Kuongorora maleukocyte muropa rose ongororo yenguva dzose mukiriniki lab kana bhangi reropa.Iko kusungirirwa uye kushanda kweiyo leukocytes ndiyo yakakosha index sekutonga kwehutano hwekuchengetedza ropa.Kunze kweleukocyte, ropa rose rine nhamba yakawanda yeplatelet, masero matsvuku eropa, kana masero emarara, izvo zvinoita kuti zvisakwanise kuongorora ropa rose zvakananga pasi pe microscope kana kuti chiedza chemunda wemasero.Nzira dzakajairika dzekuverenga masero machena eropa dzinosanganisira RBC lysis process, inopedza nguva.
AOPI Dual-fluoresces kuverenga imhando yeassay inoshandiswa kuona kusungwa kwesero uye kushanda.Mhinduro isanganiswa yeacridine orange (iyo green-fluorescent nucleic acid stain) uye propidium iodide (iyo red-fluorescent nucleic acid stain).Propidium iodide (PI) idhayi yekusarudzika membrane inongopinda mumasero ane membrane yakakanganiswa, nepo acridine orenji inokwanisa kupinda mumasero ese muhuwandu.Kana madhayi ese ari maviri ari munucleus, propidium iodide inokonzera kuderedzwa kweacridine orange fluorescence nefluorescence resonance energy transfer (FRET).Nekuda kweizvozvo, masero ane nucleated ane intact membranes anoshatisa fluorescent green uye anoverengerwa semhenyu, nepo nucleated maseru ane membrane akakanganiswa anongosvibisa fluorescent tsvuku uye anoverengwa seakafa kana uchishandisa Countstar® Rigel system.
Countstar Rigel mhinduro yakanakira kune akawanda akaomarara ehuwandu hwehuwandu hwemaitiro assays, inogonesa kukurumidza kuongorora machena-eropa-masero muropa rose.
Maitiro ekuedza:
1.Tora 20 µl yesampuli yeropa uye dilute sampuro mu180 µl yePBS.
2.Wedzera 12µl AO/PI mhinduro mu12µl sampuli, zvinyoronyoro yakasanganiswa nepipette;
3.Dhirora 20µl musanganiswa mukamuri siraidhi;
4.Bvumira masero kuti agare mukamuri kweinenge 1 miniti;
5.Insect the slide kupinda Countstar FL chiridzwa;
6.Sarudza "AO / PI Viability" assay, ipapo Isa Sample ID yemuenzaniso uyu.
7.Sarudza Dilution ratio, Cell Type, iyo tinya 'Mhanya' kuti utange bvunzo.
Yambiro: AO uye PI inogona kukonzera kenza.Zvinokurudzirwa kuti anoshanda apfeke midziyo yekudzivirira (PPE) kudzivirira zvakanangana neganda nemeso.
Mhedzisiro:
1. Bright Field Image yeropa rose
Mumufananidzo wakajeka weropa rose, maWBC haaonekwe pakati pesero dzvuku reropa.(Mufananidzo 1)
Mufananidzo 1 Mufananidzo wemunda wakajeka weropa rose.
2. Fluorescence Mufananidzo weropa rose
Iyo AO nePI dhayi zvese zvinosvibisa DNA musero nucleus yemasero.Naizvozvo, maPlatelets, masero matsvuku eropa, kana marara emasero haakwanise kukanganisa kutariswa kweleukocytes uye mhedzisiro.Rarama leukocytes (Green) uye akafa leukocyte (Red) anoonekwa zviri nyore mumifananidzo yefluorescence.(Mufananidzo 2)
Mufananidzo 2 Fluorescence Mifananidzo yeropa rose.(A).Mufananidzo weAO Channel;(B) Mufananidzo wePI Channel;(C) Batanidza mifananidzo yeAO nePI Channel.
3. Kuisa pfungwa uye kubudirira kwe leukocytes
Iyo Countstar FL software inoverenga otomatiki maseru ezvikamu zvitatu zvekamuri uye inoverengera kukosha kweavhareji yehuwandu hweWBC cell count (1202), concentration (1.83 x 106 masero/ml), uye % kugona (82.04%).Iyo yese mifananidzo yeropa uye data inogona kutumirwa kunze nyore sePDF, Mufananidzo kana Excel kuti uwedzere kuongororwa kana kuchengetedza data.
Mufananidzo 3 Screenshot ye Countstar Rigel Software
