Name: highly multiplexed, super-resolution imaging of t cells using madstorm
Uploaded: 2017-06-24T12:30:00
Description: Watch this Scientific Journal Video about Highly Multiplexed, Super-resolution Imaging of T Cells Using madSTORM at JoVE.com

Ringraziamo Xufeng Wu per l&#39;accesso al microscopio STORM. Questa ricerca è stata sostenuta dal Programma Intramurale di Ricerca del National Cancer Institute (NCI) Centro per la Ricerca sul Cancro e dal National Heart Lung and Blood Institute (NHLBI).

Attenzione: Prima dell&#39;utilizzo, consultare tutti i fogli di sicurezza relativi ai materiali di sicurezza (MSDS). Molte delle sostanze chimiche utilizzate in questo protocollo sono tossiche e cancerogene. Utilizzare tutte le appropriate pratiche di sicurezza durante l&#39;esecuzione del protocollo, compresa l&#39;uso di controlli ingegneristici (cappa di fumo, custodia di guanto) e dispositivi di protezione individuale (occhiali di protezione, guanti, cappotto di laboratorio, pantaloni lunghi, scarpe chiuse). <p...

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A., Zhuang, X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stochastic+optical+reconstruction+microscopy+(STORM):+a+method+for+superresolution+fluorescence+imaging.">Stochastic optical reconstruction microscopy (STORM): a method for superresolution fluorescence imaging.</a> Cold Spring Harb Protoc. (6), 498-520 (2013).</li><li>Yi, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=madSTORM:+a+superresolution+technique+for+large-scale+multiplexing+at+single-molecule+accuracy.">madSTORM: a superresolution technique for large-scale multiplexing at single-molecule accuracy.</a> Mol Biol Cell. 27 (22), 3591-3600 (2016).</li><li>Bates, M., Huang, B., Dempsey, G. T., Zhuang, X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Multicolor+super-resolution+imaging+with+photo-switchable+fluorescent+probes.">Multicolor super-resolution imaging with photo-switchable fluorescent probes.</a> Science. 317 (5845), 1749-1753 (2007).</li><li>Dempsey, G. T., Vaughan, J. C., Chen, K. 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A., Borbely, J. S., Sandoval Alvarez, A., Lakadamyali, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cross-talk-free+multi-color+STORM+imaging+using+a+single+fluorophore.">Cross-talk-free multi-color STORM imaging using a single fluorophore.</a> PLoS One. 9 (7), e101772 (2014).</li><li>Valley, C. C., Liu, S., Lidke, D. S., Lidke, K. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sequential+superresolution+imaging+of+multiple+targets+using+a+single+fluorophore.">Sequential superresolution imaging of multiple targets using a single fluorophore.</a> PLoS One. 10 (4), e0123941 (2015).</li><li>Hoebe, R. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Controlled+light-exposure+microscopy+reduces+photobleaching+and+phototoxicity+in+fluorescence+live-cell+imaging.">Controlled light-exposure microscopy reduces photobleaching and phototoxicity in fluorescence live-cell imaging.</a> Nat Biotechnol. 25 (2), 249-253 (2007).</li><li>Kiuchi, T., Higuchi, M., Takamura, A., Maruoka, M., Watanabe, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Multitarget+super-resolution+microscopy+with+high-density+labeling+by+exchangeable+probes.">Multitarget super-resolution microscopy with high-density labeling by exchangeable probes.</a> Nat Methods. 12 (8), 743-746 (2015).</li><li>Campi, G., Varma, R., Dustin, M. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Actin+and+agonist+MHC-peptide+complex-dependent+T+cell+receptor+microclusters+as+scaffolds+for+signaling.">Actin and agonist MHC-peptide complex-dependent T cell receptor microclusters as scaffolds for signaling.</a> J Exp Med. 202 (8), 1031-1036 (2005).</li><li>Sarkar, S. K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Wide-field+in+vivo+background+free+imaging+by+selective+magnetic+modulation+of+nanodiamond+fluorescence.">Wide-field in vivo background free imaging by selective magnetic modulation of nanodiamond fluorescence.</a> Biomed Opt Express. 5 (4), 1190-1202 (2014).</li></ol>

Le procedure di multiplexing sequenziali, di correzione della deriva e di allineamento in madSTORM consentono una visualizzazione precisa e altamente multipleplazionale delle strutture eterogenee nelle cellule. 10 Inoltre, madSTORM evita le limitazioni di STORM multicolore, come l&#39;aberrazione cromatica e le proprietà di scambio fotovoltaico / emissioni sub-ottimali di coloranti rossi non lontani 9 , 12 . Poiché la fase di eluizione ...

