ברצוננו להודות לפרופ &#39;פרנץ קראוס על הדיונים ועל נאג&#39;ד אלטוואיג&#39;רי על תמיכתה בסיום כתב היד. עבודה זו מומנה על ידי המרכז ליישומי לייזר מתקדמים (CALA).

<ol>
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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>

למחברים אין מה לגלות.

פעולת Turn-Key של המתנד מושגת על ידי ניהול החום האופטימלי של הרכיבים השונים של הלייזר. הפלט של מתנד הוא לשחזור על בסיס יומי, ללא צורך יישור נוסף או אופטימיזציה. בנוסף, יציבות הדופק אל הדופק ויציבות הצבעה מרחבית של לייזר הזרע ממלאת את התנאים המוקדמים להשגת הפעולה היציבה של מגבר התחדשות. מקורות אחרים של אנרגיית אנרגיה נמוכה, כגון מגברי סיבים, יכולים לשמש לזרז את המגבר. במחקר זה, מתנד KLM K2M בעל 2 מיקרוגרם YB: YAG נעשה שימוש כדי להגביר את ההגברה של המגבר המשובי על ידי הקטנת הצמיחה של השלבים הלא ליניאריים המצטברים, שכן המספר הדרוש של הנסיעות הלוך ושוב מופחת לאנרגיה גבוהה יותר של זרעי קלט . בנוסף, האנרגיה זרע גבוהה משפיע על תהליך ההגברה ומקטין את הצטמקות רווח. רוחב הפס הספקטראלי הנמדד של הפולסים המוגבריםEs עבור אנרגיות זרע שונות על כוח משאבה קבוע מוצג באיור 5 ג . רוחב פס מוגדל של רוחב פס מופחת עבור אנרגיות זרע נמוכות יותר בגלל צמצום הצטברות. עבור 10 pJ אנרגיית זרע, לייזר פועלת בתקופה הכפלה, ולא ניתן להגיע לפעולה יציבה, גם על ידי הגדלת מספר הסיורים הלוך ושוב. בנוסף לאופטימיזציה זהירה של מערכות הקירור ואת אספקת החשמל של דיודות, הפעולה של מגבר משובי רוויה משחק תפקיד מרכזי ביציבות שהושגה של המגבר. הרמוני הבסיסי או השני של הלייזר יכול לשמש משאבת מערכת OPCPA. עבור SHG, ההשוואות של LBO ו קריסטל BBO הושוו, כפי שהם מציעים מקדם לא לינארית גבוהה סף נזק, למרות הליכה מרחבית גדול יותר הצמצם זמין מוגבל במקרה של BBO. כמו מקדם הלא לינארית של BBO הוא כמעט כפול מזה של LBO, קריסטל קצר יותר הוא sufמתוחכם להגיע לגבול הרוויה עבור SHG ( איור 6 א ). לכן, BBO הוא הבחירה המתאימה יותר, כמו השלב הלא ליניארי שנצבר הוא קטן יותר 28 . משכי הדופק של הפולסים SH מאופיינים בניסוי ביעילות המרה שונה. זה נצפתה כי יעילות המרה גבוהה, הספקטרום SHG הוא הרחיב ואת שלב גבוה ספקטרלי מופיע ( איור 6 ). לכן, במקרה B, עם יעילות ההמרה של 70%, נבחר שבו SH ואת הקורות היסוד לא משמרת לשמור על איכות מעולה.

