Vorremmo ringraziare il prof. Ferenc Krausz per le discussioni e Najd Altwaijry per il suo supporto per la finalizzazione del manoscritto. Questo lavoro è stato finanziato dal Centro per le Applicazioni Laser avanzate (CALA).

<ol>
	<li>Fattahi, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Third-generation+femtosecond+technology.">Third-generation femtosecond technology.</a> Optica. 1 (1), 45-63 (2014).</li><li>Hentschel, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Attosecond+metrology.">Attosecond metrology.</a> Nature. 414 (6863), 509-513 (2001).</li><li>Cavalieri, A. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intense+1.5-cycle+near+infrared+laser+waveforms+and+their+use+for+the+generation+of+ultra-broadband+soft-x-ray+harmonic+continua.">Intense 1.5-cycle near infrared laser waveforms and their use for the generation of ultra-broadband soft-x-ray harmonic continua.</a> New J Phys. 9 (7), 242 (2007).</li><li>Schweinberger, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Waveform-controlled+near-single-cycle+milli-joule+laser+pulses+generate+sub-10+nm+extreme+ultraviolet+continua.">Waveform-controlled near-single-cycle milli-joule laser pulses generate sub-10 nm extreme ultraviolet continua.</a> Opt Lett. 37, 3573-3575 (2012).</li><li>Buck, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Real-time+observation+of+laser-driven+electron+acceleration.">Real-time observation of laser-driven electron acceleration.</a> Nature Phys. 7 (7), 543-548 (2011).</li><li>Zhong, H., Karpowicz, N., Zhang, X. 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. X., Kong, Y. F., Zhang, J. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Optical+parametric+properties+of+532-nm-pumped+beta-barium-borate+near+the+infrared+absorption+edge.">Optical parametric properties of 532-nm-pumped beta-barium-borate near the infrared absorption edge.</a> Opt Commun. 184 (5), 485-491 (2000).</li><li>Kato, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Temperature-tuned+90o+phase-matching+properties+of+LiB3O5.">Temperature-tuned 90o phase-matching properties of LiB3O5.</a> IEEE J Quant Electron. 30 (12), 2950-2952 (1994).</li><li>Fattahi, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Third-generation femtosecond technology. , (2016).</li></ol>

Gli autori non hanno nulla da rivelare.

Il funzionamento a chiave del oscillatore è ottenuto dalla gestione ottimale del calore delle diverse componenti del laser. L&#39;uscita dell&#39;oscillatore è riproducibile su base giornaliera, senza necessità di allineamento o ottimizzazione supplementari. Inoltre, la stabilità energetica impulso-impulso e la stabilità di punta spaziale del laser di sementi soddisfano le condizioni preliminari per il funzionamento stabile dell&#39;amplificatore rigenerativo. Altre sorgenti di sementi a basso contenuto energetico, come gli amplificatori a fibre, possono essere utilizzati per seminare l&#39;amplificatore. In questo studio è stato utilizzato un oscillatore KLM da 2 μJ Yb: YAG per aiutare l&#39;amplificazione dell&#39;amplificatore rigenerativo riducendo la crescita delle fasi non lineari accumulate, in quanto il numero di giri voluto richiesto è ridotto per l&#39;energia di semi più alta . Inoltre, la maggiore energia seme influenza il processo di amplificazione e riduce il rallentamento del guadagno. La larghezza di banda spettrale misurata del pulso amplificatoEs per energie di sementi differenti a una potenza fissa della pompa è mostrato in Figura 5c . La larghezza di banda spettrale amplificata diminuisce per le energie di seme inferiori a causa del restringimento dei guadagni. Per l&#39;energia di sementi da 10 pJ, il laser opera nel periodo raddoppiato e non è possibile raggiungere un funzionamento stabile, anche aumentando il numero di giri andati. Oltre all&#39;attenta ottimizzazione dei sistemi di raffreddamento e dell&#39;alimentazione dei diodi, il funzionamento dell&#39;amplificatore rigenerativo a saturazione svolge un ruolo importante nella stabilità raggiunta dell&#39;amplificatore. La fondamentale o la seconda armonica del laser può essere usata per pompare un sistema OPCPA. Per SHG, sono state confrontate le prestazioni di un LBO e un cristallo di BBO, in quanto offrono un elevato coefficiente non lineare e una soglia di danno, nonostante il largo spazio spaziale e l&#39;apertura limitata disponibile nel caso della BBO. Poiché il coefficiente non lineare della BBO è quasi il doppio del LBO, un cristallo più corto è sufFinient per raggiungere il limite di saturazione per SHG ( Figura 6a ). Pertanto, BBO è la scelta più adatta, poiché la fase non lineare accumulata è più piccola 28 . Le durate di impulso degli impulsi SH sono caratterizzate sperimentalmente a diverse efficienze di conversione. È stato osservato che ad alta efficienza di conversione, lo spettro SHG è ampliato e appare una fase spettrale di ordine superiore ( Figura 6 ). Pertanto viene scelto il caso B, con l&#39;efficienza di conversione del 70%, dove gli SH e le travi fondamentali non convertite mantengono una qualità eccellente.