Sono state sviluppate varie tecniche di microscopia super-risoluzione per superare il limite di diffrazione della microscopia leggera (~ 200 nm). Tra queste è una categoria di tecniche chiamate microscopia di localizzazione singola (SMLM) che comprende microscopia di localizzazione foto-attivazione (PALM) e microscopia ottica stoccastica di ricostruzione (STORM). Le tecniche SMLM condividono l&#39;uso di fluorofori che possono essere commutati tra stati (fluorescenti) e fuori (scuri / foto-commutati), consentendo la localizzazione sequenziale della fluorescenza da singole molecole 1 , 2 , 3 . Grazie alla sua compatibilità con i coloranti e microscopi commercialmente disponibili, STORM diretto (dSTORM) è diventato una tecnica SMLM ampiamente adottata 4 . DSTORM riesce a raggiungere normalmente una precisione di localizzazione ~ 10 nm, definita come l&#39;incertezza nel calcolo del centro di un diffractFunzione di diffusione a punti ioni-limitata (PSF). Tuttavia, nonostante l&#39;alta precisione stimata utilizzando gli algoritmi di localizzazione 5 , 6 , 7 , un&#39;accurata determinazione della posizione effettiva delle singole molecole è stata ostacolata da una serie di problemi. Innanzitutto, il movimento meccanico dello stadio microscopico durante l&#39;acquisizione di immagini aggiunge un&#39;incertezza significativa alla precisione di localizzazione. Poiché le immagini SMLM sono ottenute in migliaia di frame di scadenza, i movimenti nano-scala dello stadio microscopico possono compromettere in modo significativo la precisione dell&#39;immagine finale super-risoluzione 8 . Per compensare il movimento della fase durante l&#39;acquisizione dell&#39;immagine, la deriva di fase è comunemente stimata dalla raccolta a regressione di localizzazioni binned dall&#39;immagine stessa (correlazione incrociata) o localizzazioni sequenziali da marcatori fiduciali (correzione fiduciaria) 1 , 9 . CarnQuesti metodi richiedono l&#39;ottimizzazione di più parametri per ogni stack di immagini e non possono rappresentare i movimenti di fase in scala temporanea come la vibrazione meccanica. Nano-particelle d&#39;oro e perline fluorescenti multicolori sono state utilizzate come marcatori fiduciali in SMLM ma non sono foto-stabili e risultano in una precisione notevolmente inferiore dopo la correzione della deriva rispetto ai nano-diamanti fluorescenti (FND) In madSTORM 10 . Oltre al limite di diffrazione, la microscopia leggera è ulteriormente limitata dai limiti spettrali. La visualizzazione simultanea di bersagli multipli richiede sonde fluorescenti con profili spettrali non sovrapposti, generalmente limitando la microscopia leggera a fluorescenza a 6 colori e SMLM a 2-3 colori 4 , 11 , 12 . Inoltre, l&#39;aberrazione cromatica non lineare provoca un disallineamento di immagini multicolor, wChe richiedono procedure di allineamento estese utilizzando marcatori fiduciali multicolori 8 , 13 . Per ovviare a questi limiti, studi precedenti hanno ripreso obiettivi multipli con fotopolimerizzazione ripetuta o spegnimento chimico dei fluorofori sequenzialmente legati 14 , 15 , 16 , 17 , 18 , 19 . Mentre questi metodi possono superare i limiti spettrali della microscopia, il sbiancamento di fluorescenza è noto per essere un processo tossico 20 e uno sbiancamento prolungato o uno spegnimento può causare effetti collaterali indesiderati, come la perdita di reticolazione. Inoltre, l&#39;accumulo di sonde fluorescenti potrebbe portare a blocchi sterici di siti di legame nel campione, impedendo la multiplexing su larga scala e il targeting robusto di epitopi. Per evitare tali interferenze steriche, un recente sHa ottenuto la multiplexing utilizzando uno scambio stocastico di frammenti proteici liberamente diffusi 21 . Mentre questo metodo consente una densa etichettatura delle strutture cellulari, richiede un&#39;ampia preparazione biochimica per isolare i frammenti peptidici, non individuare posizioni singole molecole e non agevolmente agevolare il multiplexing su larga scala utilizzando sonde commercialmente disponibili. Presentiamo un protocollo video dettagliato che descrive il legame sequenziale e l&#39;eluzione di anticorpi fluorescenti per l&#39;imaging dSTORM (madSTORM) multiplexato con dimensioni corporee e l&#39;utilizzo di nano diamanti fluorescenti per ottenere una corretta correzione e allineamento della deriva.

