Wir danken Prof. Ferenc Krausz für die Diskussionen und Najd Altwaijry für ihre Unterstützung für die Fertigstellung des Manuskripts. Diese Arbeit wurde durch das Center for Advanced Laser Applications (CALA) finanziert.

<ol>
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C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Terahertz+emission+profile+from+laser-induced+air+plasma.">Terahertz emission profile from laser-induced air plasma.</a> Appl Phys Lett. 88 (26), 261103 (2006).</li><li>Herrmann, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Generation+of+sub-three-cycle,+16+TW+light+pulses+by+using+noncollinear+optical+parametric+chirped-pulse+amplification.">Generation of sub-three-cycle, 16 TW light pulses by using noncollinear optical parametric chirped-pulse amplification.</a> Opt Lett. 34 (16), 2459-2461 (2009).</li><li>Adachi, S., 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=1.5+mJ,+6.4+fs+parametric+chirped-pulse+amplification+system+at+1+kHz.">1.5 mJ, 6.4 fs parametric chirped-pulse amplification system at 1 kHz.</a> Opt Lett. 32 (17), 2487-2489 (2007).</li><li>Adachi, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=5-fs,+multi-mJ,+CEP-locked+parametric+chirped-pulse+amplifier+pumped+by+a+450-nm+source+at+1+kHz.">5-fs, multi-mJ, CEP-locked parametric chirped-pulse amplifier pumped by a 450-nm source at 1 kHz.</a> Opt express. 16 (19), 14341-14352 (2008).</li><li>Yin, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=High-efficiency+optical+parametric+chirped-pulse+amplifier+in+BiB3O6+for+generation+of+3+mJ,+two-cycle,+carrier-envelope-phase-stable+pulses+at+1.7+&#181;m.">High-efficiency optical parametric chirped-pulse amplifier in BiB3O6 for generation of 3 mJ, two-cycle, carrier-envelope-phase-stable pulses at 1.7 &#181;m.</a> Opt Lett. 41 (6), 1142-1145 (2016).</li><li>Deng, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Carrier-envelope-phase-stable,+1.2+mJ,+1.5+cycle+laser+pulses+at+2.1+&#181;m.">Carrier-envelope-phase-stable, 1.2 mJ, 1.5 cycle laser pulses at 2.1 &#181;m.</a> Opt Lett. 37 (23), 4973-4975 (2012).</li><li>Rothhardt, J., Demmler, S., H&#228;drich, S., Limpert, J., T&#252;nnermann, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Octave-spanning+OPCPA+system+delivering+CEP-stable+few-cycle+pulses+and+22+W+of+average+power+at+1+MHz+repetition+rate.">Octave-spanning OPCPA system delivering CEP-stable few-cycle pulses and 22 W of average power at 1 MHz repetition rate.</a> Opt express. 20 (10), 10870-10878 (2012).</li><li>Heckl, O. H., Nolte, S., Schrempel, F., Dausinger, F., 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=.">.</a> Ultrafast Thin-Disk Lasers.Ultrashort Pulse Laser Technology. 195, 93-115 (2016).</li><li>Zapata, L. E., 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=Cryogenic+Yb:YAG+composite-thin-disk+for+high+energy+and+average+power+amplifiers.">Cryogenic Yb:YAG composite-thin-disk for high energy and average power amplifiers.</a> Opt. Lett. 40 (11), 2610-2613 (2015).</li><li>Schulz, M., 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=Yb:YAG+Innoslab+amplifier:+efficient+high+repetition+rate+subpicosecond+pumping+system+for+optical+parametric+chirped+pulse+amplification.">Yb:YAG Innoslab amplifier: efficient high repetition rate subpicosecond pumping system for optical parametric chirped pulse amplification.</a> Opt Lett. 36 (13), 2456-2458 (2011).</li><li>Roeser, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Millijoule+pulse+energy+high+repetition+rate+femtosecond+fiber+chirped-pulse+amplification+system.">Millijoule pulse energy high repetition rate femtosecond fiber chirped-pulse amplification system.</a> Opt Lett. 32 (24), 3495-3497 (2007).</li><li>Russbueldt, P., 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=400+W+Yb:YAG+Innoslab+fs-amplifier.">400 W Yb:YAG Innoslab fs-amplifier.</a> Opt Express. 17 (15), 12230-12245 (2009).</li><li>Baumgarten, C., 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=0.5+kHz+repetition+rate+picosecond+laser.">0.5 kHz repetition rate picosecond laser.</a> Opt Lett. 41 (14), 3339-3342 (2016).</li><li>Klingebiel, S., 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=220mJ,+1+kHz+Picosecond+Regenerative+Thin-Disk+Amplifier.">220mJ, 1 kHz Picosecond Regenerative Thin-Disk Amplifier.</a> , (2015).</li><li>Nubbemeyer, T., 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=1+kW,+200+mJ+picosecond+thin-disk+laser+system.">1 kW, 200 mJ picosecond thin-disk laser system.</a> Opt Lett. 42 (7), 1381-1384 (2017).</li><li>Fattahi, H., 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=High-power,+1-ps,+all-Yb:YAG+thin-disk+regenerative+amplifier.">High-power, 1-ps, all-Yb:YAG thin-disk regenerative amplifier.</a> Opt Lett. 41 (6), 1126-1129 (2016).</li><li>Brons, 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=Energy+scaling+of+Kerr-lens+mode-locked+thin-disk+oscillators.">Energy scaling of Kerr-lens mode-locked thin-disk oscillators.</a> Opt Lett. 39 (22), 6442-6445 (2014).</li><li>Trebino, R., 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=Measuring+ultrashort+laser+pulses+in+the+time-frequency+domain+using+frequency-resolved+optical+gating.">Measuring ultrashort laser pulses in the time-frequency domain using frequency-resolved optical gating.</a> Rev Sci Instrum. 68 (9), 3277-3295 (1997).</li><li>Arisholm, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=General+numerical+methods+for+simulating+second-order+nonlinear+interactions+in+birefringent+media.">General numerical methods for simulating second-order nonlinear interactions in birefringent media.</a> J Opt Soc Am B. 14 (10), 2543-2549 (1997).</li><li>Zhang, D. 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Die Autoren haben nichts zu offenbaren.