הדור של אנרגיה גבוהה, כמה מחזורי פולסים לייזר בקצב גבוה החזרה הוא עניין רב תחומים מיושמים, כגון מדע אטוסקונד 1 , 2 , 3 , 4 ו פיזיקה שדה גבוה 5 , 6 , אשר עומדים ליהנות ישירות מן הזמינות של מקורות כאלה. OPCPA מייצג את התוואי המבטיח ביותר להשגת אנרגיות דופק גבוהות ורוחבי הגברה גדולים, אשר תומכים בו זמנית במספר פעימות מחזוריות 1 . עד כה, OPCPA מאפשר הגברה אולטרה פס רחב, אשר מייצר כמה מחזורי פעימות 7 , 8 , 9 , 10 . עם זאת, יישום שונה של ערכת OPCPA, אשר משתמש פולסים משאבה קצרה על סולם picosond, מחזיקה הבטחה עבורמה שהופך גישה זו להרחבה עבור אנרגיות הדופק אפילו גבוה יותר ואת הכוחות הממוצעים במעגל כמה מחזור 1 , 11 , 12 . בשל עוצמת המשאבה הגבוהה בדופק קצר OPCPA, העלייה החד-פעמית הגבוהה מאפשרת שימוש בגבישים דקים מאוד לתמיכה ברוחבי הגברה גדולים. למרות הדופק קצר שאוב OPCPA יש יתרונות רבים, realisability של גישה זו כפופה לזמינות של לייזרים המותאמים במיוחד למטרה זו. אלה לייזרים משאבה נדרשים לספק אנרגיה גבוהה picosond פולסים עם כמעט עקיפה איכות קרן מוגבל בשיעורי החזרה ב kHz ל MHz טווח 13 , 14 , 15 . ההקדמה של לייזרים ytterbium מסוממים על גיאומטריות שונות, מסוגל לספק פולסים לייזר picosond עם אנרגיה גבוהה כוח ממוצע גבוה, עומדים לשנות את המצב הנוכחי של השדה 1 , 13 , 14 , 15 , 16 , 17 , 18 . Yb: YAG יש מוליכות תרמית טובה חיים ארוכים המדינה העליונה, וזה יכול להיות שאוב על ידי לייזרים דיודה חסכונית. הביצועים שלה כאשר נעשה שימוש בגיאומטריה דק הדיסק הוא יוצא מן הכלל בשל קירור יעיל של המדיום רווח כדי בו זמנית בקנה מידה את השיא ואת הכוח הממוצע. יתר על כן, המופע של התמקדות עצמית בתוך המדיום רווח במהלך תהליך ההגברה הוא מדוכא בשל הדקיקות של הדיסק דק בהשוואה גיאומטריות אחרות בינונית רווח, וכתוצאה מכך פרופילים הזמני והמרחבי מעולה של הפולסים מוגבר. שילוב המושג הזה עם רו&quot;ח מקיים הבטחה ליצירת פעימות פיקו-שנייה עם מאות מילי-וולטים של אנרגיה ומאותשל וואט של הספק ממוצע 19 , 20 . מטרת עבודה זו היא להדגים מתג Yb: YAG רזה מגבר משובי דיסק עם ביצועים יומית יוצאת דופן כמקור מתאים שאיבה OPCPAs 21 . כדי להשיג מטרה זו, מחקר זה מעסיק Yb: YAG דיסק דק מתנד 22 עם כמה microjoules של אנרגיה הדופק זרע המגבר כדי למזער את השלב הלא ליניארי שנצבר במהלך תהליך ההגברה. פרוטוקול זה מספק את המתכון לבניית ותפעול מערכת לייזר, אשר מתואר במקום אחר 21 . פרטים על יישום רכיבים ותוכנה מלאה מוצגים, ותהליך היישור של המערכת מתואר.