<table border="0" cellpadding="0" cellspacing="0" width="598">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:269px;">8 well coverslip chamber</td>
			<td style="width:117px;">Lab-tek</td>
			<td style="width:124px;">155409</td>
			<td style="width:88px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">0.1% Poly-L-Lysine solution</td>
			<td>Sigma-Aldrich</td>
			<td>P8920</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">NV-100nm Fluorescent Nano-diamond</td>
			<td>Adamas</td>
			<td>Red FND</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">anti-CD3&#949; antibody</td>
			<td>BD Biosciences</td>
			<td>555329</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Bovine serum albumin, Fraction V</td>
			<td>KSE Scientific</td>
			<td>98-100P</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Triton-X</td>
			<td>Sigma-Aldrich</td>
			<td>T9284</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">10% Paraformaldehyde</td>
			<td>EMS</td>
			<td>15712</td>
			<td>Carcinogen</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Gelatin from fresh water fish skin</td>
			<td>Sigma-Aldrich</td>
			<td>G7041</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Alexa 647 antibody labeling kit</td>
			<td>Thermo Fisher</td>
			<td>A20186</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Magnesium Chloride hexahydrate</td>
			<td>Sigma-Aldrich</td>
			<td>138924</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">PIPES</td>
			<td>Sigma-Aldrich</td>
			<td>P6757</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Tween-20</td>
			<td>Fisher Scientific</td>
			<td>BP337-500</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">2-Mercaptoethanol</td>
			<td>Sigma-Aldrich</td>
			<td>M7154</td>
			<td>Highly toxic, air sensitive</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Cysteamine</td>
			<td>Sigma-Aldrich</td>
			<td>30070</td>
			<td>Highly toxic&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Cyclooctatetraene 98%</td>
			<td>Sigma-Aldrich</td>
			<td>138924</td>
			<td>Highly toxic, air sensitive</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">10x PBS</td>
			<td>KD Medical</td>
			<td>RGF-3210</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">10x TBS</td>
			<td>KD Medical</td>
			<td>RGF-3385</td>
			<td></td>
		</tr>
	</tbody>
</table>

highly multiplexed, super-resolution imaging of t cells using madstorm

Imaging di strutture cellulari eterogenee utilizzando la microscopia di localizzazione a singola molecola è stata ostacolata da inadeguate capacità di localizzazione e di multiplexing. Utilizzando marcatori fiduciari a fluorescenza nano-diamante, descriviamo le procedure di correzione e di allineamento necessarie per ottenere un&#39;elevata precisione nella microscopia di localizzazione singola molecola. Inoltre, viene descritta una nuova strategia di multiplexing, madSTORM, in cui molecole multiple sono mirate nella stessa cellula usando sequenziali e eluendo degli anticorpi fluorescenti. MadSTORM è dimostrato su una cellula T attivata per visualizzare le posizioni di diversi componenti all&#39;interno di una struttura a più proteine ​​legata alla membrana chiamata microcluster del recettore cellulare T. Inoltre, viene discussa l&#39;applicazione di madSTORM come strumento generale per la visualizzazione di strutture multi-proteine.

L&#39;eluizione sequenziale e il metodo di colorazione sono stati utilizzati per produrre l&#39;immagine madstorm multiplex di microclustri e altre strutture in una cella di Jurkat T attivata ( Figura 1 , allineamenti della cifra di controllo). Ogni immagine pseudo-colorata rappresenta un ciclo di imaging madSTORM acquisito nei passaggi da 1.1.1 a 1.3.13. Come descritto in precedenza 10 , il campo visivo dovrebbe essere monitorato per...

Watch this Scientific Journal Video about Highly Multiplexed, Super-resolution Imaging of T Cells Using madSTORM at JoVE.com

Highly Multiplexed, Super-resolution Imaging of T Cells Using madSTORM

Abbiamo dimostrato un metodo per l&#39;immagine di molteplici molecole all&#39;interno di nano-strutture eterogenee alla precisione di una singola molecola utilizzando il legame sequenziale e l&#39;eluzione di anticorpi a fluorescenza.

Molto multiplo, la risoluzione super-risoluzione delle celle T utilizzando la madSTORM

Imaging heterogeneous cellular structures using single molecule localization microscopy has been hindered by inadequate localization precision and ...

