Gostaríamos de agradecer ao Prof. Ferenc Krausz pelas discussões e Najd Altwaijry por seu apoio à finalização do manuscrito. Este trabalho foi financiado pelo Centro de Aplicações Laser Avançadas (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>

Os autores não têm nada a divulgar.

A operação turn-key do oscilador é alcançada pelo ótimo gerenciamento de calor dos diferentes componentes do laser. A saída do oscilador é reproduzível diariamente, sem necessidade de alinhamento ou otimização extra. Além disso, a estabilidade de energia de impulso a pulso ea estabilidade espacial apontadora do laser de sementes cumprem as condições prévias para alcançar o funcionamento estável do amplificador regenerativo. Outras fontes de semente de baixa energia, como amplificadores de fibra, podem ser usadas para semear o amplificador. Neste estudo, utilizou-se um oscilador KLM de 2 μJ Yb: YAG fino para auxiliar a amplificação do amplificador regenerativo, reduzindo o crescimento das fases não-lineares acumuladas, uma vez que o número necessário de viagens de ida e volta é reduzido para a energia de sementes de maior entrada . Além disso, a maior energia de sementes influencia o processo de amplificação e reduz o estreitamento do ganho. A largura de banda espectral medida do impulso amplificadoEs para diferentes energias de sementes com uma potência de bomba fixa é mostrado na Figura 5c . A largura de banda espectral amplificada diminui para menores energias de sementes devido ao estreitamento do ganho. Para 10 pJ de energia de sementes, o laser opera no período de duplicação, e não é possível alcançar uma operação estável, mesmo aumentando o número de viagens de ida e volta. Além da otimização cuidadosa dos sistemas de refrigeração e da fonte de alimentação dos diodos, a operação do amplificador regenerativo em saturação desempenha um papel importante na estabilidade alcançada do amplificador. O fundamental ou o segundo harmônico do laser podem ser usados ​​para bombear um sistema OPCPA. Para SHG, as performances de um LBO e um BBO foram comparadas, pois oferecem alto coeficiente não linear e limiar de dano, apesar da maior distância espacial e da abertura disponível limitada no caso do BBO. Como o coeficiente não-linear de BBO é quase o dobro do LBO, um cristal mais curto é suficientePara atingir o limite de saturação para SHG ( Figura 6a ). Portanto, BBO é a escolha mais adequada, já que a fase não linear linear acumulada é menor 28 . As durações de pulso dos pulsos SH são caracterizadas experimentalmente em diferentes eficiências de conversão. Observou-se que, com altas eficiências de conversão, o espectro SHG é ampliado e uma fase espectral de ordem superior aparece ( Figura 6 ). Portanto, o caso B, com a eficiência de conversão de 70%, é escolhido onde o SH e os feixes fundamentais não convertidos mantêm excelente qualidade.

A geração de pulsos de laser de alta energia e poucos ciclos com alta taxa de repetição é de grande interesse para os campos aplicados, como attosecond science 1 , 2 , 3 , 4 e física de campo alto 5 , 6 , que podem se beneficiar diretamente Da disponibilidade de tais fontes. O OPCPA representa a rota mais promissora para alcançar energias de pulso elevadas e amplitudes de banda de amplificação amplas que suportam simultaneamente impulsos de poucos ciclos 1 . Até à data, OPCPA permite amplificação de banda larga ultra, o que gera impulsos de poucos ciclos 7 , 8 , 9 , 10 . No entanto, uma implementação modificada do esquema OPCPA, que usa pulsos de bomba curtos na escala de picoseconda, é promissora paraTornando esta abordagem escalável para energias de pulso ainda maiores e poderes médios no regime de poucos ciclos 1 , 11 , 12 . Devido à alta intensidade da bomba no OPCPA bombeado por pulso curto, o ganho de passagem única alta permite o uso de cristais muito finos para suportar grandes largura de banda de amplificação. Embora o OPCPA bombeado com pulso curto tenha muitas vantagens, a realização dessa abordagem está sujeita à disponibilidade de lasers especialmente adaptados para esse fim. Essas lâminas de bomba são necessárias para fornecer pulsos de picossegundo de alta energia com qualidade de feixe de quase difracção a taxas de repetição na faixa de kHz a MHz 13 , 14 , 15 . A introdução de lasers dopados com iterbério em geometrias diferentes, capazes de fornecer pulsos de laser de picosecond com alta energia e alta potência média, Estão prestes a mudar o estado atual do campo 1 , 13 , 14 , 15 , 16 , 17 , 18 . Yb: YAG tem boa condutividade térmica e uma longa vida útil do estado superior, e pode ser bombeada por laser de diodo com custo efetivo. Seu desempenho quando usado na geometria do disco fino é excelente devido ao resfriamento eficiente do meio de ganho para escalar simultaneamente o pico e a potência média. Além disso, a ocorrência de auto-focagem dentro do meio de ganho durante o processo de amplificação é suprimida devido à esbeltez do disco fino em comparação com outras geometrias de média de ganho, resultando em excelentes perfis temporais e espaciais dos pulsos amplificados. Combinar esse conceito com a CPA é promissor para gerar pulsos de picosegundo com centenas de milinholos de energia e centenasDe watts de potência média 19 , 20 . O objetivo deste trabalho é demonstrar um amplificador regenerativo de disco fino Yb: YAG com excelente performance diária como fonte adequada para bombeamento de OPCPAs 21 . Para atingir esse objetivo, este estudo emprega um oscilador de disco fino Yb: YAG 22 com vários microjogos de energia de pulso para semear o amplificador para minimizar a fase não linear linear acumulada durante o processo de amplificação. Este protocolo fornece a receita para a construção e operação do sistema a laser, que é descrito em outro lugar 21 . São apresentados detalhes sobre software de implementação e controle de componentes, e o processo de alinhamento do sistema é descrito.

