukanyaga, nk'uko biri mu bushakashatsi bwa Marshall kuri Azure B. Ibyo bisobanura impamvu urwego rw'ubuziranenge bw'inyongeramusaruro rukigaruka mu biganiro ku buryo bukoreshwa.

Kwirinda amakosa asanzwe

Amakosa ane ni yo atera ibyinshi mu byitegererezo bidatanga ibyifuzwa. Gutumiza methylene blue isanzwe aho gutumiza new methylene blue N bitera ibara ridakomeye bitewe n'imiterere ya shimi, si uburyo yakoreshejwe. Gufatisha cyangwa gusiga irangi ryunganira mbere yo kureba bikuraho ibimenyetso bitangwa n'uburyo bwa supravital. Kubara utubumbe tw'irangi ryiremyemo nk'utwo mu turemangingo byongera igisubizo bidakwiye; itegeko ry'utubumbe tubiri no gukoresha irangi ryayunguruwe birabikumira. Kubara uturemangingo duke cyane igihe ijanisha rya retikulocyte riri hasi bitanga umubare usa n'uwizewe ariko ufite intera y'icyizere yagutse. Buri kosa rifite ikimenyetso kimwe: ukoresha mikorosikopi ayishyiraho ikosa kandi ibyabaye byaragenwe n'intambwe yo gukoresha inyongeramusaruro.