The authors gratefully acknowledge MBI financial support.

1. Preparation of Silicon Primary Mold
NOTE: A photomask is required for the preparation of the primary mold. The photomask can be either purchased from independent providers or fabricated in-house through standard optical mask fabrication procedures. A photomask with bright field polarity is used in this case (Figure 2a).
<ol>
	<li>Pattern definition
	<ol>
		<li>Spin coat a 4 inch silicon wafer with SU-8 3010 negative photoresist at 2,300 rpm for 30 s.</li>
		<li>Soft-bake the photoresist on a hot plate at 65 &#176;C for 2 min and then at 95 &#176;C for 8 min.</li>
		<li>Expose the photoresist to UV light (i-line, 365 nm) through the photomask under the mask aligner for a total energy dose of 100-120 mJ/cm2; the hard contact mode is preferred to achieve a better resolution.</li>
		<li>Remove the wafer and apply a post exposure bake at 65 &#176;C for 1 min and then at 95 &#176;C for 2 min.</li>
		<li>Develop the photoresist using a SU-8 developer at room temperature, then rinse with isopropyl alcohol, and gently blow dry with nitrogen; the measured pattern thickness should be 10 &#181;m and below. See Figure 2b for the picture of the actual silicon mold.</li>
	</ol>
	</li>
	<li>Silanization of silicon mold
	<ol>
		<li>Treat the silicon mold with oxygen plasma at 60 W for 30 s with 20 sccm of oxygen flow. Set the chamber pressure to 1-10 mbar during the process.</li>
		<li>Place the mold in a vacuum jar with 50 &#181;L of silane and leave the jar in the vacuum state (1-10 mbar) for at least 2 h. 
		NOTE: The silanization process creates a hydrophobic surface coating which prevents PDMS from sticking to the mold15. Note that the primary mold can also be fabricated using an alternative method, which involves dry etching of the silicon. In this case, the photomask will be of opposite polarity (dark field), and the pattern definition in step 1.1 will use a positive photoresist.</li>
	</ol>
	</li>
</ol>
2. Preparation of PDMS Secondary Mold
<ol>
	<li>PDMS mixing
	<ol>
		<li>Mix the PDMS elastomer and curing agent, 10:1; the total amount is such that the resulting PDMS thickness is approximately 1 to 1.5 mm.</li>
		<li>After thorough mixing, degas the mixture by leaving it in a vacuum jar in the vacuum state (1-10 mbar) for approximately 15 min or until there are no visible bubbles; this is to remove any trapped air within the mixture.</li>
	</ol>
	</li>
	<li>Mold replication
	<ol>
		<li>Pour the PDMS mixture on the silicon mold fabricated in step 1 and degas the mixture to remove any trapped air with the same settings as in step 2.1.2. Heat at 70 &#176;C for 2 h on a hot plate to cure the mixture.</li>
		<li>Remove the cured PDMS from the hot plate and let it cool down to room temperature. With a razor blade, cut the PDMS along the edges of the silicon mold.</li>
		<li>With a pair of tweezers, pinch one corner of the cut PDMS and carefully peel the PDMS replica off the silicon mold; the resulting microfluidic pattern on this secondary mold is a protrusion, which is the opposite polarity of the primary mold (Figure 2c).</li>
	</ol>
	</li>
	<li>Silanization of the PDMS replica (same as in step 1.2)
	<ol>
		<li>Treat the PDMS mold with oxygen plasma at 60 W for 30 s with 20 sccm of oxygen flow. Set the chamber pressure to 1-10 mbar during the process.</li>
		<li>Place the mold in a vacuum jar with 50 &#181;L of silane and leave the jar in the vacuum state (1-10 mbar) for at least 2 h.</li>
	</ol>
	</li>
</ol>
3. Preparation of PDMS Templates
NOTE: To standardize the shape and size of the final devices and to ease the alignment of the main features in the two halves, two separate PDMS templates were used, which define the geometry of the device, the placement of the transparent window and the in- and out-let connections. The first PDMS template aids the fabrication of the patterned half of the device, while the second helps to ease the fabrication of the flat half of the device.