Der schlüsselfertige Betrieb des Oszillators wird durch die optimale Wärmemanagement der verschiedenen Komponenten des Lasers erreicht. Der Ausgang des Oszillators ist täglich reproduzierbar, ohne dass eine zusätzliche Ausrichtung oder Optimierung erforderlich ist. Darüber hinaus erfüllt die Puls-zu-Puls-Energiestabilität und die räumliche Punktstabilität des Saatlasers die Voraussetzungen, um den stabilen Betrieb des regenerativen Verstärkers zu erreichen. Andere Niedrigenergie-Saatquellen, wie z. B. Faserverstärker, können verwendet werden, um den Verstärker zu säen. In dieser Studie wurde ein 2 μJ Yb: YAG-Dünnschicht-KLM-Oszillator verwendet, um die Verstärkung des regenerativen Verstärkers durch Verringerung des Wachstums der angesammelten nichtlinearen Phasen zu unterstützen, da die erforderliche Anzahl von Rundfahrten für höher eingeführte Samenergie verringert wird . Darüber hinaus beeinflusst die höhere Saatgut-Energie den Verstärkungsprozess und reduziert die Verstärkung. Die gemessene spektrale Bandbreite der verstärkten ImpulseEs für verschiedene Samenergien bei einer festen Pumpenleistung ist in Abbildung 5c ​​gezeigt . Verstärkte spektrale Bandbreite sinkt für niedrigere Samenergien wegen der Verringerung der Verstärkung. Für 10 pJ Samenenergie arbeitet der Laser in der Periodenverdopplung, und es ist nicht möglich, einen stabilen Betrieb zu erreichen, auch wenn die Anzahl der Rundfahrten erhöht wird. Neben der sorgfältigen Optimierung der Kühlsysteme und der Stromversorgung der Dioden spielt der Betrieb des regenerativen Verstärkers bei der Sättigung eine wesentliche Rolle bei der erreichten Stabilität des Verstärkers. Die grundlegende oder zweite Harmonische des Lasers kann verwendet werden, um ein OPCPA-System zu pumpen. Für SHG wurden die Leistungen eines LBO und eines BBO-Kristalls verglichen, da sie trotz des größeren räumlichen Walk-offs und der begrenzten verfügbaren Blende im Falle von BBO einen hohen nichtlinearen Koeffizienten und Schadensschwelle bieten. Da der nichtlineare Koeffizient von BBO fast doppelt so hoch ist wie der LBO, ist ein kürzerer Kristall sufUm die Sättigungsgrenze für SHG zu erreichen ( Abbildung 6a ). Daher ist BBO die geeignetere Wahl, da die akkumulierte nichtlineare Phase kleiner ist 28 . Die Pulsdauern der SH-Impulse werden experimentell bei unterschiedlichen Umwandlungswirkungsgraden charakterisiert. Es wurde beobachtet, dass bei hohen Umwandlungswirkungsgraden das SHG-Spektrum erweitert und eine höherwertige Spektralphase erscheint ( Abbildung 6 ). Daher wird der Fall B mit dem Umwandlungswirkungsgrad von 70% gewählt, wo der SH und die nicht umgesetzten Grundstrahlen eine ausgezeichnete Qualität erhalten.

