Name: Nekroz, Apoptoz ve Trombosit Aktivasyonu Özellikleri Sergileyen Prokoagülan Trombositlerin Kapsamlı Analizi (Video) | JoVE Journal
Uploaded: 2025-05-23T14:00:00.000+00:00
Duration: 4 min 37 s
Description: Öğrenin, prokoagülan trombositleri nekroz, apoptoz ve aktivasyon özellikleriyle analiz etme yöntemini. Bu protokol, tromboz riski değerlendirmesi ve tedavi hedefleri için metabolik yolakları incelemeye yardımcı olur.

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<li>Kholmukhamedov, A., Janecke, R., Choo, H. -J., Jobe, S. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+mitochondrial+calcium+uniporter+regulates+procoagulant+platelet+formation%5BTitle%5D)+AND+%22J+Thromb+Haemost%22%5BJournal%5D)">The mitochondrial calcium uniporter regulates procoagulant platelet formation.</a> J Thromb Haemost. 16 (11), 2315-2321 (2018).</li>
<li>Choo, H. -J., Saafir, T. B., Mkumba, L., Wagner, M. B., Jobe, S. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Mitochondrial+calcium+and+reactive+oxygen+species+regulate+agonist-initiated+platelet+phosphatidylserine+exposure%5BTitle%5D)+AND+%22Arterioscler+Thromb+Vasc+Biol%22%5BJournal%5D)">Mitochondrial calcium and reactive oxygen species regulate agonist-initiated platelet phosphatidylserine exposure.</a> Arterioscler Thromb Vasc Biol. 32 (12), 2946-2955 (2012).</li>
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<li>Schoenwaelder, S. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Two+distinct+pathways+regulate+platelet+phosphatidylserine+exposure+and+procoagulant+function%5BTitle%5D)+AND+%22Blood%22%5BJournal%5D)">Two distinct pathways regulate platelet phosphatidylserine exposure and procoagulant function.</a> Blood. 114 (3), 663-666 (2009).</li>
<li>Abbasian, N., Millington-Burgess, S. L., Chabra, S., Malcor, J. -D., Harper, M. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Supramaximal+calcium+signaling+triggers+procoagulant+platelet+formation%5BTitle%5D)+AND+%22Blood+Adv%22%5BJournal%5D)">Supramaximal calcium signaling triggers procoagulant platelet formation.</a> Blood Adv. 4 (1), 154-164 (2020).</li>
<li>Le, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((An+inner+activation+gate+controls+TMEM16F+phospholipid+scrambling%5BTitle%5D)+AND+%22Nat+Commun%22%5BJournal%5D)">An inner activation gate controls TMEM16F phospholipid scrambling.</a> Nat Commun. 10 (1), 1846(2019).</li>
<li>Yang, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((TMEM16F+forms+a+Ca2%2B-activated+cation+channel+required+for+lipid+scrambling+in+platelets+during+blood+coagulation%5BTitle%5D)+AND+%22Cell%22%5BJournal%5D)">TMEM16F forms a Ca2+-activated cation channel required for lipid scrambling in platelets during blood coagulation.</a> Cell. 151 (1), 111-122 (2012).</li>
<li>Mattheij, N. J. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Dual+mechanism+of+integrin+closure+in+procoagulant+platelets%5BTitle%5D)+AND+%22J+Biol+Chem%22%5BJournal%5D)">Dual mechanism of integrin closure in procoagulant platelets.</a> J Biol Chem. 288 (19), 13325-13336 (2013).</li>
<li>Liu, F., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Mitochondrially+mediated+integrin+%CE%B1iIb%CE%B23+protein+inactivation+limits+thrombus+growth%5BTitle%5D)+AND+%22J+Biol+Chem%22%5BJournal%5D)">Mitochondrially mediated integrin αiIbβ3 protein inactivation limits thrombus growth.</a> J Biol Chem. 288 (42), 30672-30681 (2013).</li>