<ol>
	<li>Design the templates using a computer aided design (CAD) software. Figure 3a shows the lay-out and dimensions of the template used to fabricate the patterned half of the device. To fabricate the flat half of the device, remove the 1.5 mm diameter holes from the design.</li>
	<li>Acquire the templates from an external provider or through an internal mechanical workshop if available. 
	NOTE: Acrylic was used as the template material due to ease of fabrication and low cost achievable in any standard workshop. There are alternative options, such as 3D printing.</li>
	<li>Mix the PDMS elastomer and curing agent 10:1; be sure to prepare a sufficient amount of PDMS to entirely submerge the templates.</li>
	<li>After thorough mixing, degas the mixture by leaving it in a vacuum jar in the vacuum state (1-10 mbar) for approximately 15 min or until there are no visible bubbles (whichever is later); this is to remove any trapped air within the mixture.</li>
	<li>Pour the PDMS mixture on the acrylic templates until their top-most surface is submerged about 1 mm below the liquid surface. Degas the PDMS again to remove any trapped air with the same settings as in 3.4. Heat this at 60 &#176;C for 2 h on a leveled hot plate to cure the mixture.</li>
	<li>Remove the cured PDMS from the hot plate and let it cool down to room temperature. With a razor blade, cut the PDMS along the edges of the acrylic templates.</li>
	<li>With a pair of tweezers, pinch one corner of the cut PDMS and carefully peel the PDMS off the acrylic templates. 
	NOTE: Figure 3b shows the lay-out and dimensions of the PDMS replica used to fabricate the patterned half of the device.</li>
	<li>Prepare the second PDMS replica for fabricating the flat half of the device by repeating steps 3.3 to 3.7 but using the acrylic template without the 1.5 mm diameter holes.</li>
</ol>
4. Microfluidic Device Fabrication
<ol>
	<li>Fabrication of the patterned half of the device (i.e. , with device layout)
	<ol>
		<li>Treat the CaF2 window with oxygen plasma at 60 W for 30 s with 20 sccm of oxygen flow. This is done to improve the flow of NOA during the following fabrication. 
		NOTE: This step is not mandatory.</li>
		<li>Place carefully the first PDMS template (one with the 1.5 mm diameter pillars) on a flat surface, e.g., a soda lime plate (Figure 4a). Place a CaF2 window centered on top of the PDMS plug and gently press the window such that it is in good contact with the plug (Figure 4b).</li>
		<li>Take the PDMS mold made in step 2 and place a thin UV-transparent plate (in this case, a quartz plate, 500 &#181;m thick and 1.5 cm x 1.5 cm in size) on the backside of the mold, aligned with the location of the central chamber (Figure 4c). Make sure that the quartz plate is in good contact with the PDMS mold. 
		NOTE: The quartz plate prevents the unwanted area of the mold from easily coming into contact with the CaF2 window.</li>
		<li>Gently place this PDMS mold face down towards the CaF2 window with the fluidic chamber aligned to the center of the CaF2 window. Make sure that all the elements (template, mold and window) are in good contact and aligned (Figure 4c-4d).</li>
		<li>Gradually dispense drops of NOA at the in-let of the PDMS template and let it slowly fill the cavity. Once the resin comes into contact with the edge of the window, the capillary flow will fill the thin gap (~10 &#181;m) between the PDMS mold and CaF2 window (Figure 4e-4f).</li>
		<li>After the cavity is completely filled, cure the NOA by exposure to UV light (e.g., with a UV-LED exposure system, Figure 4g). 
		NOTE: The exposure time may vary accordingly with the energy of the UV source. The UV-LED exposure system, which provides a power density of 24 mW/cm2, requires around 90 s at 100% power and continuous exposure mode.</li>
		<li>Carefully remove the thin quartz plate from the back of the PDMS mold and then gently peel the PDMS mold from the top of the NOA layer (Figure 4h). Finally, remove the NOA layer from the PDMS template (Figure 4i). 