Die Erzeugung von hochenergetischen, wenigen Zyklus-Laserpulsen mit einer hohen Wiederholrate ist für angewandte Felder wie die Attosekundenwissenschaft 1 , 2 , 3 , 4 und die Hochfeldphysik 5 , 6 von großem Interesse, die direkt zugute kommen Aus der Verfügbarkeit solcher Quellen. OPCPA stellt den vielversprechendsten Weg zur Erzielung hoher Pulsenergien und großer Verstärkungsbandbreiten dar, die gleichzeitig wenige Zyklusimpulse unterstützen 1 . Bis heute ermöglicht OPCPA eine Ultrabreitbandverstärkung, die wenige Zyklusimpulse 7 , 8 , 9 , 10 erzeugt . Eine modifizierte Implementierung des OPCPA-Schemas, das kurze Pumpimpulse auf der Pikosekunden-Skala verwendet, hält jedoch VersprechenWobei dieser Ansatz für noch höhere Puls-Energien und durchschnittliche Potenzen im wenigen Zyklus-Regime 1 , 11 , 12 skalierbar ist. Aufgrund der hohen Pumpenintensität bei kurzimpulsgepumptem OPCPA ermöglicht die hohe Single-Pass-Verstärkung die Verwendung von sehr dünnen Kristallen, um große Verstärkungsbandbreiten zu unterstützen. Obwohl das kurzpulsgepumpte OPCPA viele Vorteile hat, unterliegt die Realisierbarkeit dieses Ansatzes der Verfügbarkeit von Lasern, die speziell für diesen Zweck zugeschnitten sind. Solche Pumplaser sind verpflichtet, hochenergetische Pikosekundenimpulse mit nahezu beugungsbegrenzter Strahlqualität bei Wiederholraten im kHz bis MHz Bereich 13 , 14 , 15 zu liefern. Die Einführung von Ytterbium-dotierten Lasern in verschiedenen Geometrien, die in der Lage sind, Pikosekunden-Laserpulse mit hoher Energie und hoher Durchschnittsleistung zu liefernUm den aktuellen Zustand des Feldes 1 , 13 , 14 , 15 , 16 , 17 , 18 zu ändern. Yb: YAG hat eine gute Wärmeleitfähigkeit und eine lange Lebensdauer des oberen Lebens und kann durch kostengünstige Diodenlaser gepumpt werden. Seine Leistung bei Verwendung in Dünnschicht-Geometrie ist hervorragend aufgrund der effizienten Kühlung des Verstärkungsmediums, um gleichzeitig die Spitzen- und Durchschnittsleistung zu skalieren. Darüber hinaus wird das Auftreten von Selbstfokussierung innerhalb des Verstärkungsmediums während des Verstärkungsprozesses aufgrund der Schlankheit der Dünnschicht im Vergleich zu anderen Verstärkungsmediumgeometrien unterdrückt, was zu exzellenten zeitlichen und räumlichen Profilen der verstärkten Impulse führt. Die Kombination dieses Konzepts mit CPA hält Versprechen für die Erzeugung von Pikosekunden-Impulsen mit Hunderten von Millajoulen Energie und HundertenVon Watt der durchschnittlichen Macht 19 , 20 . Ziel dieser Arbeit ist es, einen schlüsselfertigen Yb: YAG-Dünnschicht-Regenerationsverstärker mit hervorragender Tagesleistung als geeignete Quelle für das Pumpen von OPCPAs 21 zu demonstrieren. Um dieses Ziel zu erreichen, verwendet diese Studie einen Yb: YAG-Dünnschicht-Oszillator 22 mit mehreren Mikrokoppeln von Pulsenergie, um den Verstärker zu säubern, um die akkumulierte nichtlineare Phase während des Verstärkungsprozesses zu minimieren. Dieses Protokoll liefert das Rezept für den Bau und Betrieb des Lasersystems, das an anderer Stelle beschrieben wird 21 . Details zur Komponentenimplementierung und Steuerungssoftware werden vorgestellt und der Ausrichtungsprozess des Systems beschrieben.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Electrooptics</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Fiber-Coupled Diode Laser Module</td>
			<td>Dilas Diodenlaser GmbH</td>
			<td>M1F8H12-940.5-500C-IS11.34</td>
			<td></td>
		</tr>
		<tr>
			<td>Fiber-Coupled Diode Laser Module</td>
			<td>Laserline GmbH</td>
			<td>LDM1000-500</td>
			<td></td>
		</tr>
		<tr>
			<td>Power Supply for Diode Laser</td>
			<td>Delta Elektronika B.V.</td>
			<td>SM 15-100</td>
			<td></td>
		</tr>
		<tr>
			<td>Power Supply for Diode Laser</td>
			<td>Delta Elektronika B.V.</td>
			<td>SM 35-45</td>
			<td></td>
		</tr>
		<tr>
			<td>Pulse Picker&#39;s Driver</td>
			<td>Bergmann Messger&#228;te Entwicklung KG&#160;</td>
			<td>N/A, customized</td>
			<td></td>
		</tr>
		<tr>
			<td>Pockels Cell&#39;s Driver</td>
			<td>Bergmann Messger&#228;te Entwicklung KG&#160;</td>
			<td>N/A, customized</td>
			<td></td>
		</tr>
		<tr>
			<td>Pulse Picker&#39;s Driver Power Supply</td>
			<td>Bergmann Messger&#228;te Entwicklung KG&#160;</td>
			<td>PCD8m7</td>
			<td></td>
		</tr>
		<tr>
			<td>Pockels Cell&#39;s Driver Power Supply</td>
			<td>Bergmann Messger&#228;te Entwicklung KG&#160;</td>
			<td>PCD8m7</td>
			<td></td>
		</tr>
		<tr>
			<td>Delay Generator PCI</td>
			<td>Bergmann Messger&#228;te Entwicklung KG&#160;</td>
			<td>BME_SG08p</td>
			<td></td>
		</tr>
		<tr>
			<td>Splitter Box</td>
			<td>Bergmann Messger&#228;te Entwicklung KG&#160;</td>
			<td>N/A, customized</td>
			<td></td>
		</tr>
		<tr>
			<td>Resonant Preamplifier</td>
			<td>Bergmann Messger&#228;te Entwicklung KG&#160;</td>
			<td>BME_P03</td>
			<td></td>
		</tr>
		<tr>
			<td>Pulse Picker&#39;s crystal</td>
			<td>Castech Inc.</td>
			<td>N/A, customized</td>
			<td>12*12*20 mm&#179;</td>
		</tr>
		<tr>
			<td>Pockels Cell&#39;s crystal</td>
			<td>Castech Inc.</td>
			<td>N/A, customized</td>
			<td>12*12*20 mm&#179;</td>
		</tr>
		<tr>
			<td>Name</td>
			<td>Company</td>
			<td>Catalog Number</td>
			<td>Comments</td>
		</tr>
		<tr>
			<td>Optics</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Thin-disk</td>
			<td>TRUMPF Scientific Lasers&#160;</td>
			<td>N/A, customized</td>
			<td></td>
		</tr>
		<tr>
			<td>Thin-disk Head</td>
			<td>TRUMPF Scientific Lasers&#160;</td>
			<td>N/A, customized</td>
			<td></td>
		</tr>
		<tr>
			<td>Fiber</td>
			<td>Frank Optic Products GmbH</td>
			<td>N/A, customized</td>
			<td></td>
		</tr>
		<tr>
			<td>Fiber Objective</td>
			<td>Edmund Optics GmbH</td>
			<td>N/A, customized</td>
			<td></td>
		</tr>
		<tr>
			<td>Faraday Isolator</td>
			<td>Electro-Optics Technology, Inc</td>
			<td>EOT.189.12231</td>
			<td></td>
		</tr>
		<tr>
			<td>Faraday Rotator</td>
			<td>Electro-Optics Technology, Inc</td>
			<td>EOT.189.22040</td>
			<td></td>
		</tr>
		<tr>
			<td>Stretcher&#39;s Grating 1</td>
			<td>Horiba Jobin Yvon GmbH</td>
			<td>N/A, customized</td>
			<td>60*40*10 mm&#179;</td>
		</tr>
		<tr>
			<td>Stretcher&#39;s Grating 2</td>
			<td>Horiba Jobin Yvon GmbH</td>
			<td>N/A, customized</td>
			<td>350*190*50 mm&#179;</td>
		</tr>
		<tr>
			<td>Compressor&#39;s Grating 1</td>
			<td>Plymouth Grating Laboratory, Inc.</td>
			<td>N/A, customized</td>
			<td>40*40*16 mm&#179;</td>
		</tr>
		<tr>
			<td>Compressor&#39;s Grating 2</td>
			<td>Plymouth Grating Laboratory, Inc.</td>
			<td>N/A, customized</td>
			<td>300*100*50 mm&#179;</td>
		</tr>
		<tr>
			<td>HR Mirror, 1&#34; (1030nm), flat, 0&#176;</td>
			<td>Layertec GmbH</td>
			<td>108060</td>
			<td></td>
		</tr>
		<tr>
			<td>HR Mirror, 1&#34; (1030nm), flat, 0&#176;</td>
			<td>Laseroptik GmbH</td>
			<td>B-09965, S-04484</td>
			<td></td>
		</tr>
		<tr>
			<td>HR Mirror, 1&#34; (1030nm), flat, 45&#176;</td>
			<td>Layertec GmbH</td>
			<td>108063</td>
			<td></td>
		</tr>
		<tr>
			<td>HR Mirror, 1&#34; (1030nm), flat, 45&#176;</td>
			<td>Laseroptik GmbH</td>
			<td>B-09966, S-04484</td>
			<td></td>
		</tr>
		<tr>
			<td>HR Mirror, 1&#34; (1030nm), curved</td>
			<td>Layertec GmbH</td>
			<td>N/A, customized</td>
			<td>set</td>
		</tr>
		<tr>
			<td>HR Mirror, 2&#34; (1030nm), flat, 0&#176;</td>
			<td>Laseroptik GmbH</td>
			<td>B-09965, S-05474</td>
			<td></td>
		</tr>
		<tr>
			<td>HR Mirror, 2&#34; (1030nm), flat, 45&#176;</td>
			<td>Laseroptik GmbH</td>
			<td>B-09966, S-05474</td>
			<td></td>
		</tr>
		<tr>
			<td>Thin Film Polarizer (1030nm), 2&#34;</td>
			<td>Layertec GmbH</td>
			<td>103930</td>
			<td></td>
		</tr>
		<tr>
			<td>Waveplate L/2 (1030nm)</td>
			<td>Layertec GmbH</td>
			<td>106058</td>
			<td>&#216;=25mm</td>
		</tr>
		<tr>
			<td>Waveplate L/4 (1030nm)</td>
			<td>Layertec GmbH</td>
			<td>106060</td>
			<td>&#216;=25mm</td>
		</tr>
		<tr>
			<td>AR Window (1030nm), wedge</td>
			<td>Laseroptik GmbH</td>
			<td>B-00183-01, S-00988</td>
			<td>&#216;=38mm</td>
		</tr>
		<tr>
			<td>Output Coupler, 1&#34; (1030nm)</td>
			<td>Layertec GmbH</td>
			<td>N/A, customized</td>
			<td>PR = 88 %</td>
		</tr>
		<tr>
			<td>High-dispersion Mirror (1030nm)</td>
			<td>UltraFast Innovations GmbH</td>
			<td>N/A, customized</td>
			<td>GDD = -3000 fs&#178;</td>
		</tr>
		<tr>
			<td>Lens, 1&#34; (1030nm), Plano-Convex</td>
			<td>Thorlabs GmbH</td>
			<td>N/A, customized</td>
			<td>set</td>
		</tr>
		<tr>
			<td>Lens, 1&#34; (1030nm), Plano-Concave</td>
			<td>Thorlabs GmbH</td>
			<td>N/A, customized</td>
			<td>set</td>
		</tr>
		<tr>
			<td>Lens, 2&#34; (1030nm), Plano-Convex</td>
			<td>Thorlabs GmbH</td>
			<td>N/A, customized</td>
			<td>set</td>
		</tr>
		<tr>
			<td>Lens, 2&#34; (1030nm), Plano-Concave</td>
			<td>Thorlabs GmbH</td>
			<td>N/A, customized</td>
			<td>set</td>
		</tr>
		<tr>
			<td>HR Mirror, 1&#34; (515nm), flat, 0&#176;</td>
			<td>Layertec GmbH</td>
			<td>129784</td>
			<td></td>
		</tr>
		<tr>
			<td>HR Mirror, 1&#34; (515nm), flat, 0&#176;</td>
			<td>Eksma Optics</td>