<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

Este é um relatório sobre um amplificador regenerativo de 100 W, 20 mJ, 1 ps Yb: YAG de disco fino. Um microfone caseiro Yb: YAG, o oscilador bloqueado em modo Kerr-lens com desempenho turn-key e energia de pulso de nível microjolar é usado para semear o amplificador de pulso chirpado regenerativo. O amplificador é colocado em uma caixa hermética. Ele opera a temperatura ambiente e exibe operação estável a uma taxa de repetição de 5 kHz, com uma estabilidade de pulso a pulso inferior a 1%. Ao empregar um cristal de borato de bário beta de 1,5 mm de espessura, a freqüência da saída do laser é dobrada para 515 nm, com uma potência média de 70 W, o que corresponde a uma eficiência óptica a óptica de 70%. Esta performance superior torna o sistema uma fonte de bomba atrativa para amplificadores paramétricos ópticos de pulso-piloto na faixa espectral do infravermelho próximo e infravermelho médio. Combinando o desempenho turn-key e a estabilidade superior do amplificador regenerativo, o sistema facilita a geração de uma banda larga, CEP-stablesemente. Fornecer a semente e a bomba da amplificação paramétrica de pulsos paramétricos (OPCPA) de uma fonte laser elimina a demanda de sincronização temporal ativa entre esses pulsos. Este trabalho apresenta um guia detalhado para configurar e operar um amplificador regenerativo de disco fino Yb: YAG, baseado em amplificação de pulso chirpado (CPA), como fonte de bomba para um amplificador paramétrico de pulso polarizado.