		NOTE: The resulting device layout on the cured NOA would have the same polarity of the pattern in the primary silicon mold.</li>
	</ol>
	</li>
	<li>Fabrication of the flat half of the device (i.e. , without device layout)
	<ol>
		<li>Treat the CaF2 window with oxygen plasma at 60 W for 30 s with 20 sccm of oxygen flow. 
		NOTE: This step is not mandatory.</li>
		<li>Place carefully the second PDMS template (one without the 1.5 mm diameter pillars) on a flat surface, e.g., a soda lime plate. Place a CaF2 window centered on top of the PDMS plug and gently press the window such that it is in good contact with the plug.</li>
		<li>Place a 1 mm thick PDMS sheet with 5 cm x 3.5 cm size on top of the CaF2 window, with the PDMS sheet aligned with the center of the PDMS template. Make sure that the PDMS sheet is in good contact with the window.</li>
		<li>Gradually dispense drops of NOA at the in-let of the PDMS template and let it slowly fill the cavity.</li>
		<li>After the cavity is completely filled, cure the NOA by exposure to UV light (e.g., with a UV-LED exposure system). 
		NOTE: The exposure time may vary accordingly with the UV energy source. With the UV-LED exposure system, which provides a power density of 24 mW/cm2, this requires around 50 s at 100% power and continuous exposure mode.</li>
		<li>Peel off the PDMS sheet from the top of the NOA layer and carefully remove the cured NOA layer from the PDMS template.</li>
	</ol>
	</li>
	<li>Bonding of the two halves of the device
	<ol>
		<li>Align the two halves of the device such that both CaF2 windows are aligned. Gently finger-press both halves at the corner of the NOA layers such that the position of the two halves are fixed.</li>
		<li>Cut two circular discs out of a 1 mm thick PDMS sheet using an 8 mm diameter puncher (Figure 5a).</li>
		<li>Cut two rectangles with the same size of the device (4 cm x 2.5 cm) from a 1 mm thick PDMS sheet. On both PDMS rectangles, cut openings corresponding to the channels and the in-let/out-let of the device. 
		NOTE: The pre-cut openings in the PDMS rectangles are meant to prevent the channels from collapsing during pressing.</li>
		<li>Stack in the following order from the bottom: one PDMS rectangle with pre-cut openings, one PDMS disc (in contact with the bottom window, cut in step 4.3.2), the two finger-pressed halves of the device, the second PDMS disc (sitting on the top window), and lastly the second PDMS rectangle with pre-cut openings (Figure 5b).</li>
		<li>Place this assembly in the vacuum press setup such that it is sandwiched between 2 plates and seal the plastic bag (Figure 5c). Turn on the vacuum pump and evacuate the assembly. Let the vacuum pump reach its base pressure or apply the vacuum for at least 10 min. 
		NOTE: The base pressure achieved depends on the vacuum pump used and quality of the sealing of the plastic bag.</li>
		<li>Expose the evacuated assembly to UV with a broad-band Hg gas lamp at 270 W for 15 min. Turn off the vacuum pump and let the assembly slowly vent to atmospheric pressure before removing the final device from the assembly.</li>
	</ol>
	</li>
</ol>

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A., Keese, M., Pepperkok, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=IR+microspectroscopy+of+live+cells.">IR microspectroscopy of live cells.</a> Vibrational Spectroscopy. 38 (1-2), 185-191 (2005).</li><li>Rahmelow, K., Hubner, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Infrared+Spectroscopy+in+Aqueous+Solution:+Difficulties+and+Accuracy+of+Water+Subtraction.">Infrared Spectroscopy in Aqueous Solution: Difficulties and Accuracy of Water Subtraction.</a> Appl Spectrosc. 51 (2), 160-170 (1997).</li><li>Kazarian, S. G., Chan, K. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=ATR-FTIR+spectroscopic+imaging:+recent+advances+and+applications+to+biological+systems.">ATR-FTIR spectroscopic imaging: recent advances and applications to biological systems.</a> Analyst. 138 (7), 1940-1951 (2013).</li><li>Loutherback, K., Chen, L., Holman, H. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Open-channel+microfluidic+membrane+device+for+long-term+FT-IR+spectromicroscopy+of+live+adherent+cells.">Open-channel microfluidic membrane device for long-term FT-IR spectromicroscopy of live adherent cells.</a> Anal Chem. 87 (9), 4601-4606 (2015).</li><li>Loutherback, K., Birarda, G., Chen, L., Holman, H. -. 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The authors have nothing to disclose.