			<td>042-0515-i0</td>
			<td></td>
		</tr>
		<tr>
			<td>HR Mirror, 1&#34; (515nm), flat, 45&#176;</td>
			<td>Layertec GmbH</td>
			<td>110924</td>
			<td></td>
		</tr>
		<tr>
			<td>HR Mirror, 1&#34; (515nm), flat, 45&#176;</td>
			<td>Eksma Optics</td>
			<td>042-0515</td>
			<td></td>
		</tr>
		<tr>
			<td>HR Mirror, 1&#34; (515nm), curved</td>
			<td>Layertec GmbH</td>
			<td>N/A, customized</td>
			<td>set</td>
		</tr>
		<tr>
			<td>HR Mirror, 1&#34; (515nm), curved</td>
			<td>Eksma Optics</td>
			<td>N/A, customized</td>
			<td>set</td>
		</tr>
		<tr>
			<td>HR Mirror, 2&#34; (515nm), flat, 0&#176;</td>
			<td>Eksma Optics</td>
			<td>045-0515-i0</td>
			<td></td>
		</tr>
		<tr>
			<td>HR Mirror, 2&#34; (515nm), flat, 45&#176;</td>
			<td>Eksma Optics</td>
			<td>045-0515</td>
			<td></td>
		</tr>
		<tr>
			<td>Thin Film Polarizer (515nm), 2&#34;</td>
			<td>Layertec GmbH</td>
			<td>112544</td>
			<td></td>
		</tr>
		<tr>
			<td>Waveplate L/2 (515nm)</td>
			<td>Layertec GmbH</td>
			<td>112546</td>
			<td>&#216;=25mm</td>
		</tr>
		<tr>
			<td>Lens, 1&#34; (515nm), Plano-Convex</td>
			<td>Thorlabs GmbH</td>
			<td>N/A, customized</td>
			<td>set</td>
		</tr>
		<tr>
			<td>Lens, 1&#34; (515nm), Plano-Concave</td>
			<td>Thorlabs GmbH</td>
			<td>N/A, customized</td>
			<td>set</td>
		</tr>
		<tr>
			<td>Kerr Medium</td>
			<td>Meller Optics, Inc.</td>
			<td>N/A, customized</td>
			<td>Sapphire, 1mm</td>
		</tr>
		<tr>
			<td>BBO Crystal</td>
			<td>Castech Inc.</td>
			<td>N/A, customized</td>
			<td>7*7*1.5 mm&#179;</td>
		</tr>
		<tr>
			<td>Harmonic Separator, 1&#34;, 45&#176;</td>
			<td>Eksma Optics</td>
			<td>042-5135</td>
			<td></td>
		</tr>
		<tr>
			<td>Harmonic Separator, 2&#34;, 45&#176;</td>
			<td>Eksma Optics</td>
			<td>045-5135</td>
			<td></td>
		</tr>
		<tr>
			<td>Silver Mirror, 1&#34;, flat</td>
			<td>Thorlabs GmbH</td>
			<td>PF10-03-P01</td>
			<td></td>
		</tr>
		<tr>
			<td>Silver Mirror, 1&#34;, curved</td>
			<td>Eksma Optics</td>
			<td>N/A, customized</td>
			<td>set</td>
		</tr>
		<tr>
			<td>Filter - Absorptive Neutral Density</td>
			<td>Thorlabs GmbH</td>
			<td>NE##A</td>
			<td>set</td>
		</tr>
		<tr>
			<td>Filter - Reflective Neutral Density</td>
			<td>Thorlabs GmbH</td>
			<td>ND##A</td>
			<td>set</td>
		</tr>
		<tr>
			<td>Filter - Round Continuously Variable</td>
			<td>Thorlabs GmbH</td>
			<td>NDC-50C-4M</td>
			<td></td>
		</tr>
		<tr>
			<td>Filter - Edgepass Filter (Longpass)</td>
			<td>Thorlabs GmbH</td>
			<td>FEL####</td>
			<td>set</td>
		</tr>
		<tr>
			<td>Filter - Edgepass Filter (Shortpass)</td>
			<td>Thorlabs GmbH</td>
			<td>FES####</td>
			<td>set</td>
		</tr>
		<tr>
			<td>Wedge</td>
			<td>Thorlabs GmbH</td>
			<td>N/A, customized</td>
			<td>set</td>
		</tr>
		<tr>
			<td>Name</td>
			<td>Company</td>
			<td>Catalog Number</td>
			<td>Comments</td>
		</tr>
		<tr>
			<td>Optomechanics &#38; Motion</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Mirror Mount 1&#34; (small)</td>
			<td>S. Maier GmbH</td>
			<td>S1M4-##-1&#8221;</td>
			<td></td>
		</tr>
		<tr>
			<td>Mirror Mount 1&#34; (large)</td>
			<td>S. Maier GmbH</td>
			<td>S3-##</td>
			<td></td>
		</tr>
		<tr>
			<td>Mirror Mount 1&#34;</td>
			<td>TRUMPF Scientific Lasers&#160;</td>
			<td>1&#34; adjustable&#160;</td>
			<td></td>
		</tr>
		<tr>
			<td>Mirror Mount 2&#34;</td>
			<td>S. Maier GmbH</td>
			<td>S4-##</td>
			<td></td>
		</tr>
		<tr>
			<td>Mirror Mount 2&#34;</td>
			<td>TRUMPF Scientific Lasers&#160;</td>
			<td>2&#34; adjustable&#160;</td>
			<td></td>
		</tr>
		<tr>
			<td>Rotation Mount 1&#8221;</td>
			<td>S. Maier GmbH</td>
			<td>D25</td>
			<td></td>
		</tr>
		<tr>
			<td>Rotation Mount 1&#8221;</td>
			<td>Thorlabs GmbH</td>
			<td>RSP1/M</td>
			<td></td>
		</tr>
		<tr>
			<td>Rotation Mount 2&#8221;</td>
			<td>Thorlabs GmbH</td>
			<td>RSP2/M</td>
			<td></td>
		</tr>
		<tr>
			<td>Precision Rotation Stage</td>
			<td>Newport Corporation</td>
			<td>M-UTR120</td>
			<td></td>
		</tr>
		<tr>
			<td>Four-Axis Diffraction Grating Mount</td>
			<td>Newport Corporation</td>
			<td>DGM-1</td>
			<td></td>
		</tr>
		<tr>
			<td>Translation Stage</td>
			<td>OptoSigma Corporation</td>
			<td>TADC-651SR25-M6</td>
			<td></td>
		</tr>
		<tr>
			<td>Pockels cell stage</td>
			<td>Newport Corporation</td>
			<td>9082-M</td>
			<td></td>
		</tr>
		<tr>
			<td>Pockels Cell Holder</td>
			<td>Home-made</td>
			<td>N/A, customized</td>
			<td></td>
		</tr>
		<tr>
			<td>Picomotor Controller/Driver Kit</td>
			<td>Newport Corporation</td>
			<td>8742-12-KIT</td>
			<td></td>
		</tr>
		<tr>
			<td>Picomotor Piezo Linear Actuators</td>
			<td>Newport Corporation</td>
			<td>8301NF</td>
			<td></td>
		</tr>
		<tr>
			<td>Picomotor Rotation Mount</td>
			<td>Newport Corporation</td>
			<td>8401-M</td>
			<td></td>
		</tr>
		<tr>
			<td>Hand Control Pad</td>
			<td>Newport Corporation</td>
			<td>8758</td>
			<td></td>
		</tr>
		<tr>
			<td>Name</td>
			<td>Company</td>
			<td>Catalog Number</td>
			<td>Comments</td>
		</tr>
		<tr>
			<td>Light Analysis</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Beam Profiling Camera</td>
			<td>Ophir Optronics Solutions Ltd</td>
			<td>SP620</td>
			<td></td>
		</tr>
		<tr>
			<td>Beam Profiling Camera</td>
			<td>DataRay Inc.</td>
			<td>WCD-UCD23</td>
			<td></td>
		</tr>
		<tr>
			<td>Photodiodes (solw)</td>
			<td>Thorlabs GmbH</td>
			<td>DET10A/M</td>
			<td></td>
		</tr>
		<tr>
			<td>Photodiodes (fast)</td>
			<td>Alphalas GmbH</td>
			<td>UPD-200-SP</td>
			<td></td>
		</tr>
		<tr>
			<td>Thin-disk Camera</td>
			<td>Imaging Development Systems GmbH</td>
			<td>UI-2220SE-M-GL</td>
			<td></td>
		</tr>
		<tr>
			<td>Oscilloscope</td>
			<td>Tektronix GmbH</td>
			<td>DPO5204</td>
			<td></td>
		</tr>
		<tr>
			<td>Oscilloscope</td>
			<td>Teledyne LeCroy GmbH</td>
			<td>SDA 760Zi-A</td>
			<td></td>
		</tr>
		<tr>
			<td>Spectrometer</td>
			<td>Avantes</td>
			<td>AvaSpec-ULS3648-USB2</td>
			<td></td>
		</tr>
		<tr>
			<td>Spectrometer</td>
			<td>Ocean Optics, Inc</td>
			<td>HR4C1769</td>
			<td></td>
		</tr>
		<tr>
			<td>Spectrometer</td>
			<td>Ocean Optics, Inc</td>
			<td>HR4C3762</td>
			<td></td>
		</tr>
		<tr>
			<td>Spectrometer</td>
			<td>Ocean Optics, Inc</td>
			<td>HR4D464</td>
			<td></td>
		</tr>
		<tr>
			<td>Spectrometer</td>
			<td>Ocean Optics, Inc</td>
			<td>HR4D466</td>
			<td></td>
		</tr>
		<tr>
			<td>Laser Thermal Power Sensor</td>
			<td>Ophir Optronics Solutions Ltd</td>
			<td>L50(150)A-PF-35</td>
			<td></td>
		</tr>
		<tr>
			<td>Laser Thermal Power Sensor</td>
			<td>Ophir Optronics Solutions Ltd</td>
			<td>FL500A</td>
			<td></td>
		</tr>
		<tr>
			<td>Laser Thermal Power Sensor</td>
			<td>Ophir Optronics Solutions Ltd</td>
			<td>3A-P-V1</td>
			<td></td>
		</tr>
		<tr>
			<td>Power and Energy Meter</td>
			<td>Ophir Optronics Solutions Ltd</td>
			<td>Vega</td>
			<td></td>
		</tr>
		<tr>
			<td>Name</td>
			<td>Company</td>
			<td>Catalog Number</td>
			<td>Comments</td>
		</tr>
		<tr>
			<td>Systems</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Laser Beam Stabilization System</td>
			<td>TEM-Messtechnik GmbH</td>
			<td>Aligna</td>
			<td></td>
		</tr>
		<tr>
			<td>Laser M&#178; Measuring System</td>
			<td>Ophir Optronics Solutions Ltd</td>
			<td>M&#178;-200s</td>
			<td></td>
		</tr>
		<tr>
			<td>FROG</td>
			<td>Home-made</td>
			<td>N/A, customized</td>
			<td></td>
		</tr>
		<tr>
			<td>XFROG</td>
			<td>Home-made</td>
			<td>N/A, customized</td>
			<td></td>
		</tr>
		<tr>
			<td>Name</td>
			<td>Company</td>
			<td>Catalog Number</td>
			<td>Comments</td>
		</tr>
		<tr>
			<td>Miscellaneous</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Cooling Chiller</td>
			<td>H.I.B Systemtechnik GmbH</td>
			<td>6HE-000800-W-W-R23-2-DI</td>
			<td></td>
		</tr>
		<tr>
			<td>Cooling Chiller</td>
			<td>Termotek GmbH</td>
			<td>P201</td>
			<td></td>
		</tr>
		<tr>
			<td>Cooling Chiller</td>
			<td>Termotek GmbH</td>
			<td>P208</td>
			<td></td>
		</tr>
		<tr>
			<td>Laser Safety Goggles</td>
			<td>Protect - Laserschutz GmbH</td>
			<td>BGU 10-0165-G-20</td>
			<td></td>
		</tr>
		<tr>
			<td>Infra-red Viewer</td>
			<td>FJW Optical Systems</td>
			<td>84499A</td>
			<td></td>
		</tr>
		<tr>
			<td>Laser Viewing Card</td>
			<td>Thorlabs GmbH</td>
			<td>VRC4</td>
			<td></td>
		</tr>
		<tr>
			<td>Laser Viewing Card</td>
			<td>Thorlabs GmbH</td>
			<td>VRC5</td>
			<td></td>
		</tr>
		<tr>
			<td>Laser Viewing Card</td>
			<td>Laser Components GmbH</td>
			<td>LDT-1064 BG</td>
			<td></td>
		</tr>
		<tr>
			<td>Flowmeter</td>
			<td>KOBOLD Messring GmbH</td>
			<td>DTK-1250G2C34P</td>
			<td></td>
		</tr>
		<tr>
			<td>Pressure Gauge</td>
			<td>KOBOLD Messring GmbH</td>
			<td>EN 837-1</td>
			<td></td>
		</tr>
		<tr>
			<td>Temperature Sensor</td>
			<td>KOBOLD Messring GmbH</td>
			<td>TDA-15H* ***P3M</td>
			<td></td>
		</tr>
		<tr>
			<td>WinLase Software</td>
			<td>Dr. C. Horvath &#38; Dr. F. Loesel</td>
			<td>WinLase Version 2.1 pro.</td>
			<td>Laser Cavity Software</td>
		</tr>
	</tbody>
</table>