<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

זהו דו&quot;ח על 100 W, 20 mJ, 1 PS Yb: YAG דיסק דק מגבר משובי. תוצרת בית Yb: YAG דיסק דק, מתנד נעול במצב Kerr- נעול עם ביצועים Turn-Key ו microjoule ברמת הדופק אנרגיה משמש הזרע מגבר צ &#39;יפס הדופק משובי. המגבר ממוקם בדיור אטום. הוא פועל בטמפרטורת החדר ומציג פעילות יציבה בקצב של 5 קילו-הרץ, עם יציבות הדופק לדופק פחות מ -1%. על ידי העסקת 1.5 מ&quot;מ עובי בטא בריום borate קריסטל, התדר של הפלט לייזר מוכפל ל 515 ננומטר, עם כוח ממוצע של 70 וואט, אשר תואם אופטי אופטי אופטי של 70%. ביצועים מעולים אלה הופכים את המערכת למקור משאבת אטרקטיבי עבור מגברים אופטיים פרמטריים צולבים-דופק בטווח ספקטרלי הקרוב אינפרא אדום ו באמצע אינפרא אדום. השילוב של ביצועי Turn-Key ויציבות מעולה של מגבר התחדשות, המערכת מאפשרת את יצירתו של פס רחב, CEP יציבזֶרַע. אספקת הזרע והמשאבה של הגברה אופטית של הדופק המוטבעי (OPCPA) ממקור לייזר אחד מבטלת את הדרישה לסינכרון הזמני הפעיל בין הפולסים האלה. עבודה זו מציגה מדריך מפורט להקמת ולהפעיל מגבר Yb: YAG דיסק רנרטיטיבי מחדש, המבוססת על הגברה הדופק הצפצופים (CPA), כמקור המשאבה של מגבר אופקי פרמטרי צ &#39;יפס הדופק.

<html> מתנד מספק 350 פולסים, 2 μJ, 25-W פולסים בקצב חזרות 11 MHz, עם יציבות הדופק לדופק של 1% (rms) ותנודות הצבעה קרן של פחות מ -0.6% מעל 1 שעות של מדידה ( איור 4 ). <img alt="איור 4" class="xfigimg" src="/files/ftp_upload/55717/55717fig4.jpg" /> איור 4 : Yb: YAG דיסק דק, Kerr- העדשה מצב נעול נעול. ( א ) הספקטרום (אדום), פרופיל העוצמה הטמפורית המאוחזרת (כחול) והפרופיל המרחבי (הבלעדי) של פעימות המתנד. ( ב ) נמדד ואחזר ספקטרוגרף SHG-FROG של מתנד. נתון זה שונה מ Fattahi et al. , באישור 21 .&gt; אנא לחץ כאן כדי להציג גרסה גדולה יותר של נתון זה. זרעי הזרע מוגברים במגבר משובי ל 125 W בזמן היותו נשאב עם דיודה CW מצמידים דיודה באורך גל של 940 ננומטר ב 280 W, המתאים אופטי אופטי אופטי של 47%. יציבות הדופק אל הדופק של המגבר נמוכה מ -1%, והמגבר מציג יציבות מצוינת לטווח הארוך לאחר 10 שעות של פעולה מתמשכת. הקורה המוגברת בעלת פרופיל מרחבי מעולה, עם M 2 של 1 (M 2 x = 1.08 ו- M 2 y = 1.07) ופרופיל טמפורלי מצוין לאחר דחיסה ל -1 ps (ב- FWHM) ( איור 5 ). <img alt="איור 5" class="xfigimg" src="/files/ftp_upload/55717/55717fig5.jpg" /> איור 5 : אפיון מגבר התחדשות<html> הפלט והאפקט המצמצם. ( א ) יציבות של מגבר התחדשות הממוצע כוח לאחר 10 שעות של פעולה מתמשכת. Inset: ( a-1 ) מנורמל כוח לערכו הממוצע בחלון זמן של 0.5 h; ( A-2 ) פלט פרופיל הקלט של מגבר משובי. ( ב ) ספקטרום פלט מגבר (ירוק) ואת העוצמה הזמנית המאוחזרת (כחול) של פולסים לייזר ב 100 W כוח הממוצע לאחר מדחס הסורג. ( ג ) אנרגיית זרע לעומת רוחב פס ספקטרלי (FWHM) של תפוקת המגבר והנסיעות העגולות הנדרשות עבור אותו הספק ממוצע של הספק ב -300 ואט של כוח משאבה. נתון זה שונה מ Fattahi et al. , באישור 21 . <a href="http://ecsource.jove.com/files/ftp_upload/55717/55717fig5large.jpg" target="_blank">אנא לחץ כאן כדי להציג גרסה גדולה יותר של דמות זו.</a> Hin-page = &quot;1&quot;&gt; ה- SHG נותח באמצעות קוד SISYFOS 25 . שני גבישים שונים עם הפרמטרים הבאים נחשבו: 1) שבב ליתיום מסוג I, 6 מ&quot;מ (LBO), עם זווית התאמת פאזה של 13.7 ° ומקדם לא ליניארי של 0.819 pm / V, ו- 2) סוג I, 3 מ&quot;מ עובי BBO עם זווית שלב ההתאמה של 23.4 ° ו מקדם לא לינארית של 2 pm / V 26 , 27 . 1-PS, 20-mJ פעימות ב 1,030 ננומטר ועוצמת שיא של 100 GW / cm 2 נחשבו קלט של הסימולציה. תוצאות הסימולציה הראו כי ביצועי ה- BBO היו טובים יותר מזו של ה- LBO עבור SHG ( איור 6 ). <img alt="איור 6" class="xfigimg" src="/files/ftp_upload/55717/55717fig6.jpg" /> איור 6 : הדור השני של ההרמוניה. ( א ) סימולציה SHG ​​מדומהRgy עבור גביש 6 מ&quot;מ עובי LBO ו 3 מ&quot;מ עובי BBO קריסטל. ( ב ) יעילות SHG ניסיוני לעומת עוצמת שיא שיא המשאבת ב 1.5 מ&quot;מ עובי BBO קריסטל באמצעות 0.5 mJ (שחור) ו 20 mJ (ירוק) של מגבר פלט. ( ג ) עוצמת ספקטרלית מאוחזרת ו ( ד ) עיכוב הקבוצה של מדידות XFROG עבור יעילות SHG שונים המתאימים לנקודות A, B, ו- C ב (ב). נתון זה שונה מ Fattahi et al. , באישור 21 . <a href="http://ecsource.jove.com/files/ftp_upload/55717/55717fig6large.jpg" target="_blank">אנא לחץ כאן כדי להציג גרסה גדולה יותר של דמות זו.</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