In order to assess and optimize the fabrication protocol, we used a simple layout for the microfluidic pattern with a large rectangular chamber (5 mm x 2.5 mm size) at the center, two small rectangular chambers (5.5 mm x 0.75 mm size) separated from the main circuit on the upper and lower sides, and 300 &#181;m wide in-let/out-let channels. The central chamber is used for the seeding and observation of the cells, while the two separated smaller chambers are used to measure the air background during FTIR experiments as a reference chamber, as discussed in a previous publication13. The in-let and out-let connect the central chamber to the external fluidic system.
Here, templating tools (see protocol step 3) are introduced to ease the fabrication process. Previously, it was difficult to consistently place the CaF2 windows exactly at the center of the final device, which caused issues when aligning the two halves of the device. The use of the template provides a visual guide for placing the window and ensures that the location is always similar. As the geometrical requirements for these templates are not very stringent, standard and cheap production technologies can be used. In this case, we made them out of acrylic plastic through machining in a workshop, but an equally viable and cheap alternative is 3D printing.
The design of the acrylic templates is such that they feature the following (Figure 3a): (a) one template is designed with two small holes on each side, while the second is without holes; the holes are 1.5 mm diameter in size and 1.5 mm in depth, (b) both templates have a circular hole with 8 mm diameter and 500 &#181;m depth in the center, and (c) both templates have a rectangular protrusion to define the size, shape, and thickness of the half device; the rectangle is 4 cm x 2.5 cm in length/width and 1.5 mm in thickness.
The resulting PDMS templates replicated from the acrylic templates will have the opposite polarity (Figure 3b): (a) one template will have two pillars on each side with 1.5 mm diameter and 1.5 mm height to form the in-let and out-let of the patterned half of the device after NOA casting; the second template will be without pillars, (b) both templates have a 500 &#181;m tall pillar in the center to ease the placement of the CaF2 windows; this feature will be referred to as &#34;PDMS plug&#34;, and (c) both templates are with a cavity, which defines the final shape and size of each half of the device: a rectangle shape with 4 cm x 2.5 cm size and 1.5 mm thickness in this case.
To further ease the fabrication process, diffraction gratings were incorporated in the design of the photomask for the primary silicon mold. In the PDMS replica of this silicon mold, the shallow protrusions (which are less than 10 &#181;m in height) are hard to see due to the optical transparency of PDMS. However, shallow protrusions if arranged in a grating of 10 &#181;m wide lines with 20 &#181;m gap generate an easily visible interference pattern16. This interference pattern was exploited as a visual guide to define the position of the microfluidic lay-out in the PDMS mold. The design of the mold incorporates a frame made of the grating to define the overall geometry of the device. Another grating was further added in the middle of the central chamber of the microfluidic layout in order to make alignment with the CaF2 window easier. It is worth noting that the grating in the central chamber will not be reproduced in the final device as it consists of a series of short cavities which are not connected to the edge of the central chamber. Therefore, the flowing NOA would not be able to access these cavities.
