Name: low-cost custom fabrication and mode-locked operation of an all-normal-dispersion femtosecond fiber laser for multiphoton microscopy
Uploaded: 2019-11-22T12:30:00
Description: Watch this Scientific Journal Video about Low-cost Custom Fabrication and Mode-locked Operation of an All-normal-dispersion Femtosecond Fiber Laser for Multiphoton Microscopy at JoVE.com

We thank Drs. E. Cronin-Furman and M. Weitzman (Olympus Corporation of the Americas Scientific Solutions Group) for assistance in acquiring images. This work was supported by National Institutes of Health Grant K22CA181611 (to B.Q.S.) and the Richard and Susan Smith Family Foundation (Newton, M.A.) Smith Family Award for Excellence in Biomedical Research (to B.Q.S.).

1. Splice single mode fibers (SMF)
NOTE: Section 1 consists of general steps to splice SMFs. This is a non-essential, but recommended, step for practicing fiber splices using inexpensive fiber. This step ensures proper performance of the splicing equipment before using more valuable fiber optic materials.
<ol>
	<li>Cleave the first fiber.
	<ol>
		<li>Strip approximately 30 mm of the fiber with a fiber stripping tool. For fragile fibers (e.g., double clad fibers), a razor blade can be used to...

<ol>
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E., Hartl, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ultrafast+fibre+lasers.">Ultrafast fibre lasers.</a> Nature Photonics. 7, 868-874 (2013).</li><li>Szczepanek, J., Karda&#347;, T. M., Michalska, M., Radzewicz, C., Stepanenko, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Simple+all-PM-fiber+laser+mode-locked+with+a+nonlinear+loop+mirror.">Simple all-PM-fiber laser mode-locked with a nonlinear loop mirror.</a> Optics Letters. 40, 3500-3503 (2015).</li><li>Bowen, P., Singh, H., Runge, A., Provo, R., Broderick, N. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mode-locked+femtosecond+all-normal+all-PM+Yb-doped+fiber+laser+at+1060+nm.">Mode-locked femtosecond all-normal all-PM Yb-doped fiber laser at 1060 nm.</a> Optics Communications. 364, 181-184 (2016).</li><li>Chong, A., Buckley, J., Renninger, W., Wise, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=All-normal-dispersion+femtosecond+fiber+laser.">All-normal-dispersion femtosecond fiber laser.</a> Optics Express. 14, 10095-10100 (2006).</li><li>Kieu, K., Renninger, W., Chong, A., Wise, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sub-100+fs+pulses+at+watt-level+powers+from+a+dissipative-soliton+fiber+laser.">Sub-100 fs pulses at watt-level powers from a dissipative-soliton fiber laser.</a> Optics Letters. 34, 593-595 (2009).</li><li>Wise, F. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Femtosecond+Fiber+Lasers+Based+on+Dissipative+Processes+for+Nonlinear+Microscopy.">Femtosecond Fiber Lasers Based on Dissipative Processes for Nonlinear Microscopy.</a> IEEE Journal of Selected Topics in Quantum Electronics. 18, 1412-1421 (2012).</li><li>Nielsen, C. K., Keiding, S. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=All-fiber+mode-locked+fiber+laser.">All-fiber mode-locked fiber laser.</a> Optics Letters. 32, 1474 (2007).</li><li>Liu, Z., Ziegler, Z. M., Wright, L. G., Wise, F. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Megawatt+peak+power+from+a+Mamyshev+oscillator.">Megawatt peak power from a Mamyshev oscillator.</a> Optica. 4, 649-654 (2017).</li><li>Sidorenko, P., Fu, W., Wright, L. G., Olivier, M., Wise, F. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Self-seeded,+multi-megawatt,+Mamyshev+oscillator.">Self-seeded, multi-megawatt, Mamyshev oscillator.</a> Optics Letters. 43, 2672-2675 (2018).</li><li>Li, X., 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+ultrafast+Yb:fiber+laser+frequency+combs+using+commercially+available+components+and+basic+fiber+tools.">High-power ultrafast Yb:fiber laser frequency combs using commercially available components and basic fiber tools.</a> Review of Scientific Instruments. 87, 093114 (2016).</li><li>Bale, B., Kieu, K., Kutz, J., Wise, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transition+dynamics+for+multi-pulsing+in+mode-locked+lasers.">Transition dynamics for multi-pulsing in mode-locked lasers.</a> Optics Express. 17, 23137-23146 (2009).</li><li>Davoudzadeh, N., Ducourthial, G., Spring, B. Q. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Custom+fabrication+and+mode-locked+operation+of+a+femtosecond+fiber+laser+for+multiphoton+microscopy.">Custom fabrication and mode-locked operation of a femtosecond fiber laser for multiphoton microscopy.</a> Scientific Reports. 9, 4233 (2019).</li><li>Renninger, W., Chong, A., Wise, F. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Area+theorem+and+energy+quantization+for+dissipative+optical+solitons.">Area theorem and energy quantization for dissipative optical solitons.</a> Journal of the Optical Society of America. 27, 1978-1982 (2010).</li></ol>