La generazione di impulsi laser ad alta energia, a pochi cicli ad elevata frequenza di ripetizione, è di grande interesse per i campi applicati, come la scienza attosecond 1 , 2 , 3 , 4 e la fisica alta 5 , 6 , Dalla disponibilità di tali fonti. OPCPA rappresenta la via più promettente per ottenere energie ad impulso elevate e ampie larghezze di banda di amplificazione che supportano contemporaneamente impulsi a pochi cicli 1 . Ad oggi, OPCPA consente l&#39;amplificazione ultra-banda larga, che genera impulsi a pochi cicli 7 , 8 , 9 , 10 . Tuttavia, un&#39;implementazione modificata dello schema OPCPA, che utilizza impulsi corti della pompa sulla scala di picosecondi, è promessa perRendendo questo approccio scalabile per energie di impulso ancora più elevate e potenze medie nel regime a pochi cicli 1 , 11 , 12 . A causa dell&#39;elevata intensità della pompa in OPCPA pompato a breve impulso, l&#39;elevato guadagno a singolo passaggio consente l&#39;utilizzo di cristalli molto sottili per supportare ampie larghezze di banda di amplificazione. Sebbene l&#39;OPCPA pompato a impulsi corti abbia molti vantaggi, la realizzabilità di questo approccio è soggetta alla disponibilità di laser adatti a questo scopo. Questi laser di pompaggio sono necessari per fornire impulsi picosecondi ad alta energia con qualità del fascio limitato vicino a diffrazione a velocità di ripetizione nella gamma da kHz a MHz 13 , 14 , 15 . L&#39;introduzione di laser a base di itterbium a diverse geometrie, in grado di fornire impulsi laser a picosecondi ad alta energia e ad alta potenza media, Stanno per cambiare lo stato attuale del campo 1 , 13 , 14 , 15 , 16 , 17 , 18 . Yb: YAG ha una buona conducibilità termica e una lunga durata di stato superiore, e può essere pompata da laser a diodo a basso costo. La sua prestazione quando viene utilizzata in geometria a disco sottile è eccezionale a causa del raffreddamento efficiente del mezzo di guadagno per scalare simultaneamente il picco e la potenza media. Inoltre, il fenomeno dell&#39;autofocusione all&#39;interno del mezzo di guadagno durante il processo di amplificazione viene soppresso a causa della maneggevolezza del disco sottile rispetto ad altre geometrie del medium di guadagno, con conseguente profili temporali e spaziali eccellenti degli impulsi amplificati. Combinando questo concetto con CPA promette di generare impulsi picosecondi con centinaia di million di energia e centinaiaDi watt di potenza media 19 , 20 . Lo scopo di questo lavoro è quello di dimostrare un amplificatore rigenerativo a disco rigido Yb: YAG con eccezionali prestazioni quotidiane come una fonte adatta per il pompaggio di OPCPA 21 . Per raggiungere questo obiettivo, questo studio utilizza un oscillatore 22 a disco Yb: YAG con microspi di energia a impulsi per seminare l&#39;amplificatore per minimizzare la fase non lineare accumulata durante il processo di amplificazione. Questo protocollo fornisce la ricetta per la costruzione e il funzionamento del sistema laser, descritto altrove 21 . Sono mostrati dettagli sul software di implementazione e controllo dei componenti e viene descritto il processo di allineamento del sistema.