<li>van Kruchten, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Both+TMEM16F-dependent+and+TMEM16F-independent+pathways+contribute+to+phosphatidylserine+exposure+in+platelet+apoptosis+and+platelet+activation%5BTitle%5D)+AND+%22Blood%22%5BJournal%5D)">Both TMEM16F-dependent and TMEM16F-independent pathways contribute to phosphatidylserine exposure in platelet apoptosis and platelet activation.</a> Blood. 121 (10), 1850-1857 (2013).</li>
<li>Goelz, N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Platelets+express+adaptor+proteins+of+the+extrinsic+apoptosis+pathway+and+can+activate+caspase-8%5BTitle%5D)+AND+%22PLOS+ONE%22%5BJournal%5D)">Platelets express adaptor proteins of the extrinsic apoptosis pathway and can activate caspase-8.</a> PLOS ONE. 16 (1), e0244848(2021).</li>
<li>Shlomovitz, I., Speir, M., Gerlic, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Flipping+the+dogma+-+phosphatidylserine+in+non-apoptotic+cell+death%5BTitle%5D)+AND+%22Cell+Commun+Signal%22%5BJournal%5D)">Flipping the dogma - phosphatidylserine in non-apoptotic cell death.</a> Cell Commun Signal. 17 (1), 139(2019).</li>
<li>Nadine, J. A. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Coated+platelets+function+in+platelet-dependent+fibrin+formation+via+integrin+%CE%B1IIb%CE%B23+and+transglutaminase+factor+XIII%5BTitle%5D)+AND+%22Haematologica%22%5BJournal%5D)">Coated platelets function in platelet-dependent fibrin formation via integrin αIIbβ3 and transglutaminase factor XIII.</a> Haematologica. 101 (4), 427-436 (2016).</li>
<li>Michelson, A. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Platelet+activation+by+thrombin+can+be+directly+measured+in+whole+blood+through+the+use+of+the+peptide+GPRP+and+flow+cytometry%3A+Methods+and+clinical+applications%5BTitle%5D)+AND+%22Blood+Coagul+Fibrinolysis%22%5BJournal%5D)">Platelet activation by thrombin can be directly measured in whole blood through the use of the peptide GPRP and flow cytometry: Methods and clinical applications.</a> Blood Coagul Fibrinolysis. 5 (1), 121-131 (1994).</li>
<li>Taylor, M. R., Couto, J. R., Scallan, C. D., Ceriani, R. L., Peterson, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Lactadherin+%28formerly+BA46%29%2C+a+membrane-associated+glycoprotein+expressed+in+human+milk+and+breast+carcinomas%2C+promotes+Arg-Gly-Asp+%28RGD%29-dependent+cell+adhesion%5BTitle%5D)+AND+%22DNA+Cell+Biol%22%5BJournal%5D)">Lactadherin (formerly BA46), a membrane-associated glycoprotein expressed in human milk and breast carcinomas, promotes Arg-Gly-Asp (RGD)-dependent cell adhesion.</a> DNA Cell Biol. 16 (7), 861-869 (1997).</li>
<li>Andersen, M. H., Graversen, H., Fedosov, S. N., Petersen, T. E., Rasmussen, J. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Functional+analyses+of+two+cellular+binding+domains+of+bovine+lactadherin%5BTitle%5D)+AND+%22Biochemistry%22%5BJournal%5D)">Functional analyses of two cellular binding domains of bovine lactadherin.</a> Biochemistry. 39 (20), 6200-6206 (2000).</li>
<li>Tan, C. W., Bourcy, M., Pasalic, L., Chen, V. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Flow+cytometry+assessment+of+procoagulant+platelets+using+a+dithiol-reactive+probe+functional+disulfide+bonds%3A+Methods+and+protocols%5BTitle%5D)+AND+%22Methods+Mol+Biol%22%5BJournal%5D)">Flow cytometry assessment of procoagulant platelets using a dithiol-reactive probe functional disulfide bonds: Methods and protocols.</a> Methods Mol Biol. 1967, 305-321 (2019).</li>
</ol>