The protocols outlined here synthesize know-how and expertise that have been common practice in the laser physics laboratory for decades, but which is frequently unfamiliar to many biomedical researchers. This work attempts to make this ultrafast fiber laser technology more accessible to the broader community. The ANDi fiber laser design is well-established, as first developed in seminal works by Wise and colleagues3. However, implementations of this technology by other groups have sometimes resul...

Solid state femtosecond (fs) pulsed lasers are widely used for microscopy and biological research. One typical example is the usage of multiphoton excitation (MPE) fluorescence microscopy, where high peak power and low average power are desired to facilitate the MPE process while minimizing photodamage mechanisms. Many high-performance solid-state lasers are commercially available, and when combined with an optical parametric oscillator (OPO), the laser wavelength can be tuned over a wide range1. For example, commercial oscillator-OPO systems generate &#60;120 fs pulse durations (typically with an 80 MHz pulse repetition rate) and &#62;1 W average power from 680 to 1,300 nm. However, the cost of these commercial tunable fs laser systems is significant (&#62;$200,000), and solid-state systems generally require water cooling and are not portable for clinical applications.
Ultrashort pulsed fiber laser technology has matured in the past few years. The cost of a commercial fs pulsed fiber laser is typically significantly lower than solid-state lasers, albeit without the capability of broad wavelength tuning afforded by the solid-state systems mentioned above. Note that fiber lasers can be paired with OPOs when desired (i.e., hybrid fiber-solid-state systems). The large surface-to-volume ratio of fiber laser systems enables efficient air cooling2. Hence, fiber lasers are more portable than solid-state systems due to their relatively small size and simplified cooling system. Further, fusion splicing of the fiber components reduces system complexity and mechanical drift in contrast to the free-space alignment of the optical components making up solid-state devices. All of these features make fiber lasers ideal for clinical applications. In fact, all-fiber lasers have been developed for low-maintenance operation3,4,5, and all-polarization-maintaining (PM)-fiber lasers are stable to environmental factors including changes in temperature and humidity as well as mechanical vibrations2,6,7,8.
Here, a method is presented to build a cost-efficient fs pulsed ANDi fiber laser with commercially available parts and standard fiber splicing techniques. Methods to characterize pulse repetition rate, duration, and coherence (full mode-lock) are also presented. The resulting fiber laser generates mode-locked pulses that can be compressed to 70 fs with a repetition rate of 31 MHz and a wavelength centered at 1,060 to 1,070 nm. The maximum power output from the laser cavity is approximately 1 W. The pulse physics of ANDi fiber lasers elegantly utilizes nonlinear polarization evolution intrinsic to optical fiber as a key component of the saturable absorber2,3,9,10,11. However, this means that the ANDi design is not easily implemented using PM fiber (although an all-PM fiber implementation of ANDi mode-locking has been reported, albeit with low power and ps pulse duration12). Thus, environmental stability requires significant engineering. Next generation fiber laser designs, such as the Mamyshev oscillator, have the potential to offer complete environmental stability as all-PM-fiber devices capable of an order-of-magnitude increase in intracavity pulse energy as well as offering significant decreases in pulse duration to enable applications that rely on broad pulse spectra13,14. Custom fabrication of these innovative new fs fiber laser designs requires know-how and fiber splicing experience.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Adapters, mirrors, posts, mounts, and translational stage (optomechanics)</td>