פרוטוקול להפעלה של אנרגיה גבוהה, כוח גבוה אופטית פרמטרית צייץ הדופק מגבר מקור משאבת מבוסס על Yb: YAG דיסק דק מגבר משובי מוצג כאן.

20 mJ, 1 PS Yb: YAG דיסק דק מגבר משובי

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.  פרוטוקול להפעלה של אנרגיה גבוהה, כוח גבוה אופטית פרמטרית צייץ הדופק מגבר מקור משאבת מבוסס על Yb: YAG דיסק דק מגבר משובי מוצג כאן. 

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

Elastomeric PGS Scaffolds in Arterial Tissue Engineering

Cardiovascular disease is one of the leading cause of mortality in the US and especially, coronary artery disease increases with an aging population and increasing obesity1. Currently, bypass surgery using autologous vessels, allografts, and synthetic grafts are known as a commonly used for arterial substitutes2. However, these grafts have limited applications when an inner diameter of arteries is less than 6 mm due to low availability, thrombotic complications, compliance mismatch, and late intimal hyperplasia3,4. To overcome these limitations, tissue engineering has been successfully applied as a promising alternative to develop small-diameter arterial constructs that are nonthrombogenic, robust, and compliant. Several previous studies have developed small-diameter arterial constructs with tri-lamellar structure, excellent mechanical properties and burst pressure comparable to native arteries5,6. While high tensile strength and burst pressure by increasing collagen production from a rigid material or cell sheet scaffold, these constructs still had low elastin production and compliance, which is a major problem to cause graft failure after implantation. Considering these issues, we hypothesized that an elastometric biomaterial combined with mechanical conditioning would provide elasticity and conduct mechanical signals more efficiently to vascular cells, which increase extracellular matrix production and support cellular orientation.
The objective of this report is to introduce a fabrication technique of porous tubular scaffolds and a dynamic mechanical conditioning for applying them to arterial tissue engineering. We used a biodegradable elastomer, poly (glycerol sebacate) (PGS)7 for fabricating porous tubular scaffolds from the salt fusion method. Adult primary baboon smooth muscle cells (SMCs) were seeded on the lumen of scaffolds, which cultured in our designed pulsatile flow bioreactor for 3 weeks. PGS scaffolds had consistent thickness and randomly distributed macro- and micro-pores. Mechanical conditioning from pulsatile flow bioreactor supported SMC orientation and enhanced ECM production in scaffolds. These results suggest that elastomeric scaffolds and mechanical conditioning of bioreactor culture may be a promising method for arterial tissue engineering.