<table><tbody><tr><td>Asit Sitrat Dekstroz (ACD) çözeltisi (1000 mL için)</td><td></td><td></td> <td>Tri- Sodyum Sitrat- &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 22 g Sitrik Asit-&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 8 g Dekstroz-&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 24,5 g Su-&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 1000 mL'ye kadar hacim oluşturun&nbsp;</td></tr><tr><td>Alexa Fluor 488 protein etiketleme kiti</td><td>Invitrogen</td><td>A10235</td><td></td></tr><tr><td>APC Fare Anti-İnsan CD62P</td><td>BD Pharmingen</td><td>550888</td><td></td></tr><tr><td>Sığır Protrombin</td><td>Prolytix</td><td>BCP-1010</td><td></td></tr><tr><td>Tampon A (Trombosit Hazırlama)</td><td></td><td></td><td>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; M.W&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; Conc. 1X&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 100 mL 10X çözeltisi için HEPES&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 238.30&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 20 mM&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 4.766 g NaCl&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 58.44&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 134 mM&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 7.83 g KCl&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 74,55&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 2.9 mM&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 216.19 mg MgCl2&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 203.30&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 1 mM&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 203.30 mg NaH2PO4&nbsp; &nbsp; &nbsp; 156.01&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 0.34 mM&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 53.04 mg NaHCO3&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 84 01&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 12 mM & nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 1.01 g pH'ı 6.2 'ye ayarladıktan sonra 100 mL'ye kadar su Aşağıdakilerle desteklenmesi gereken 1X solüsyonu elde etmek için trombosit hazırlamadan hemen önce 10X solüsyonu 1:10 (h/h) seyreltin &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; Conc. 1X&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 1 ml 1X çözelti için EGTA &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 1 mM &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 10 &mu ekleyin; L (1:100 v/d) 100 mM EGTA&nbsp; Glikoz &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 5 mM & nbsp; &nbsp; &nbsp; &nbsp; 5 &mu ekleyin; L (1:200 v/h) 1 M glikoz PGE1 & nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 3 &mu; M & nbsp; &nbsp; &nbsp; &nbsp; 3 &mu ekleyin; L (1:333 v/h) 1 mM PGE1 çözeltisi</td></tr><tr><td>Tampon B (Trombosit Hazırlama)</td><td></td><td></td><td>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; M.W&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; Conc. 1X&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 100 mL 10X çözeltisi için HEPES&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 238.30&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 20 mM&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 4.766 g NaCl&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 58.44&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 134 mM&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 7.83 g KCl&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 74,55&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 2.9 mM&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 216.19 mg MgCl2&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 203.30&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 1 mM&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 203.30 mg NaH2PO4&nbsp; &nbsp; &nbsp; 156.01&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 0.34 mM&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 53.04 mg NaHCO3&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 84 01&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 12 mM & nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 1.01 g pH'ı 7.4 'ye ayarladıktan sonra 100 mL'ye kadar su Aşağıdakilerle desteklenmesi gereken 1X solüsyonu elde etmek için trombosit hazırlamadan hemen önce 10X solüsyonu 1:10 (h/h) seyreltin &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; Conc. 1X&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 1 ml 1X çözelti için Glikoz &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; 5 mM & nbsp; &nbsp; &nbsp; &nbsp; 5 &mu ekleyin; L (1:200 v/h) 1 M glikoz&nbsp;</td></tr><tr><td>CaCl2 (0.5 M) (50ml için)</td><td></td><td></td><td>CaCl2.2H2O'nun moleküler ağırlığı = 147.02 3.675 g CaCl2.2H2O'yu 50 ml Milli Q su içinde çözün</td></tr><tr><td>CellEvent Caspase-3/7 Tespit Reaktifleri Yeşil</td><td>Invitrogen</td><td>C10423</td><td>Apoptoz tespit reaktifi</td></tr><tr><td>Convulxin</td><td>Enzo Yaşam Bilimleri</td><td>ALX-350-100</td><td></td></tr><tr><td>EDTA (0.5 M; pH 8) (100 mL için)</td><td></td><td></td><td>EDTA Na2.2H2 O'nun Moleküler Ağırlığı: 372.24g 18.612 g ağırlığında ve 50 ml Milli Q suda süspanse edin ve pH'ı kontrol edin (pH ~ 4) Karıştırarak yavaşça 10N NaOH ekleyin ve pH'ı izleyin. EDTA, pH 7 civarında çözünmeye başlar ve pH 8. 'da tamamen çözünür. Milli Q su ile hacmi 100 ml'ye kadar tamamlayın.</td></tr><tr><td>EGTA (100 mM; pH 7.4) (100 mL için)</td><td></td><td></td><td>Molekül Ağırlığı: 380.4 50 ml Milli Q suya 3.804 g EGTA ekleyin ve pH'ı (pH~3) kontrol edin. Karıştırırken damla damla 10 N NaOH ekleyin ve pH 'ı izleyin EGTA pH 7.0'da çözünür hale gelir (yaklaşık) pH'ı 7.4 'a ayarlayın Milli Q su</td></tr> <tr><td>ile hacmi 100 ml'ye kadar tamamlayınFITC Fare Anti-İnsan PAC-1</td><td>BD</td><td>340507</td><td></td></tr><tr><td>FITC-IETD-FMK Paz 8 (aktif) boyama kiti</td><td>Abcam</td><td>ab65614</td><td></td></tr><tr><td>Mitotracker Kırmızı CMXRos (mitochindria etiketleme boyası)</td><td>Invitrogen</td><td>M7512</td><td>Stok= 1 mM (50 & mikro; g 90 & mikro'da çözülmüş; l DMSO) Alt stok = 100 &mikro; M (10 & mikro; l Stok + 90 ve mikro; l DMSO) Çalışma konsantrasyonu = 500nM (0.5 & mikro; 100'de L ve mikro; l)</td></tr> <tr><td>PE Annexin V</td><td>BD Pharmingen</td><td>560930</td><td></td></tr><tr><td>Prostaglandin E1</td><td>Sigma</td><td>P5515</td><td>Stok = 20 mM (1 mg 141 & mikro; L DMSO) Alt stok= 1 mM (10 &mikro; l Stok + 190 & L DMSO) Çalışma konsantrasyonu= 3 &mikro; M (3 & 1 mL'de L)</td></tr><tr><td>Rhod-2</td><td>Invitrogen</td><td>R1244</td><td>Stok= 5 mM (178 & mikro'da çözünmüş 1 mg; L DMSO) Alt stok= 100 &mikro; M (10 & mikro; L Stok + 90 & L DMSO) Çalışma konsantrasyonu= 500 nM (0.5 &mikro; 100'de L ve mikro; L)</td></tr> <tr><td>İnsan plazmasından trombin</td><td>Sigma</td><td>T7572</td><td></td></tr></tbody></table>