			<td>Thorlabs</td>
			<td>TR6-P5 (3x), AD12NT (2x), PFSQ20-03-M01, PFSQ05-03-M01, KMS, KM100C, KM100CL, KM200S, LT1, LT101, UPH2-P5, UPH3-P5 (2x)</td>
			<td>Standard optical components</td>
		</tr>
		<tr>
			<td>Advanced optical fiber cleaver</td>
			<td>AFL</td>
			<td>CT-100</td>
			<td></td>
		</tr>
		<tr>
			<td>Autocorrelator</td>
			<td>Femtochrome</td>
			<td>FR-103XL/IR/FA/CDA</td>
			<td></td>
		</tr>
		<tr>
			<td>Beamsplitter mount</td>
			<td>Thorlabs</td>
			<td>BSH1/M</td>
			<td></td>
		</tr>
		<tr>
			<td>Factory fusion splicer</td>
			<td>AFL</td>
			<td>FSM-100P</td>
			<td></td>
		</tr>
		<tr>
			<td>Fiber collimators</td>
			<td>OZ Optics (Canada)</td>
			<td>LPC-08-1064-6/125-S-1.6-7.5AS-60-X-1-2-HPC</td>
			<td>3x</td>
		</tr>
		<tr>
			<td>Fiber-coupled,high-speed photodiode detector</td>
			<td>Thorlabs</td>
			<td>DET08CFC</td>
			<td></td>
		</tr>
		<tr>
			<td>Free-space isolator</td>
			<td>Thorlabs</td>
			<td>IO-5-1050-HP</td>
			<td></td>
		</tr>
		<tr>
			<td>Free-space isolator</td>
			<td>Thorlabs</td>
			<td>IO-3D-1050-VLP</td>
			<td></td>
		</tr>
		<tr>
			<td>Half waveplate</td>
			<td>Union Optics (China)</td>
			<td>WPZ2312</td>
			<td>2x</td>
		</tr>
		<tr>
			<td>High power multimode fiber pump module</td>
			<td>Gauss Lasers (China)</td>
			<td>Pump-MM-976-10</td>
			<td></td>
		</tr>
		<tr>
			<td>High power pump and signal combiner</td>
			<td>ITF Technology (Canada)</td>
			<td>MMC02112DF1</td>
			<td></td>
		</tr>
		<tr>
			<td>Index matching gel</td>
			<td>Thorlabs</td>
			<td>G608N3</td>
			<td></td>
		</tr>
		<tr>
			<td>Optical spectrum analyzer</td>
			<td>Keysight</td>
			<td>Agilent 70951B</td>
			<td></td>
		</tr>
		<tr>
			<td>Oscilloscope</td>
			<td>Keysight</td>
			<td>Agilent 54845A</td>
			<td></td>
		</tr>
		<tr>
			<td>Passive double clad fiber(5/130 &#956;m)</td>
			<td>ITF Technology (Canada)</td>
			<td>MMC02112DF1</td>
			<td>3m, Included with combiner</td>
		</tr>
		<tr>
			<td>Polarizing beamsplitter</td>
			<td>Thorlabs</td>
			<td>PBS253</td>
			<td></td>
		</tr>
		<tr>
			<td>Quarter waveplates</td>
			<td>Union Optics (China)</td>
			<td>WPZ4312</td>
			<td>2x</td>
		</tr>
		<tr>
			<td>Quartz birefringent filter plate</td>
			<td>Newlight (Canada)</td>
			<td>BIR1060</td>
			<td></td>
		</tr>
		<tr>
			<td>RF spectrum analyzer</td>
			<td>Tektronix</td>
			<td>RSA306B</td>
			<td></td>
		</tr>
		<tr>
			<td>Single mode fiber (6/125 &#956;m)</td>
			<td>OZ Optics (Canada)</td>
			<td>LPC-08-1064-6/125-S-1.6-7.5AS-60-X-1-2-HPC</td>
			<td>1m, Included with collimators</td>
		</tr>
		<tr>
			<td>Single mode fiber coupler</td>
			<td>AFW (Australia)</td>
			<td>FOSC-2-64-30-L-1-H64-2</td>
			<td></td>
		</tr>
		<tr>
			<td>Transmission diffraction grating 1</td>
			<td>LightSmyth</td>
			<td>T-1000-1040-3212-94</td>
			<td>For compressor</td>
		</tr>
		<tr>
			<td>Transmission diffraction grating 2</td>
			<td>LightSmyth</td>
			<td>T-1000-1040-60x12.3-94</td>
			<td>For compressor</td>
		</tr>
		<tr>
			<td>Waveplate rotation mount</td>
			<td>Thorlabs</td>
			<td>RSP1/M</td>
			<td>4x</td>
		</tr>
		<tr>
			<td>Ytterbium-doped single mode double clad fiber</td>
			<td>Thorlabs</td>
			<td>YB1200-6/125DC</td>
			<td>3m</td>
		</tr>
	</tbody>
</table>