The overall goal of this imaging protocol is to visualize multiple proteins in an activated T cell, using single molecule localisation microscopy. This method can help answer key questions in the T cell activation field such as, the single molecule distribution of TC or microcluster components. The main advantage of this technique is that, it allows greater multiplexing than achieved by current supervised solution microscopy methods.To begin, directly conjugate antibodies with Alexa 647 dye, using the Alexa 647 antibody labeling kit, following the labeling protocol provided by the manufacturer. Centrifuge the labeled antibodies for five minutes at 20, 800 times G.Following centrifugation, collect the supernatant to remove the aggregated antibodies. Next, coat A12 cover slip chambers with 250 microliters of 01%poly-l-lyzine for 15 minutes.Then, aspirate the solution without rinsing and dry the chambers at 65 degree celsius for 30 minutes. Prepare dilution of 100 nanometer fluorescent nanodiamonds or FNDs, in one PBS, test various dilutions to ensure that enough FNDs will be visible in each field. Vortex the diluted FNDs for one minute.Next, sonicate the FND supernatant for 30 seconds at a high power setting. Incubate the sonicated FND supernatant in a poly-l-lyzine coated A12 cover slip chamber for 30 minutes at room temperature. After washing the chamber five times, with 1x PBS, visualize the FND coated chamber using 647 nanometer laser excitation on the turf microscope.Use a 100x objective tealed a 61 by 61 micron field of view. Ideally, four to 10 individual FNDs should be visible in the field of view, given the appropriate dilution of FNDs, with at least one FND present in each quadrant of the imaging field. Once the proper density of FNDs has been achieved, add 250 microliters of anti-CD3 antibody to each well.Incubate the chamber for one hour at 37 degree celsius or overnight at four degree celsius. Following incubation, remove the solution from the wells and add 1x PBS for 30 seconds. Repeat this wash step five times.Spin one milliliter of Jurkat T cells at 800 times G for six minutes. Jurkat T cells should ideally be at a concentration of 0.5 to one million cells per milliliter concentration before spinning. Re-suspend the cells in 300 microliters of 1x HBS solution.Meanwhile, add 150 microliters of 1x HBS solution to each well of the chamber and incubate at 37 degree celsius for 15 minutes. Then, add 50 microliters of the re-suspended Jurkat T cells to each well and incubate for three minutes at 37 degree celsius. Next, add 300 microliters of 4%paraformaldehyde to each well and incubate for 30 minutes at 37 degree celsius.After washing three times with 1x PBS, permeabilise the cells, by adding 250 microliters of 01%Triton x solution for five minutes at room temperature. Following three additional washes with 1x PBS, add 250 microliters of 1%fish gelatin solution to each well for 30 minutes at room temperature. Finally, wash the cells three more times with 1x PBS.Add 200 microliters of labeled antibody at 0.1 to 0.5 micrograms per milliliter to the fixed cells for one hour at room temperature. After washing the cells five times with 1x PBS, add one milliliter of STORM buffer and cover the chamber with a glass cover slip to limit exposure. Use clamps on the microscope stage to hold the cover slip chamber in position.This step is critical for large scale multiplexing, since the same field of view must be present after several rounds of washing and incubation. Using a low 647 nanometer laser power in turf mode, locate a stained cell with at least three FNDs in the field of view. Increase the 647 nanometer laser power and acquire the images.Typical parameters can be found in the text protocol. Wash the cells five times with 1x tris-buffered saline or 1x TBS. Then, add one milliliter of elution buffer and incubate at room temperature for one minute.Repeat the elution three times. Use the exact elution conditions and a number of elution rinses needed to remove the signal as specified for each antibody. Then, wash the cells three times with 1x TBS and add one milliliter of 1x PBS.Proper execution of this elution step, will result in removal of previously bound antibody and allow higher efficiency for subsequent antibody labeling. It is important to use TBS for washes to minimize precipitation between elution steps. Photo bleach the cells using a 647 nanometer laser at high power, with a 405 nanometer laser at two to five milliwatts to photo activate Alexa 647 dyes in the dark state.Wait until all remaining signal from non-eluted antibody is photo bleached. Typically, this takes two to five seconds of laser exposure. Acquire sample images, using the STORM settings after illusion and photo bleaching, to confirm removal of the signal.Next, add 250 microliters of 4%paraformaldehyde for 15 minutes. This prevents reverse cross-linking of fixed molecules in the cell. Wash the cells three times with 1x PBS.Repeat these steps for sequential labeling of multiple targets. Finally, perform drift correction and alignment of multiplex image stacks at described in the text protocol. The sequential illusion and standing method was used to produce the multiplex MET STORM image of microclusters and other structures in an activated Jurkat T cell.The final MET STORM image has been corrected for a drift and alignment, using average fiducial correction and FND fiducial markers. Shown here, are single molecule localisation microscopy images of a single FND localized in 30, 000 image frames before and after drift correction with average fiducial correction. Similarly, here are single molecule localisation microscopy images of an activated Jurkat T cell stained with anti-phosphorelated SLP76 antibody and FND futusial markers, before and after drift correction with average futusial correction.After watching this video, you should have a good understanding of how to perform multiplexed super resolution imaging of proteins in an activated T cell, using MET STORM. Once mastered, this technique can be done in three hours for each multiplexing round. While attempting this procedure, it's important to remember to keep the specimen in the same position on the microscope stage.This can be done using stage clamps. Don't forget that working with our modified STORM buffer, can be extremely hazardous and precautions such as, respirators and goggles should always be taken while performing this procedure.