20 mj, 1 ps yb:yag thin-disk regenerative amplifier

Dies ist ein Bericht über einen 100 W, 20 mJ, 1 ps Yb: YAG Dünnschicht-regenerative Verstärker. Ein hausgemachter Yb: YAG-Dünnschicht-, Kerr-Objektiv-Modus-gesperrter Oszillator mit schlüsselfertiger Leistung und Mikrojoule-Puls-Energie wird verwendet, um den regenerativen Chirped-Puls-Verstärker zu säen. Der Verstärker befindet sich im luftdichten Gehäuse. Es arbeitet bei Raumtemperatur und zeigt einen stabilen Betrieb bei einer Wiederholrate von 5 kHz bei einer Puls-zu-Puls-Stabilität von weniger als 1%. Durch die Verwendung eines 1,5 mm dicken Beta-Barium-Borat-Kristalls wird die Frequenz der Laserleistung auf 515 nm verdoppelt, mit einer durchschnittlichen Leistung von 70 W, was einem optisch-optischen Wirkungsgrad von 70% entspricht. Diese überlegene Leistung macht das System zu einer attraktiven Pumpquelle für optische parametrische Chirped-Puls-Verstärker im Nah-Infrarot- und Mid-Infrarot-Spektralbereich. Durch die Kombination der schlüsselfertigen Leistung und der überlegenen Stabilität des regenerativen Verstärkers erleichtert das System die Erzeugung eines breitbandigen, CEP-stabilenSamen. Die Bereitstellung des Saatguts und der Pumpe der optischen parametrischen Chirped-Puls-Verstärkung (OPCPA) von einer Laserquelle eliminiert den Bedarf einer aktiven zeitlichen Synchronisation zwischen diesen Impulsen. Diese Arbeit stellt eine detaillierte Anleitung zum Aufbau und Betrieb eines Yb: YAG-Dünnschicht-Regenerationsverstärkers, basierend auf Chirped-Puls-Verstärkung (CPA), als Pumpquelle für einen optischen parametrischen Chirped-Puls-Verstärker dar.