nekroz, apoptoz ve trombosit aktivasyonu Özellikleri sergileyen prokoagülan trombositlerin kapsamlı analizi

Comprehensive Analysis of Procoagulant Platelets Exhibiting Features of Necrosis, Apoptosis and Platelet Activation

Aktive edilmiş trombositlerin bir kısmı, hem fosfatidilserin (PS) hem de P-selektin'in yüzey ekspresyonunda karakteristik bir artışla "prokoagülan" hale gelir, bu da onları sadece PS maruziyeti için pozitif olan "apoptotik" trombositlerden ve P-selektin ekspresyonu için pozitif olan "proagregatör" trombositlerden ayırır. Trombinin, sırasıyla FITC-anti-CD62P antikoru ve PE-Annexin V'nin bağlanmasıyla tespit edildiği gibi, hem P-selektin hem de PS ekspresyonu için pozitif olan prokoagülan trombositlerin oranında doza bağlı...

Nekroz, Apoptoz ve Trombosit Aktivasyonu Özellikleri Sergileyen Prokoagülan Trombositlerin Kapsamlı Analizi

Platelets circulating in the bloodstream are relatively quiescent but become "activated" upon encountering stimulants or "agonists" at the site of blood ...

comprehensive-analysis-procoagulant-platelets-exhibiting-features

Research

JoVE Journal

Medicine

Tıp

Burada vasküler inflamasyon ve canlı farelerde trombüs oluşumu sırasında aktive endotel üzerindeki heterotipik trombosit-nötrofil etkileşimleri görselleştirmek için floresan intravital mikroskobu deneysel bir tekniktir rapor. Bu mikroskobik teknoloji vasküler hastalığın moleküler mekanizmasını incelemek ve patofizyolojik durumlardaki farmakolojik ajanları test için değerli olacaktır.

Canlı Fare Damar İltihabı ve trombüs oluşumu sırasında Activated Endotel üzerinde heterotypic Trombosit nötrofil Etkileşimleri Gerçek zamanlı Görüntüleme

Bu çalışma, trombosit yapışması, yayılması ve akış altında salgı çalışması için bir floresan mikroskopi tabanlı yöntem açıklanmaktadır. Bu çok yönlü platform tromboz ve hemostaz üzerine mekanik araştırma için trombosit fonksiyonunun araştırılmasını sağlar.