Research

JoVE Journal

Immunology and Infection

Retroviral Transduction of T-cell Receptors in Mouse T-cells

Trasduzione retrovirale di recettori delle cellule T nel topo le cellule T

T-cell receptors (TCRs) play a central role in the immune system. TCRs on T-cell surfaces can specifically recognize peptide antigens presented by antigen ...

Ricerca

Immunologia e infezioni

Ex vivo Imaging of T Cells in Murine Lymph Node Slices with Widefield and Confocal Microscopes

Ex vivo Imaging of T Cells in Murine Lymph Node Slices with Widefield and Confocal Microscopes

Imaging ex vivo di cellule T nel Fette murini Linfonodo con grandangolari e microscopi confocale

Na&iuml;ve T cells continuously traffic to secondary lymphoid organs, including peripheral lymph nodes, to detect rare expressed antigens. The migration ...

Expansion of Human Peripheral Blood  &gamma;&delta; T Cells using Zoledronate

Expansion of Human Peripheral Blood  &#947;&#948; T Cells using Zoledronate

Espansione del sangue periferico umano γδ cellule T con Zoledronato

Human &gamma;&delta; T cells can recognize and respond to a wide variety of stress-induced antigens, thereby developing innate broad anti-tumor and ...

Generation of Induced Regulatory T Cells from Primary Human Na&iuml;ve and Memory T Cells

Generation of Induced Regulatory T Cells from Primary Human Na&#239;ve and Memory T Cells

Generazione di cellule T regolatrici indotte da cellule primarie umane T naive e memoria

The development and maintenance of immunosuppressive CD4+ regulatory T cells (Tregs) contribute to the peripheral tolerance needed to remain in ...

Highly Resolved Intravital Striped-illumination Microscopy of Germinal Centers

Altamente Risolto intravitale Striped-illuminazione Microscopia dei Centri Germinal

Monitoring cellular communication by intravital deep-tissue multi-photon microscopy is the key for understanding the fate of immune cells within thick ...

Averaging of Viral Envelope Glycoprotein Spikes from Electron Cryotomography Reconstructions using Jsubtomo

Della media di Viral Busta Glicoproteina Spikes da Electron Cryotomography Ricostruzioni con Jsubtomo

Enveloped viruses utilize membrane glycoproteins on their surface to mediate entry into host cells. Three-dimensional structural analysis of these ...

Super-resolution Imaging of the Natural Killer Cell Immunological Synapse on a Glass-supported Planar Lipid Bilayer

Super-Resolution Imaging di cellule Natural Killer immunologica Sinapsi su un vetro supportato Planar doppio strato lipidico

The glass-supported planar lipid bilayer system has been utilized in a variety of disciplines. One of the most useful applications of this technique has ...

Analysis of Yersinia enterocolitica Effector Translocation into Host Cells Using Beta-lactamase Effector Fusions

Analysis of Yersinia enterocolitica Effector Translocation into Host Cells Using Beta-lactamase Effector Fusions

Analisi di<em&gt; Yersinia enterocolitica</em&gt; Effector traslocazione in cellule ospiti Utilizzando beta-lattamasi Effector Fusions

Many gram-negative bacteria including pathogenic Yersinia spp. employ type III secretion systems to translocate effector proteins into eukaryotic target ...

Cortical Actin Flow in T Cells Quantified by Spatio-temporal Image Correlation Spectroscopy of Structured Illumination Microscopy Data

Corticale actina di flusso in cellule T quantificati da spazio-temporale Correlazione Immagine Spettroscopia di Structured Illuminazione Microscopia dati

Filamentous-actin plays a crucial role in a majority of cell processes including motility and, in immune cells, the formation of a key cell-cell ...

In Vitro Differentiation of Human CD4+FOXP3+ Induced Regulatory T Cells (iTregs) from Na&#239;ve CD4+ T Cells Using a TGF-&#946;-containing Protocol

La differenziazione in vitro di cellule CD4 + umana FOXP3 + indotta da T regolatorie (iTregs) da cellule T CD4 Naïve utilizzando un protocollo TGF-β contenenti