Elastomeric PGS scaffolds with vascular smooth muscle cells cultured in a pulsatile flow bioreactor may lead to promising small-diameter arterial constructs with native ECM production in a relatively short culture period.

PGS אלסטומרי פיגומים להנדסת רקמות העורקים

Cardiovascular disease is one of the leading cause of mortality in the US and especially, coronary artery disease increases with an aging population and ...

Constructing a Collagen Hydrogel for the Delivery of Stem Cell-loaded Chitosan Microspheres

Multipotent stem cells have been shown to be extremely useful in the field of regenerative medicine1-3. However, in order to use these cells effectively for tissue regeneration, a number of variables must be taken into account. These variables include: the total volume and surface area of the implantation site, the mechanical properties of the tissue and the tissue microenvironment, which includes the amount of vascularization and the components of the extracellular matrix. Therefore, the materials being used to deliver these cells must be biocompatible with a defined chemical composition while maintaining a mechanical strength that mimics the host tissue. These materials must also be permeable to oxygen and nutrients to provide a favorable microenvironment for cells to attach and proliferate. Chitosan, a cationic polysaccharide with excellent biocompatibility, can be easily chemically modified and has a high affinity to bind with in vivo macromolecules4-5. Chitosan mimics the glycosaminoglycan portion of the extracellular matrix, enabling it to function as a substrate for cell adhesion, migration and proliferation. In this study we utilize chitosan in the form of microspheres to deliver adipose-derived stem cells (ASC) into a collagen based three-dimensional scaffold6. An ideal cell-to-microsphere ratio was determined with respect to incubation time and cell density to achieve maximum number of cells that could be loaded. Once ASC are seeded onto the chitosan microspheres (CSM), they are embedded in a collagen scaffold and can be maintained in culture for extended periods. In summary, this study provides a method to precisely deliver stem cells within a three dimensional biomaterial scaffold.

A major hurdle in current stem cell therapies is determining the most effective method to deliver these cells to host tissues. Here, we describe a chitosan-based delivery method that is efficient and simple in approach, while allowing adipose-derived stem cells to maintain their multipotency.

בניית Hydrogel קולגן עבור משלוח של התא טעון גזע Chitosan microspheres

Multipotent stem cells have been shown to be extremely useful in the field of regenerative medicine1-3. However, in order to use these cells effectively ...

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 ...

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.

הרמוני חלקיקים למחקר הרגנרציה

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.

דור של עור בעובי מלא תלת ממדי Equivalent ואוטומטי פציעה

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 לב פיגומים: מטריצות Novel ללימודי הרגנרציה

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.

לימוד-משנה רך מערכות ביולוגיות על פני אורך בקנה מידה רחבה מן ננומטר ו מיקרומטר גדלים בזווית-הקטנה ניוטרון 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. ...

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 תלת-ממד עבור טיפול בזריקות רגנרטיבית ותפוקה גבוהה והתרופות הקרנה

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.

פרוטוקול גנרי עבור אופטימיזציה של ייצור החלבון Heterologous באמצעות טכנולוגיית Microbioreactor אוטומטיות

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.