Sistemụ Countstar na-ejikọta cytometer onyonyo na counter cell n'ime otu ngwa bench-top.Ngwa a na-akwalite, kọmpat, na sistemụ onyonyo akpaaka na-enye ihe ngwọta niile maka nyocha mkpụrụ ndụ kansa, gụnyere ọnụ ọgụgụ cell, viability (AO/PI, trypan blue), apoptosis (Annexin V-FITC/PI), cell. okirikiri (PI), na nfefe GFP/RFP.
Abstract
Ọrịa cancer bụ otu n'ime isi ihe na-akpata ọnwụ n'ụwa niile, na mmepe nke usoro ọgwụgwọ ọrịa cancer ọhụrụ dị oke mkpa.Cell cancer bụ isi ihe nyocha nke ọrịa kansa, ọ dị mkpa ka enyocha ozi dị iche iche site na mkpụrụ ndụ kansa.Mpaghara nyocha a chọrọ nyocha sel ngwa ngwa, ntụkwasị obi, dị mfe na nke zuru oke.Sistemụ Countstar na-enye usoro ngwọta dị mfe maka nyocha mkpụrụ ndụ kansa.
Mụọ Apoptosis Cell Cancer nke Countstar Rigel
A na-ejikarị nyocha apoptosis eme ihe n'ọtụtụ ụlọ nyocha maka ebumnuche dị iche iche site na nyocha ahụike nke omenala cell ruo n'ịtụle nsi nke otu panel nke ogige.
Apoptosis assay bụ ụdị eji achọpụta pasentị apoptosis nke mkpụrụ ndụ site na usoro ntụpọ Annexin V-FITC/PI.Annexin V na-ejikọta na phosphatidylserine (PS) na cell apoptosis n'oge ma ọ bụ cell necrosis.PI na-abanye naanị necrotic / sel apoptotic nke oge ikpeazụ.(Foto nke 1)
A: apoptosis nke mbụ Annexin V (+), PI (-)
B: apoptosis mbubreyo Annexin V (+), PI (+)
Ọnụọgụgụ 1: Nkọwa gbasara foto Countstar Rigel (5 x na-ebuwanye ibu) nke mkpụrụ ndụ 293, ejiri Annexin V FITC na PI gwọọ ya.
Nyochaa okirikiri cell nke mkpụrụ ndụ kansa
Selụ okirikiri ma ọ bụ okirikiri nkewa cell bụ usoro ihe omume na-eme na cell na-eduga na nkewa ya na ntugharị DNA ya (DNA replication) iji mepụta mkpụrụ ndụ ụmụ nwanyị abụọ.N'ime mkpụrụ ndụ nwere oghere, dị ka ọ dị na eukaryotes, a na-ekewakwa okirikiri cell n'ime oge atọ: interphase, usoro mitotic (M), na cytokinesis.Propidium iodide (PI) bụ ihe na-agba agba nuklia nke a na-ejikarị tụọ okirikiri cell.N'ihi na agba ahụ enweghị ike ịbanye na mkpụrụ ndụ dị ndụ, a na-edozi mkpụrụ ndụ ahụ na ethanol tupu agbapụta ya.Selụ niile na-emezi ntụpọ.Selụ na-akwado maka nkewa ga-enwe ọnụọgụ DNA na-abawanye ma gosipụta nha nha nha nha.A na-eji ndịiche dị na ike fluorescence iji chọpụta pasentị nke mkpụrụ ndụ na akụkụ nke ọ bụla nke okirikiri cell.Countstar nwere ike weghara onyonyo a, a ga-egosipụta nsonaazụ ya na ngwa ngwa FCS.(Foto nke 2)
Ọgụgụ 2: MCF-7 (A) na 293T (B) ka ejiri ngwa nchọpụta okirikiri cell nwere PI, Countstar Rigel kpebiri nsonaazụ ya, wee nyochaa ya site na FCS express.
Mkpebi nfefe GFP na cell
N'oge bioprocess, a na-ejikarị GFP ejikọta ya na protein recombinant dị ka ihe ngosi.Kpebisie ike na GFP fluorescent nwere ike gosipụta nkwupụta protein ebumnuche.Countstar Rigel na-enye nyocha ngwa ngwa ma dị mfe maka ịnwale mbufe GFP yana ike.Ejiri Propidium iodide (PI) na Hoechst 33342 nwere ntụpọ mkpụrụ ndụ iji kọwaa ọnụ ọgụgụ sel nwụrụ anwụ na mkpokọta ọnụ ọgụgụ cell.Countstar Rigel na-enye usoro ọsọ ọsọ, ọnụọgụ maka ịlele arụmọrụ okwu GFP na ịdị adị n'otu oge.(Foto nke 4)
Ọgụgụ 4: Selụ dị site na iji Hoechst 33342 (acha anụnụ anụnụ) na pasent nke GFP na-egosipụta mkpụrụ ndụ (green) nwere ike ikpebi ngwa ngwa.A na-eji propidium iodide (PI; acha ọbara ọbara).
Viability na ọnụ ọgụgụ cell
AO/PI Dual-fluoresces na-agụta bụ ụdị nyocha ejiri maka ịchọpụta mkpokọta cell, ịdị adị.Ọ kewara n'ime ọnụ ọgụgụ ahịrị cell na ọnụọgụ mkpụrụ ndụ nke mbụ dịka ụdị sel dị iche iche si dị.Ihe ngwọta nwere nchikota nke green-fluorescent nucleic acid unyi, acridine oroma, na redfluorescent nucleic acid unyi, propidium iodide.Propidium iodide bụ ụcha mwepu nke akpụkpọ ahụ nke na-abanye naanị n'ime mkpụrụ ndụ nwere membranes mebiri emebi ebe oroma acridine na-abanye n'ime sel niile dị na ọnụ ọgụgụ mmadụ.Mgbe agbaji abụọ dị na nucleus, propidium iodide na-eme ka mbelata nke acridine oroma fluorescence site na fluorescence resonance energy transfer (FRET).N'ihi nke a, mkpụrụ ndụ nucleated nwere akpụkpọ ahụ na-adịghị emebi emebi na-acha akwụkwọ ndụ akwụkwọ ndụ na-acha akwụkwọ ndụ akwụkwọ ndụ, ebe a na-agụkwa ya dị ka ndị dị ndụ, ebe sel ndị nwere nsogbu mebie na-acha uhie uhie na-acha uhie uhie na-agụkwa dị ka ndị nwụrụ anwụ mgbe ha na-eji usoro Countstar Rigel.Ihe anaghị emebi emebi dị ka mkpụrụ ndụ ọbara uhie, platelet na irighiri ihe anaghị apụta, ngwa Countstar Rigel na-eleghara ya anya.(Foto nke 5)
Onyonyo 5: Countstar emeziwanyela usoro ntụpọ abụọ-fluorescence maka mfe, mkpebi ziri ezi nke itinye uche na ike PBMC.Enwere ike iji Countstar Rigel nyochaa ihe nlele ndị nwere AO/PI