<html> Der Oszillator liefert 350 fs, 2 μJ, 25-W-Impulse bei 11 MHz Wiederholrate mit einer Puls-zu-Puls-Stabilität von 1% (rms) und Strahlpunktschwankungen von weniger als 0,6% über 1 h Messwert ( Abbildung 4 ). <img alt="Abbildung 4" class="xfigimg" src="/files/ftp_upload/55717/55717fig4.jpg" /> Abbildung 4 : Yb: YAG-Dünnschicht, Kerr-Linsen-Modus-gesperrter Oszillator. ( A ) Das Spektrum (rot), das abgerufene zeitliche Intensitätsprofil (blau) und das räumliche Profil (Einfügung) der Oszillatorimpulse. ( B ) Gemessener und abgerufener SHG-FROG-Spektrograph des Oszillators. Diese Figur wurde von Fattahi et al. , Mit Erlaubnis aus Referenz 21 .&gt; Bitte klicken Sie hier, um eine größere Version dieser Figur zu sehen. Die Seed-Impulse werden im regenerativen Verstärker auf 125 W verstärkt, während sie mit einer CW-fasergekoppelten Diode bei einer Wellenlänge von 940 nm bei 280 W gepumpt werden, was einer optisch-optischen Effizienz von 47% entspricht. Die Puls-zu-Puls-Stabilität des Verstärkers beträgt weniger als 1%, und der Verstärker weist nach 10 h Dauerbetrieb eine ausgezeichnete Langzeitstabilität auf. Der verstärkte Strahl hat ein ausgezeichnetes Raumprofil mit einem M 2 von 1 (M 2 x = 1,08 und M 2 y = 1,07) und einem ausgezeichneten zeitlichen Profil nach Kompression auf 1 ps (bei FWHM) ( Abbildung 5 ). <img alt="Abbildung 5" class="xfigimg" src="/files/ftp_upload/55717/55717fig5.jpg" /> Abbildung 5 : Charakterisierung des regenerativen VerstärkersAusgabe und der Verstärkungsverengung. ( A ) Die Stabilität der regenerativen Verstärker-Durchschnittsleistung nach 10 h Dauerbetrieb. Inset: ( a-1 ) Normalisierte Leistung auf ihren Mittelwert in einem Zeitfenster von 0,5 h; ( A-2 ) Ausgangsstrahlprofil des regenerativen Verstärkers. ( B ) Verstärkerausgangsspektrum (grün) und die abgerufene zeitliche Intensität (blau) der Laserpulse bei 100 W durchschnittlicher Leistung nach dem Gitterverdichter. ( C ) Samenergie gegen Spektralbandbreite (FWHM) des Verstärkerausgangs und die erforderlichen Rundfahrten für die gleiche Ausgangsleistung bei 300 W Pumpleistung. Diese Figur wurde von Fattahi et al. , Mit Erlaubnis aus Referenz 21 . <a href="http://ecsource.jove.com/files/ftp_upload/55717/55717fig5large.jpg" target="_blank">Bitte klicken Sie hier, um eine größere Version dieser Figur zu sehen.</a> 25 analysiert. Es wurden zwei verschiedene Kristalle mit folgenden Parametern in Betracht gezogen: 1) ein Typ-I, 6 mm dickes Lithiumtriborat (LBO) mit einem Phasenanpassungswinkel von 13,7 ° und einem nichtlinearen Koeffizienten von 0,819 pm / V und 2) a Typ-I, 3 mm dickes BBO mit einem Phasenanpassungswinkel von 23,4 ° und einem nichtlinearen Koeffizienten von 2 pm / V 26 , 27 . 1-ps, 20-mJ-Pulse bei 1.030 nm und eine Peak-Intensität von 100 GW / cm 2 wurden als Eingang der Simulation betrachtet. Die Simulationsergebnisse zeigten, dass die BBO-Leistung dem des LBO für SHG überlegen war ( Abbildung 6 ). <img alt="Abbildung 6" class="xfigimg" src="/files/ftp_upload/55717/55717fig6.jpg" /> Abbildung 6 : Zweite Harmonische Generation. ( A ) simuliertes SHG eneRgy für einen 6 mm dicken LBO-Kristall und einen 3 mm dicken BBO-Kristall. ( B ) Experimentelle SHG-Effizienz gegenüber Input-Inputity-Intensität in einem 1,5 mm dicken BBO-Kristall mit 0,5 mJ (schwarz) und 20 mJ (grün) des Verstärkerausgangs. ( C ) Die abgetastete spektrale Intensität und ( d ) die Gruppenverzögerung von XFROG-Messungen für verschiedene SHG-Effizienzen entsprechend den Punkten A, B und C in (b). Diese Figur wurde von Fattahi et al. , Mit Erlaubnis aus Referenz 21 . <a href="http://ecsource.jove.com/files/ftp_upload/55717/55717fig6large.jpg" target="_blank">Bitte klicken Sie hier, um eine größere Version dieser Figur zu sehen.</a>

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20 mJ, 1 ps Yb:YAG Thin-disk Regenerative Amplifier

Ein Protokoll für den Betrieb einer hochenergetischen, hochleistungsfähigen, optisch parametrischen Chirp-Impulsverstärker-Pumpquelle auf der Basis eines Yb: YAG-Dünnschicht-Regenerationsverstärkers ist hier dargestellt.

20 mJ, 1 ps Yb: YAG Dünnschicht-Regenerationsverstärker

This is a report on a 100 W, 20 mJ, 1 ps Yb:YAG thin-disk regenerative amplifier. A homemade Yb:YAG thin-disk, Kerr-lens mode-locked oscillator with ...

20-mj-1-ps-ybyag-thin-disk-regenerative-amplifier

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11.4K Aufrufe.  Ludwig Maximilian University of Munich.  Ein Protokoll für den Betrieb einer hochenergetischen, hochleistungsfähigen, optisch parametrischen Chirp-Impulsverstärker-Pumpquelle auf der Basis eines Yb: YAG-Dünnschicht-Regenerationsverstärkers ist hier dargestellt. 

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Microfluidic Picoliter Bioreactor for Microbial Single-cell Analysis: Fabrication, System Setup, and Operation

In this protocol the fabrication, experimental setup and basic operation of the recently introduced microfluidic picoliter bioreactor (PLBR) is described in detail. The PLBR can be utilized for the analysis of single bacteria and microcolonies to investigate biotechnological and microbiological related questions concerning, e.g.&#160;cell growth, morphology, stress response, and metabolite or protein production on single-cell level. The device features continuous media flow enabling constant environmental conditions for perturbation studies, but in addition allows fast medium changes as well as oscillating conditions to mimic any desired environmental situation. To fabricate the single use devices, a silicon wafer containing sub micrometer sized SU-8 structures served as the replication mold for rapid polydimethylsiloxane casting. Chips were cut, assembled, connected, and set up onto a high resolution and fully automated microscope suited for time-lapse imaging, a powerful tool for spatio-temporal cell analysis. Here, the biotechnological platform organism Corynebacterium glutamicum&#160;was seeded into the PLBR and cell growth and intracellular fluorescence were followed over several hours unraveling time dependent population heterogeneity on single-cell level, not possible with conventional analysis methods such as flow cytometry. Besides insights into device fabrication, furthermore, the preparation of the preculture, loading, trapping of bacteria, and the PLBR cultivation of single cells and colonies is demonstrated. These devices will add a new dimension in microbiological research to analyze time dependent phenomena of single bacteria under tight environmental control. Due to the simple and relatively short fabrication process the technology can be easily adapted at any microfluidics lab and simply tailored towards specific needs.

In this protocol the fabrication, setup and basic operation of a microfluidic picoliter bioreactor (PLBR) for single-cell analysis of prokaryotic microorganisms is introduced. Industrially relevant microorganisms were analyzed as proof of principle allowing insights into growth rate, morphology, and phenotypic heterogeneity over certain time periods, hardly possible with conventional methods.

In this protocol the fabrication, experimental setup and basic operation of the recently introduced microfluidic picoliter bioreactor (PLBR) is described ...

