ʻO ka nānā ʻana i nā leukocytes i loko o ke koko holoʻokoʻa he hoʻomaʻamaʻa maʻamau i ke keʻena lapaʻau a i ʻole ka waihona koko.ʻO ka noʻonoʻo ʻana a me ka hiki ʻana o nā leukocytes ka mea nui e like me ka mālama ʻana i ke koko.Ma waho aʻe o ka leukocyte, loaʻa i ke koko holoʻokoʻa ka nui o nā platelets, nā ʻulaʻula ʻulaʻula, a i ʻole nā ʻōpala cellular, kahi i hiki ʻole ai ke ʻimi pono i ke koko holoʻokoʻa ma lalo o ka microscope a i ʻole nā counter cell field bright.ʻO nā ʻano hana maʻamau e helu i nā keʻokeʻo keʻokeʻo e pili ana i ke kaʻina RBC lysis, ʻo ia ka manawa e pau ai.
ʻO ka helu AOPI Dual-fluoresces ka ʻano assay i hoʻohana ʻia no ka ʻike ʻana i ka neʻe ʻana o ke kelepona a me ka hiki.ʻO ka hoʻonā ʻana he hui pū ʻana o ka ʻalani acridine (ka ʻakika nucleic acid green-fluorescent) a me ka propidium iodide (ka ʻulaʻula nucleic acid stain).ʻO ka propidium iodide (PI) he mea hoʻoheheʻe hoʻokaʻawale membrane e komo wale i nā pūnaewele me nā membrane i hoʻopaʻa ʻia, ʻoiai hiki i ka ʻalani acridine ke komo i nā cell āpau i kahi heluna.Ke loaʻa nā wai ʻelua i loko o ka nucleus, ʻo propidium iodide ka mea e hoʻemi ai i ka fluorescence ʻalani acridine ma o ka hoʻololi ʻana i ka ikehu resonance (FRET).ʻO ka hopena, ʻo nā pūnaewele nucleated me nā membrane paʻa i hoʻopaʻa i ka ʻōmaʻomaʻo ʻōmaʻomaʻo a helu ʻia he ola, ʻoiai nā cell nucleated me nā membrane i hoʻopaʻapaʻa wale ʻia ka ʻulaʻula fluorescent a helu ʻia he make ke hoʻohana i ka ʻōnaehana Countstar® Rigel.
He hopena maikaʻi ʻo Countstar Rigel no ka nui o ka heluna helu helu kanaka paʻakikī, hiki ke hoʻopaʻa koke i nā keʻokeʻo-koko-keʻokeʻo i loko o ke koko holoʻokoʻa.
Kaʻina hana hoʻokolohua:
1. E lawe i 20 µl o ka laʻana koko a hoʻoheheʻe i ka hāpana i 180 µl o PBS.
2. Hoʻohui i ka 12µl AO / PI solution i loko o ka hāpana 12µl, hui mālie me ka pipette;
3. E huki i ka hui ʻana o 20µl i loko o ke keʻena slide;
4. E hoʻokuʻu i nā pūnaewele e noho i loko o ke keʻena no kahi o 1 minuke;
5. E ʻoki i ka paheʻe i loko o ka mea kani Countstar FL;
6. E koho i ka "AO / PI Viability" assay, a laila e hoʻokomo i ka Sample ID no kēia laʻana.
7.Select Dilution ratio, Cell Type, ke kaomi 'Run' e hoʻomaka i ka hoʻāʻo.
E akahele: ʻO AO a me PI he mea make kino.Manaʻo ʻia e hoʻohana ka mea hoʻohana i nā lako pale pilikino (PPE) e pale aku i ka pili pono ʻana me ka ʻili a me nā maka.
Ka hopena:
1. Bright Field Image o ke koko holookoa
I ke kiʻi ʻālohilohi o ke koko holoʻokoʻa, ʻaʻole ʻike ʻia nā WBC ma waena o ke koko ʻulaʻula.(Kiʻi 1)
Kiʻi 1 Kiʻi kahua mālamalama o ke koko holoʻokoʻa.
2. Fluorescence Kiʻi o ke koko holoʻokoʻa
ʻO ka dye AO a me ka PI he mau kiko DNA ʻelua i loko o ka nucleus cell of cell.No laila, ʻaʻole hiki i nā Platelets, nā keʻena koko ʻulaʻula, a i ʻole nā ʻōpala cellular ke hoʻopilikia i ka ʻike leukocytes a me ka hopena ola.ʻIke maʻalahi nā leukocytes ola (Green) a me nā leukocytes make (Red) i nā kiʻi fluorescence.(Kiʻi 2)
Kiʻi 2 Fluorescence Nā kiʻi o ke koko holoʻokoʻa.(A).Kii o AO Channel;(B) Kiʻi o PI Channel;(C) Hoʻohui i nā kiʻi o AO a me PI Channel.
3. Ka nui a me ka ola o nā leukocytes
Helu 'akomi ka lako polokalamu Countstar FL i nā keena o nā ʻāpana keʻena ʻekolu a helu i ka waiwai awelika o ka helu ʻana o ka cell WBC (1202), ka manaʻo (1.83 x 106 cell/ml), a me ka % viability (82.04%).Hiki ke lawe maʻalahi ʻia nā kiʻi koko holoʻokoʻa a me nā ʻikepili ma ke ʻano he PDF, Image a Excel paha no ka loiloi hou a i ʻole ka waihona ʻikepili.
Kiʻi 3 Kiʻi kiʻi o ka polokalamu Countstar Rigel