low-cost custom fabrication and mode-locked operation of an all-normal-dispersion femtosecond fiber laser for multiphoton microscopy

A protocol is presented to build a custom low-cost yet high-performance femtosecond (fs) fiber laser. This all-normal-dispersion (ANDi) ytterbium-doped fiber laser is built completely using commercially available parts, including $8,000 in fiber optic and pump laser components, plus $4,800 in standard optical components and extra-cavity accessories. Researchers new to fiber optic device fabrication may also consider investing in basic fiber splicing and laser pulse characterization equipment (~$63,000). Important for optimal laser operation, methods to verify true versus apparent (partial or noise-like) mode-locked performance are presented. This system achieves 70 fs pulse duration with a center wavelength of approximately 1,070 nm and a pulse repetition rate of 31 MHz. This fiber laser exhibits the peak performance that may be obtained for an easily assembled fiber laser system, which makes this design ideal for research laboratories aiming to develop compact and portable fs laser technologies that enable new implementations of clinical multiphoton microscopy and fs surgery.

It is critical to verify mode-locked operation upon completion of the fiber laser fabrication procedures. Signatures of optimal fs pulse generation and laser stability are as follows: First, the output pulse may be sufficiently characterized by the instrumentation outlined in step 6. The pulse spectrum output from the laser oscillator should be centered near 1,070 nm with the characteristic cat-ear or Batman shape that indicates mode-locking as predicted by numerical simulation of ANDi pu...

Watch this Scientific Journal Video about Low-cost Custom Fabrication and Mode-locked Operation of an All-normal-dispersion Femtosecond Fiber Laser for Multiphoton Microscopy at JoVE.com

Low-cost Custom Fabrication and Mode-locked Operation of an All-normal-dispersion Femtosecond Fiber Laser for Multiphoton Microscopy

A method is presented to build a custom low-cost, mode-locked femtosecond fiber laser for potential applications in multiphoton microscopy, endoscopy, and photomedicine. This laser is built using commercially available parts and basic splicing techniques.

A protocol is presented to build a custom low-cost yet high-performance femtosecond (fs) fiber laser. This all-normal-dispersion (ANDi) ytterbium-doped ...