<html> O oscilador entrega pulsos de 350 fs, 2 μJ, 25 W à taxa de repetição de 11 MHz, com uma estabilidade de pulso a pulso de 1% (rms) e flutuações apontadoras de feixe de menos de 0,6% ao longo de 1 h de medição ( Figura 4 ). <img alt="Figura 4" class="xfigimg" src="/files/ftp_upload/55717/55717fig4.jpg" /> Figura 4 : Yb: YAG de disco fino, oscilador bloqueado pelo modo Kerr-lens. (A) O espectro (vermelho), o perfil recuperado temporais intensidade (azul), e o perfil espacial (inset) dos impulsos do oscilador. ( B ) Espectrógrafo SHG-FROG medido e recuperado do oscilador. Esta figura foi modificada de Fattahi et al. , Com permissão da referência 21 .&gt; Clique aqui para ver uma versão maior desta figura. Os impulsos de semente são amplificados no amplificador regenerativo para 125 W enquanto são bombeados com um diodo acoplado de fibra CW a um comprimento de onda de 940 nm a 280 W, correspondendo a uma eficiência óptica-óptica de 47%. A estabilidade de pulso a pulso do amplificador é inferior a 1%, e o amplificador apresenta excelente estabilidade a longo prazo após 10 h de operação contínua. O feixe amplificado tem um excelente perfil espacial, com um M 2 de 1 (M 2 x = 1,08 e M 2 y = 1,07) e um excelente perfil temporal após compressão para 1 ps (em FWHM) ( Figura 5 ). <img alt="Figura 5" class="xfigimg" src="/files/ftp_upload/55717/55717fig5.jpg" /> Figura 5 : Caracterização do amplificador regenerativoSaída e o efeito de redução de ganho. (A) A estabilidade do amplificador de potência média regenerativo após 10 horas de operação contínua. Inserção: ( a-1 ) Potência normalizada para o seu valor médio em uma janela de tempo de 0,5 h; (A -2 ) Perfil de feixe de saída do amplificador regenerativo. ( B ) Espectro de saída do amplificador (verde) e a intensidade temporal recuperada (azul) dos impulsos do laser a uma potência média de 100 W após o compressor de rede. ( C ) Energia de sementes versus largura de banda espectral (FWHM) da saída do amplificador e as viagens de ida e volta necessárias para a mesma potência média de saída a 300 W da potência da bomba. Esta figura foi modificada de Fattahi et al. , Com permissão da referência 21 . <a href="http://ecsource.jove.com/files/ftp_upload/55717/55717fig5large.jpg" target="_blank">Clique aqui para ver uma versão maior dessa figura.</a> 25 . Foram considerados dois cristais diferentes com os seguintes parâmetros: 1) um triborato de lítio de tipo I, 6 mm de espessura (LBO), com um ângulo de correspondência de fase de 13,7 ° e um coeficiente não linear de 0,819 pm / V e 2) um BBO tipo I, de 3 mm de espessura com um ângulo de correspondência de fase de 23,4 ° e um coeficiente não linear de 2 pm / V 26 , 27 . 1-ps, pulsos de 20-mJ a 1.030 nm e uma intensidade de pico de 100 GW / cm 2 foram considerados como a entrada da simulação. Os resultados da simulação mostraram que o desempenho do BBO foi superior ao da LBO para SHG ( Figura 6 ). <img alt="Figura 6" class="xfigimg" src="/files/ftp_upload/55717/55717fig6.jpg" /> Figura 6 : Geração de segunda harmônica. (A) Simulado SHG enoRgy para um cristal LBO de 6 mm de espessura e um cristal BBO de 3 mm de espessura. ( B ) eficiência de SHG experimental versus intensidade de pico da bomba de entrada em um cristal BBO de 1,5 mm de espessura usando 0,5 mJ (preto) e 20 mJ (verde) da saída do amplificador. ( C ) A intensidade espectral recuperada e ( d ) o atraso grupal das medidas XFROG para diferentes eficiências SHG correspondentes aos pontos A, B e C em (b). Esta figura foi modificada de Fattahi et al. , Com permissão da referência 21 . <a href="http://ecsource.jove.com/files/ftp_upload/55717/55717fig6large.jpg" target="_blank">Clique aqui para ver uma versão maior dessa 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

Um protocolo para a operação de uma fonte de bomba de amplificador de pulso paramétrico óptico de alta potência, de alta potência, de alta potência com base em um amplificador regenerativo de disco fino Yb: YAG é apresentado aqui.

20 mJ, 1 ps Yb: amplificador regenerativo de disco fino YAG

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 visualizações.  Ludwig Maximilian University of Munich.  Um protocolo para a operação de uma fonte de bomba de amplificador de pulso paramétrico óptico de alta potência, de alta potência, de alta potência com base em um amplificador regenerativo de disco fino Yb: YAG é apresentado aqui. 

Vídeo: 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 ...

Bioengenharia

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 Nanopartículas Regenerativa Research

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.

Geração de espessura completa tridimensional Pele Equivalente 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.

Neonatal Cardiac Scaffolds: Matrizes novas para Estudos Regenerativa

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.

Estudar Soft-matéria e Sistemas Biológicos longo de um comprimento escala larga de nanômetros e micrômetro Tamanhos no pequeno ângulo Neutron Difractï¿½etro 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 para terapia regenerativa injetável e triagem de drogas de alto rendimento

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.

Protocolo genérico para otimização da produção de proteína heteróloga usando tecnologia automatizada de 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.

Recombinante colágeno peptídeo Microcarriers para expansão da célula e seu potencial uso como sistema de entrega de células em um bioreator modelo

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.

Construção de uma folha de células-tronco mesenquimais em várias camadas com um sistema de cultura dinâmica 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.

Fabricação da Interface Nervosa Periférica Regenerativa Composta (C-RPNI) no Rato Adulto

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