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Harmonic Nanoparticles for Regenerative Research

In this visualized experiment, protocol details are provided for in vitro labeling of human embryonic stem cells (hESC) with second harmonic generation nanoparticles (HNPs). The latter are a new family of probes recently introduced for labeling biological samples for multi-photon imaging. HNPs are capable of doubling the frequency of excitation light by the nonlinear optical process of second harmonic generation with no restriction on the excitation wavelength.
Multi-photon based methodologies for hESC differentiation into cardiac clusters (maintained as long term air-liquid cultures) are presented in detail. In particular, evidence on how to maximize the intense second harmonic (SH) emission of isolated HNPs during 3D monitoring of beating cardiac tissue in 3D is shown. The analysis of the resulting images to retrieve 3D displacement patterns is also detailed.

Protocol details are provided for in vitro labeling human embryonic stem cells with second harmonic generating nanoparticles. Methodologies for hESC investigation by multi-photon microscopy and their differentiation into cardiac clusters are also presented.

Harmonic Nanopartikel für Regenerative Forschung

In this visualized experiment, protocol details are provided for in vitro labeling of human embryonic stem cells (hESC) with second harmonic generation ...

Generation of a Three-dimensional Full Thickness Skin Equivalent and Automated Wounding

In vitro models are a cost effective and ethical alternative to study cutaneous wound healing processes. Moreover, by using human cells, these models reflect the human wound situation better than animal models. Although two-dimensional models are widely used to investigate processes such as cellular migration and proliferation, models that are more complex are required to gain a deeper knowledge about wound healing. Besides a suitable model system, the generation of precise and reproducible wounds is crucial to ensure comparable results between different test runs. In this study, the generation of a three-dimensional full thickness skin equivalent to study wound healing is shown. The dermal part of the models is comprised of human dermal fibroblast embedded in a rat-tail collagen type I hydrogel. Following the inoculation with human epidermal keratinocytes and consequent culture at the air-liquid interface, a multilayered epidermis is formed on top of the models. To study the wound healing process, we additionally developed an automated wounding device, which generates standardized wounds in a sterile atmosphere.

The goal of this protocol is to build up a three-dimensional full thickness skin equivalent, which resembles natural skin. With a specifically constructed automated wounding device, precise and reproducible wounds can be generated under maintenance of sterility.

Erzeugung eines dreidimensionalen Vollhaut Equivalent und automatisierte Verwundung

In vitro models are a cost effective and ethical alternative to study cutaneous wound healing processes. Moreover, by using human cells, these models ...

Neonatal Cardiac Scaffolds: Novel Matrices for Regenerative Studies

The only definitive therapy for end stage heart failure is orthotopic heart transplantation. Each year, it is estimated that more than 100,000 donor hearts are needed for cardiac transplantation procedures in the United States1-2. Due to the limited numbers of donors, only approximately 2,400 transplants are performed each year in the U.S.2. Numerous approaches, from cell therapy studies to implantation of mechanical assist devices, have been undertaken, either alone or in combination, in an attempt to coax the heart to repair itself or to rest the failing heart3. In spite of these efforts, ventricular assist devices are still largely used for the purpose of bridging to transplantation and the utility of cell therapies, while they hold some curative promise, is still limited to clinical trials. Additionally, direct xenotransplantation has been attempted but success has been limited due to immune rejection. Clearly, another strategy is required to produce additional organs for transplantation and, ideally, these organs would be autologous so as to avoid the complications associated with rejection and lifetime immunosuppression. Decellularization is a process of removing resident cells from tissues to expose the native extracellular matrix (ECM) or scaffold. Perfusion decellularization offers complete preservation of the three dimensional structure of the tissue, while leaving the bulk of the mechanical properties of the tissue intact4. These scaffolds can be utilized for repopulation with healthy cells to generate research models and, possibly, much needed organs for transplantation. We have exposed the scaffolds from neonatal mice (P3), known to retain remarkable cardiac regenerative capabilities,5-8 to detergent mediated decellularization and we repopulated these scaffolds with murine cardiac cells. These studies support the feasibility of engineering a neonatal heart construct. They further allow for the investigation as to whether the ECM of early postnatal hearts may harbor cues that will result in improved recellularization strategies.

In these studies, we provide methodology for novel, neonatal, murine cardiac scaffolds for use in regenerative studies.

Neonatal Cardiac Scaffolds: Neuartige Matrices für Regenerative Studies

The only definitive therapy for end stage heart failure is orthotopic heart transplantation.  Each year, it is estimated that more than 100,000 donor ...

Studying Soft-matter and Biological Systems over a Wide Length-scale from Nanometer and Micrometer Sizes at the Small-angle Neutron Diffractometer KWS-2

The KWS-2 SANS diffractometer is dedicated to the investigation of soft matter and biophysical systems covering a wide length scale, from nm to &#181;m. The instrument is optimized for the exploration of the wide momentum transfer Q range between 1x10-4 and 0.5 &#197;-1 by combining classical pinhole, focusing (with lenses), and time-of-flight (with chopper) methods, while simultaneously providing high-neutron intensities with an adjustable resolution. Because of its ability to adjust the intensity and the resolution within wide limits during the experiment, combined with the possibility to equip specific sample environments and ancillary devices, the KWS-2 shows a high versatility in addressing the broad range of structural and morphological studies in the field. Equilibrium structures can be studied in static measurements, while dynamic and kinetic processes can be investigated over time scales between minutes to tens of milliseconds with time-resolved approaches. Typical systems that are investigated with the KWS-2 cover the range from complex, hierarchical systems that exhibit multiple structural levels (e.g., gels, networks, or macro-aggregates) to small and poorly-scattering systems (e.g., single polymers or proteins in solution). The recent upgrade of the detection system, which enables the detection of count rates in the MHz range, opens new opportunities to study even very small biological morphologies in buffer solution with weak scattering signals close to the buffer scattering level at high Q.
In this paper, we provide a protocol to investigate samples with characteristic size levels spanning a wide length scale and exhibiting ordering in the mesoscale structure using KWS-2. We present in detail how to use the multiple working modes that are offered by the instrument and the level of performance that is achieved.

Here, we present a protocol to investigate soft matter and biophysical systems over a wide mesoscopic length scale, from nm to &#181;m that involves the use of the KWS-2 SANS diffractometer at high intensities and an adjustable resolution.

Studieren weicher Materie und biologischer Systeme über eine Breite Länge-Skala von Nanometer- und Mikrometer Größen im Kleinwinkel Neutronendiffraktometer KWS-2

The KWS-2 SANS diffractometer is dedicated to the investigation of soft matter and biophysical systems covering a wide length scale, from nm to &#181;m. ...

3D Microtissues for Injectable Regenerative Therapy and High-throughput Drug Screening

To upgrade traditional 2D cell culture to 3D cell culture, we have integrated microfabrication with cryogelation technology to produce macroporous microscale cryogels (microcryogels), which can be loaded with a variety of cell types to form 3D microtissues. Herein, we present the protocol to fabricate versatile 3D microtissues and their applications in regenerative therapy and drug screening. Size and shape-controllable microcryogels can be fabricated on an array chip, which can be harvested off-chip as individual cell-loaded carriers for injectable regenerative therapy or be further assembled on-chip into 3D microtissue arrays for high-throughput drug screening. Due to the high elastic nature of these microscale cryogels, the 3D microtissues exhibit great injectability for minimally invasive cell therapy by protecting cells from mechanical shear force during injection. This ensures enhanced cell survival and therapeutic effect in the mouse limb ischemia model. Meanwhile, assembly of 3D microtissue arrays in a standard 384-multi-well format facilitates the use of common laboratory facilities and equipment, enabling high-throughput drug screening on this versatile 3D cell culture platform.