<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

Si tratta di un rapporto su un amplificatore rigenerativo a disco sottile da 100 W, 20 mJ, 1 ps Yb: YAG. Un sottile disco Yb: YAG in forma casalinga, l&#39;oscillatore bloccato in modalità Kerr a obiettivo con prestazioni a chiave e energia a impulsi a micro-jusle viene utilizzato per seminare l&#39;amplificatore rigenerativo a impulsi a chirped. L&#39;amplificatore è collocato in un alloggiamento ermetico. Funziona a temperatura ambiente e presenta un funzionamento stabile a una frequenza di ripetizione di 5 kHz, con una stabilità impulso-impulso inferiore all&#39;1%. Utilizzando un cristallo di borato di beta borato di spessore di 1,5 mm, la frequenza dell&#39;uscita laser è raddoppiata a 515 nm, con una potenza media di 70 W, che corrisponde ad un&#39;efficienza ottica-ottica del 70%. Questa prestazione superiore rende il sistema una sorgente attraente della pompa per gli amplificatori parametrici a impulsi a impulsi ottici nell&#39;intervallo spettrale a infrarossi ea medio infrarossi. Combinando le prestazioni del turn-key e la stabilità superiore dell&#39;amplificatore rigenerativo, il sistema facilita la generazione di una banda larga, stabile CEPseme. Fornire la sorgente e la pompa dell&#39;amplificazione ottica parametrica a impulsi a chirped (OPCPA) da una sorgente laser elimina la richiesta di sincronizzazione temporale attiva tra questi impulsi. Questo lavoro presenta una guida dettagliata per l&#39;impostazione e l&#39;azionamento di un amplificatore rigenerativo a disco sottile Yb: YAG, basato su amplificazione a impulsi a chirpaggio (CPA), come sorgente della pompa per un amplificatore ottico a impulsi parametrici ottici.

<html> L&#39;oscillatore fornisce 350 fs, 2 μJ, impulsi a 25 W con frequenza di ripetizione di 11 MHz, con una stabilità impulso di impulso pari a 1% (rms) e fluttuazioni a raggio inferiore di 0,6% per 1 h di misura ( Figura 4 ). <img alt="Figura 4" class="xfigimg" src="/files/ftp_upload/55717/55717fig4.jpg" /> Figura 4 : Yb: disco sottile YAG, oscillatore bloccato in modalità Kerr-lens. (A) Lo spettro (rosso), il profilo temporale recuperato intensità (blu), e il profilo spaziale (riquadro) degli impulsi dell&#39;oscillatore. ( B ) misurato e recuperato lo spettrografo SHG-FROG dell&#39;oscillatore. Questa cifra è stata modificata da Fattahi et al. , Con permesso di riferimento 21 .&gt; Clicca qui per visualizzare una versione più grande di questa figura. Gli impulsi di semi vengono amplificati nell&#39;amplificatore rigenerativo fino a 125 W mentre vengono pompati con un diodo a fibre ottiche CW a una lunghezza d&#39;onda di 940 nm a 280 W, corrispondente ad un ottico-ottico di efficienza del 47%. La stabilità impulso-impulso dell&#39;amplificatore è inferiore all&#39;1% e l&#39;amplificatore presenta una stabilità a lungo termine eccellente dopo 10 ore di funzionamento continuo. La trave amplificato ha un eccellente profilo spaziale, con M 2 di 1 (M 2 x = 1,08 e M 2 y = 1,07) e un eccellente profilo temporale dopo la compressione a 1 ps (a FWHM) (Figura 5). <img alt="Figura 5" class="xfigimg" src="/files/ftp_upload/55717/55717fig5.jpg" /> Figura 5 : Caratterizzazione dell&#39;amplificatore rigenerativoUscita e l&#39;effetto di restringimento del guadagno. (A) La stabilità dell&#39;amplificatore rigenerativo potenza media dopo 10 ore di funzionamento continuo. Inset: ( a-1 ) Potenza normalizzata al suo valore medio in una finestra temporale di 0,5 h; (A -2 ) Profilo fascio di uscita dell&#39;amplificatore rigenerativo. ( B ) Spettro di uscita dell&#39;amplificatore (verde) e l&#39;intensità temporale recuperata (blu) dell&#39;impulso del laser a 100 W di potenza media dopo il compressore della griglia. ( C ) Energia di semi rispetto alla larghezza di banda spettrale (FWHM) dell&#39;uscita dell&#39;amplificatore e dei giri richiesti per la stessa potenza media di uscita a 300 W di potenza della pompa. Questa cifra è stata modificata da Fattahi et al. , Con permesso di riferimento 21 . <a href="http://ecsource.jove.com/files/ftp_upload/55717/55717fig5large.jpg" target="_blank">Clicca qui per visualizzare una versione più grande di questa figura.</a> 25 . Sono stati considerati due diversi cristalli con i seguenti parametri: 1) un triborato di litio (LBO) di tipo I, 6 mm, con un angolo di corrispondenza di fase di 13,7 ° e un coefficiente non lineare di 0,819 pm / V; e 2) Tipo I, BBO di 3 mm di spessore con un angolo di corrispondenza di fase di 23,4 ° e un coefficiente non lineare di 2 pm / V 26 , 27 . 1-ps, impulsi di 20 mJ a 1.030 nm e un&#39;intensità di picco di 100 GW / cm 2 sono stati considerati come l&#39;ingresso della simulazione. I risultati della simulazione hanno mostrato che la performance BBO era superiore a quella del LBO per SHG ( Figura 6 ). <img alt="Figura 6" class="xfigimg" src="/files/ftp_upload/55717/55717fig6.jpg" /> Figura 6 : Seconda generazione armonica. (A) simulato SHG enePer un cristallo LBO di 6 mm e un cristallo BBO di 3 mm di spessore. B ) Efficienza sperimentale SHG rispetto all&#39;intensità di picco della pompa di ingresso in un cristallo BBO di 1,5 mm di spessore usando 0,5 mJ (nero) e 20 mJ (verde) dell&#39;uscita dell&#39;amplificatore. ( C ) l&#39;intensità spettrale recuperata e ( d ) il ritardo del gruppo delle misurazioni XFROG per diverse efficienze SHG corrispondenti ai punti A, B e C in (b). Questa cifra è stata modificata da Fattahi et al. , Con permesso di riferimento 21 . <a href="http://ecsource.jove.com/files/ftp_upload/55717/55717fig6large.jpg" target="_blank">Clicca qui per visualizzare una versione più grande di questa figura.</a>