Femtosecond pulse lasers have broad applications in multiphoton miscroscopy. This protocol can be used to fabricate a femtosecond all-normal dispersion fiber laser that is compact, robust, and inexpensive. Compared with commercial solid-state ultrafast lasers, the laser produced in this technique costs much less because it consists of only commercially available parts.Also, fiber lasers do not need water cooling, so the size of the system is smaller. Last but not least, the fiber components do not require alignment, which makes the system robust to vibration. Unlike commercially available systems, this laser does not have a cover to block unwanted beams.Experienced personnel are needed to assemble and operate the laser. Some experiments appear to be unreproducable because it is very likely to miss some uncalled details when following written instructions. In video demonstration, the viewers won't miss anything.Start by splicing single-mode fibers, or SMFs, in order to ensure proper performance of the splicing equipment before more valuable fiber optic materials are used. Strip approximately 30 millimeters of the fiber with a fiber-stripping tool. If working with fragile fibers, a razor blade can be used to carefully peel off the buffer.Use a lint-free tissue with ethanol or isopropanol to clean the stripped fiber. A buzzing sound while wiping indicates that the fiber is sufficiently clean. Then, place the fiber holder on the fiber cleaver and make sure that the blade, fiber clamp of the cleaver, and the fiber holder are all clean.Carefully load the fiber into the fiber holder leaving approximately 25 millimeters of stripped clean fiber at the free end of the cleaver to clamp. Gently close the fiber clamp on the cleaver. To avoid excess tension on the fiber, reopen and close the clamp.Press the Cut button and the cleaver will automatically cut the fiber. Use tweezers with plastic-rounded tips to move the piece cut out from the fiber to a sharps disposal container and transfer the fiber holder to the fusion splicer. Repeat the procedure to cleave the second fiber.The two fibers to be spliced together should have cleaved ends opposing each other by the fiber holders within the fiber splicer. Close the cover of the splicer and set parameters such as Core Diameter, Mode Field Diameter, and Cladding Diameter. Set the alignment method to Cladding, press the Set button, and the splicer will align automatically.Press the Set button at every stop to confirm the quality of the alignment. The splice will be done automatically. Check the quality of the splice using the quality controls of the splicer as well as the camera view of the region.A good splice has a uniform cladding boundary and uniform brightness along the fiber such that no splice juncture is visible. Then, open the splicer cover and one of the fiber holders. Optionally, a fiber sleeve may be added to protect the splice and the heater of the splicer can be used to mold the sleeve onto the fiber.Splice the combiner output to the ytterbium-doped active fiber. Follow the previously described procedure to cleave the combiner output fiber. Due to the shape of its cladding, the active fiber should first be cleaved and spliced with a piece of single-mode fiber which will later be removed.Cut the single-mode fiber about two centimeters from the splicing point with a wirecutter. Then, strip the entire length of the single-mode fiber and 0.5 centimeters of the active fiber which will leave the active fiber capped with two centimeters of bufferless single-mode fiber. Load the active fiber into the cleaver making sure that only the single-mode fiber is clamped by the fiber clamp.From this point on, follow the previously described procedure for cleaving and splicing the fiber. Safety is the top priority. Remember to put every piece of fiber fragment in the sharp object box.Also, laser safety goggles should be warn whenever the pump is operating. Turn on the oscilloscope and set the instrument to AC coupling mode with the trigger level set to 30 millivolts. Move the optical spectrum analyzer photodiode input fiber to monochromatic input and set the device to OSA mode.Then, lock the phase of the laser by adjusting the wave plates. Rotate Quarter-Wave Plate 2 several degrees back and forth. The mode-locking spectrum consists of two stable peaks with a plateau between them.Meanwhile, observe a stable pulse train on the oscilloscope. If the mode-locking spectrum is not observed, rotate Quarter-Wave Plate 1 several degrees in one direction and repeat the previous step. If the spectrum is still not observed, rotate the birefringent filter several degrees and repeat the process.Mode-locked operation was verified upon completion of fiber laser fabrication. The pulse spectrum output from the laser oscillator was centered near 1070 nanometers with the characteristic cat ear shape which indicates mode-locking as predicted by numerical simulation. As a further diagnostic for mode-locking, the pulse duration and pulse repetition power spectra were measured using the autocorrelator and radio frequency spectrum analyzer respectively.Pulse durations of 70 femtoseconds were measured. Pulse stability was tested by continuously monitoring the average output power and the pulse spectrum. When the laser setup was mounted on a floating optical table with vibration damping, the power drift was less than 3.5%over 24 hours without active cooling.After verifying mode-locking, the imaging performance was tested using simple test targets and biological samples. Fluorescence was measured during adjustments of the pulse power which verified that the signal was quadratically dependent on the laser power delivered to the sample plane. Stained and unstained biological specimens were imaged using the custom-built fiber laser.As an additional verification of two-photon excitation, collected hyperspectral images of multicolor fluorescent microspheres were compared with images taken by linear excitation with commercial diode lasers. Finally, the normalized spectra of green and red beads excited by the diode laser versus the custom FS fiber laser were compared. The free space component can be replaced by corresponding fiber parts which can further increase robustness and mobility.The all-fiber system can be put on a cart for clinical scenarios. The free space component can be replaced by corresponding fiber parts which can further increase robustness and mobility. The all-fiber system can be put on a cart for clinical scenarios.The impact of this technology is an open question. We anticipate that it will give researchers new access to femtosecond laser technology and enable them to develop new publications.

Research

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