This protocol describes the fabrication of elastic 3D macroporous microcryogels by integrating microfabrication with cryogelation technology. Upon loading with cells, 3D microtissues are generated, which can be readily injected in vivo to facilitate regenerative therapy or assembled into arrays for in vitro high-throughput drug screening.

3D Microtissues für injizierbare Regenerative Therapie und Hochdurchsatz-Drogen-Screening

To upgrade traditional 2D cell culture to 3D cell culture, we have integrated microfabrication with cryogelation technology to produce macroporous ...

Generic Protocol for Optimization of Heterologous Protein Production Using Automated Microbioreactor Technology

A core business in industrial biotechnology using microbial production cell factories is the iterative process of strain engineering and optimization of bioprocess conditions. One important aspect is the improvement of cultivation medium to provide an optimal environment for microbial formation of the product of interest. It is well accepted that the media composition can dramatically influence overall bioprocess performance. Nutrition medium optimization is known to improve recombinant protein production with microbial systems and thus, this is a rewarding step in bioprocess development. However, very often standard media recipes are taken from literature, since tailor-made design of the cultivation medium is a tedious task that demands microbioreactor technology for sufficient cultivation throughput, fast product analytics, as well as support by lab robotics to enable reliability in liquid handling steps. Furthermore, advanced mathematical methods are required for rationally analyzing measurement data and efficiently designing parallel experiments such as to achieve optimal information content.
The generic nature of the presented protocol allows for easy adaption to different lab equipment, other expression hosts, and target proteins of interest, as well as further bioprocess parameters. Moreover, other optimization objectives like protein production rate, specific yield, or product quality can be chosen to fit the scope of other optimization studies. The applied Kriging Toolbox (KriKit) is a general tool for Design of Experiments (DOE) that contributes to improved holistic bioprocess optimization. It also supports multi-objective optimization which can be important in optimizing both upstream and downstream processes.

This manuscript describes a generic approach for tailor-made design of microbial cultivation media. This is enabled by an iterative workflow combining Kriging-based experimental design and microbioreactor technology for sufficient cultivation throughput, which is supported by lab robotics to increase reliability and speed in liquid handling media preparation.

Generischen Protokoll zur Optimierung der heterologen Proteinproduktion mit automatisierten Microbioreactor Technologie

A core business in industrial biotechnology using microbial production cell factories is the iterative process of strain engineering and optimization of ...

Recombinant Collagen I Peptide Microcarriers for Cell Expansion and Their Potential Use As Cell Delivery System in a Bioreactor Model

Tissue engineering is a promising field, focused on developing solutions for the increasing demand on tissues and organs regarding transplantation purposes. The process to generate such tissues is complex, and includes an appropriate combination of specific cell types, scaffolds, and physical or biochemical stimuli to guide cell growth and differentiation. Microcarriers represent an appealing tool to expand cells in a three-dimensional (3D) microenvironment, since they provide higher surface-to volume ratios and mimic more closely the in vivo situation compared to traditional two-dimensional methods. The vascular system, supplying oxygen and nutrients to the cells and ensuring waste removal, constitutes an important building block when generating engineered tissues. In fact, most constructs fail after being implanted due to lacking vascular support. In this study, we present a protocol for endothelial cell expansion on recombinant collagen-based microcarriers under dynamic conditions in spinner flask and bioreactors, and we explain how to determine in this setting cell viability and functionality. In addition, we propose a method for cell delivery for vascularization purposes without additional detachment steps necessary. Furthermore, we provide a strategy to evaluate the cell vascularization potential in a perfusion bioreactor on a decellularized&#160;biological matrix. We believe that the use of the presented methods could lead to the development of new cell-based therapies for a large range of tissue engineering applications in the clinical practice.

We propose a cell expansion protocol on macroporous microcarriers and their use as delivery system in a perfusion bioreactor to seed a decellularized tissue matrix. We also include different techniques to determine cell proliferation and viability of cells cultured on microcarriers. Furthermore, we demonstrate functionality of cells after bioreactor cultures.

Rekombinante Kollagen Modell I Peptid-Microcarriers für Zelle Expansion und ihre mögliche Verwendung als Zelle-Delivery-System in einem Bioreaktor

Tissue engineering is a promising field, focused on developing solutions for the increasing demand on tissues and organs regarding transplantation ...

Construction of a Multilayered Mesenchymal Stem Cell Sheet with a 3D Dynamic Culture System

Stem cell therapy shows a promising future in regenerating injured organ and tissues, and the cell sheet technique has been developed to improve the low cell retention and poor survival within the target zone. However, during the in vitro construction process, a solution for maintaining stem cell bioactivity and increasing the cell amount within the cell sheet is urgently needed. Here, this protocol presents a method for constructing a multilayered cell sheet with favorable stem cell bioactivity and optimal operability. Decellularized porcine pericardium (DPP) is prepared by phospholipase A2 (PLA2) decellularization method as the cell sheet scaffold, and rat bone marrow mesenchymal stem cells (BMSCs) are isolated and expanded as the seeded cells. The temporary multilayered cell sheet structure is constructed by using RAD16-I peptide hydrogel. Finally, the cell sheet is cultured with a dynamic perfusion system to stabilize the three-dimensional (3D) structure, and the cell sheet could be obtained following a 48-hour culture in vitro. This protocol provides an efficient and feasible method for constructing a multilayered stem cell sheet, and the cell sheet could be developed as a favorable stem cell therapy product in the future.

This article provides an efficient and feasible method for constructing multilayered stem cell sheets with favorable stem cell property.

Bau eines vielschichtigen mesenchymale Stammzellen-Blatt mit einem 3D dynamische Kultursystem

Stem cell therapy shows a promising future in regenerating injured organ and tissues, and the cell sheet technique has been developed to improve the low ...

Fabrication of the Composite Regenerative Peripheral Nerve Interface (C-RPNI) in the Adult Rat

Recent advances in neuroprosthetics have enabled those living with extremity loss to reproduce many functions native to the absent extremity, and this is often accomplished through integration with the peripheral nervous system. Unfortunately, methods currently employed are often associated with significant tissue damage which prevents prolonged use. Additionally, these devices often lack any meaningful degree of sensory feedback as their complex construction dampens any vibrations or other sensations a user may have previously depended on when using more simple prosthetics. The composite regenerative peripheral nerve interface (C-RPNI) was developed as a stable, biologic construct with the ability to amplify efferent motor nerve signals while providing simultaneous afferent sensory feedback. The C-RPNI consists of a segment of free dermal and muscle graft secured around a target mixed sensorimotor nerve, with preferential motor nerve reinnervation of the muscle graft and sensory nerve reinnervation of the dermal graft. In rats, this construct has demonstrated the generation of compound muscle action potentials (CMAPs), amplifying the target nerve&#39;s signal from the micro- to milli-volt level, with signal to noise ratios averaging approximately 30-50. Stimulation of the dermal component of the construct generates compound sensory nerve action potentials (CSNAPs) at the proximal nerve. As such, this construct has promising future utility towards the realization of the ideal, intuitive prosthetic.

Fabrication of the Composite Regenerative Peripheral Nerve Interface C-RPNI in the Adult Rat

The following manuscript describes a novel method for developing a biologic, closed loop neural feedback system termed the composite regenerative peripheral nerve interface (C-RPNI). This construct has the ability to integrate with peripheral nerves to amplify efferent motor signals while simultaneously providing afferent sensory feedback.

Herstellung der Composite Regenerative Peripheral Nerve Interface (C-RPNI) in der Erwachsenenratte

Recent advances in neuroprosthetics have enabled those living with extremity loss to reproduce many functions native to the absent extremity, and this is ...

Methods Article

20 mJ, 1 ps Yb: YAG Dünnschicht-Regenerationsverstärker