Watch this Scientific Journal Video about 20 mJ, 1 ps Yb:YAG Thin-disk Regenerative Amplifier at JoVE.com

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

Viene presentato un protocollo per l&#39;azionamento di una sorgente ottica ad alta potenza e ad alta potenza della pompa ad amplificatore a impulsi a impulsi parametrici basati su un amplificatore rigenerativo a disco rigido Yb: YAG.

20 mJ, 1 ps Yb: YAG Amplificatore rigenerativo a disco rigido

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 Views.  Ludwig Maximilian University of Munich.  Viene presentato un protocollo per l'azionamento di una sorgente ottica ad alta potenza e ad alta potenza della pompa ad amplificatore a impulsi a impulsi parametrici basati su un amplificatore rigenerativo a disco rigido Yb: YAG. 

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

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 Nanoparticelle for Regenerative ricerca

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.

Generazione di una tridimensionale a tutto spessore della pelle Equivalent e Automated Ferimento

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.

Neonatale cardiaco Impalcature: Matrici innovativi per gli Studi rigenerativi

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.

Studiare Soft-materia e sistemi biologici per una lunghezza scala larga da nanometri e micrometro Taglie al piccolo angolo Neutron Diffrattometro 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.

Microtissues 3D per la terapia rigenerativa iniettabile e droga di High-throughput 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.

Protocollo generico per l'ottimizzazione della produzione di proteine eterologhe utilizzando tecnologia automatizzata di Microbioreactor

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.

Ricombinante collagene Peptide Microcarriers per l'espansione delle cellule e loro potenziale utilizzo come sistema di consegna di cella in un bioreattore modello

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.

Costruzione di un foglio di multistrato delle cellule staminali mesenchimali con un sistema di cultura dinamica 3D

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.

Fabbricazione dell'interfaccia nervo periferica rigenerativa composita (C-RPNI) nel ratto adulto

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