Akım altında Trombosit Degranülasyonu ve Salgılamanın Canlı Hücreli Görüntüleme

Prokoagülan trombosit oluşumu, artmış tromboz riski ile ilişkilendirilmiştir. Burada, prokoagülan trombositlerin ayırt edici özellikleri olan fosfatidilserin ve mikrovezikül salınımının maruziyetini ölçmeyi amaçlayan, yıkanmış trombositleri insan kanından izole etmek için kesin bir protokol sunulmaktadır.

İnsan Saflaştırılmış Trombositlerinden Fosfatidilserin Maruziyeti ve Mikrovezikül Salınımının Ölçülmesiyle Prokoagülan Trombosit Karakterizasyonu

Akış altındaki trombosit fonksiyonu değerlendirilebilir ve simüle edilmiş hemostatik resüsitasyon, travma ve transfüzyon tıbbında uygulamaları olan bir mikroakışkan cihaz kullanılarak modellenebilir.

Trombosit Fonksiyonunun Değerlendirilmesinde Mikroakışkanlar

Trombositler için çok renkli akış sitometrisi, klasik trombosit alt popülasyonları içinde istirahat, agregatör, prokoagülan ve apoptotik trombositler gibi yeni trombosit alt tiplerini tanımlayabilir. Bu, çeşitli trombosit agonistleri tarafından indüklenen farklı alt popülasyon modellerinin karşılaştırılmasına izin verir. Burada, çok renkli bir akış sitometrisi paneli oluşturmak için bir prosedür ayrıntılı olarak açıklanmaktadır.

Trombosit Alt Popülasyonlarının Çok Renkli Akış Sitometrisi ile Analiz Edilmesi

Bu çalışma, kanda trombus oluşumunu tetiklemek için kesme stresi kullanan ve trombü birden fazla boyutlu okuma ile karakterize eden bir mikrofluidik testi tanımlar. Bu test, kan örneklerinin protrombotik eğilimini ölçebilir ve bu nedenle hastalık teşhisi, ilaç keşfi ve trombozla ilgili temel mekanik çalışmalarda faydalıdır.

Trombus Profilleme Testi: Biyomekanik Trombogenezin Kapsamlı Karakterizasyonu İçin Mikrofluidik Tabanlı Bir Platform

Torasik onkoloji veritabanı translasyonel araştırma amaçlı klinik ve laboratuvar veriler için kapsamlı bir veri havuzu olarak hizmet etmek için geliştirilmiştir. Veritabanı Göğüs Onkoloji Araştırma Programı çerçevesinde translasyonel kanser araştırmacıları hizmet edecektir. Bu veritabanı, diğer kanser modellerinin yanı sıra diğer insan hastalıkları için uyarlanabilir.

Kapsamlı Göğüs Onkoloji Veritabanı Üretimi - Translasyonel Araştırma Aracı

Konuşma desteği yol fizyolojik mekanizmaların Amaç değerlendirmeler ALS olan kişilerde hastalığın oluşumu ve ilerlemesinde izlemek ve klinik çalışmalarda tedavi etkilerini ölçmek için ihtiyaç vardır. Bu video, kapsamlı bir klinik popülasyonlarda konuşma motor performansının ölçülmesi için, enstrümantasyon tabanlı bir protokol mevcut.

Amyotrophic Lateral Sclerosis bulber Disfonksiyon Kapsamlı Değerlendirmesi (ALS) için Protokol

Kanser Hücreleri Endotel Hücre Tek tabakalı Invasion Tedbir Gerçek zamanlı Elektrik Empedans Tabanlı Tekniği

RNA müdahalesi (RNAi) gen nakavt üzerinden pek çok avantajı sahip ve geniş gen işlevsel çalışmalarda bir araç olarak kullanılmıştır. DNA vektör tabanlı RNAi teknolojisi buluşu uzun vadeli ve olası indüklenebilir gen Knockdown yaptı, hem de gen susturulması fizibilite...

Kanser gen Fonksiyon Eğitim için DNA vektör tabanlı RNA Girişim

Öğrenin, prokoagülan trombositleri nekroz, apoptoz ve aktivasyon özellikleriyle analiz etme yöntemini. Bu protokol, tromboz riski değerlendirmesi ve tedavi hedefleri için metabolik yolakları incelemeye yardımcı olur.

Nekroz, Apoptoz ve Trombosit Aktivasyonu Özellikleri Sergileyen Prokoagülan Trombositlerin Kapsamlı Analizi

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