For the cell injection and medium exchange, every device needs its in-let and out-let manually punched before sealing. This resulted in non-reproducible positioning of the holes. This did not limit the operation of each device since the connection was achieved by gluing a small metallic pin at one of the openings and attaching a plastic reservoir at the other end as a waste collector. Therefore, varying positions of the holes is not of concern except at the cost of a more complex fabrication scheme. To simplify and to standardize the testing and final usage of every device, a custom-made fluidic jig with fixed in-let/out-let locations is introduced (see Figure 8), removing the need to attach the pin and reservoir. Hence, in-let/out-let holes with inconsistent positions would present an issue here. By incorporating two 1.5 mm diameter holes in the fabricated acrylic template (protocol step 3), which will become pillars in the PDMS mold, the need to manually punch holes at the ends of the microfluidic channels is removed. Moreover, their position is fixed and the same for every device.
The absence of leaks in the device can be checked with the same procedure as discussed elsewhere13 i.e., by feeding the device with a solution of fluorescein in de-ionized water.
Critical steps in the protocol 
During fabrication of the patterned half of the device, placement of the PDMS mold on top of the CaF2 window should be carefully carried out. The only structures on the mold which are allowed to contact the window are the 10 &#181;m high protrusions. Whenever there is an unwanted contact area, the mold placement step should be redone. To illustrate, Figure 9a shows careful placement of the mold on the window, while Figure 9b shows an example of poor placement, where an unwanted area of the mold is in contact with the window. Figure 9b would result in a missing NOA structure on the window. The small quartz plate mentioned in the protocol (step 4.1.3, Figure 2c) helps the mold placement as it prevents the unwanted area of the mold from easily coming into contact with the CaF2 window. Both the quartz plate and PDMS mold are also thin so that they are sufficiently light-weight and transparent to UV17,18.
Finding the correct UV exposure dose for the half layers is also crucial for the fabrication process. When NOA is exposed with an insufficient dose, the uncured NOA will reflow during the peeling off, causing a loss of definition of the structure and possibly overflowing onto the CaF2 surface. On the other hand, too high of an exposure dose results in over-curing of the NOA, turning NOA into a non-tacky state. Subsequent bonding of the two halves would suffer because non-tacky NOA on the patterned half layer would not bond to the CaF2 window on the flat half layer. Ideally, the correct dose should be the shortest exposure dose which allows NOA to replicate the structure reliably while still maintaining its tacky state. Besides determining the correct exposure dose, bonding of the two halves should be done as soon as possible, within 30 min, as the stickiness of NOA gradually decreases with time until it is no longer possible for bonding.
Other points to note are when using the vacuum press system for bonding. The compliance layers (i.e., thin PDMS sheets sandwiching the device) should have relatively uniform thickness for uniform pressure distribution on the device. For this purpose, a custom-made acrylic jig with a defined spacer thickness was used to cast such PDMS sheets. The compliance sheets should also be clean to avoid introducing local pressure, especially on the brittle CaF2 window.
Advantages of the method compared to existing methods 
Our fabrication approach demonstrated the production of a plastic device compatible with FTIR measurements. Because the micro-fabrication technique provides good control over the dimension of the feature, the achievable control over the height of the microfluidic channels is much more precise than what can be obtained with other fabrication approaches (e.g., plastic spacers).
A crucial advantage of this protocol is that it results in a plastic device with UV-Vis-IR transparent view-ports; all the previously demonstrated microfluidic devices for FTIR were produced on top of a large IR transparent substrate, requiring lithography steps for each and every device10,11,12. In the present approach, the cost and complexity of the fabrication is reduced since only the production of the Si mold requires lithography.
Finally, using a UV curable resin (NOA73 in this demonstration) as the main plastic body eases the connectivity of the device to an external liquid delivery system, either by gluing or attaching the connections to the plastic body or by using a fluidic jig for more rapid device production or usage.
Future applications of the method 
Of the possible improvements and development that can be explored, two are the most immediate and important. First, the broadband optical transparency of the view-port suggests the coupling of FTIR with high-resolution fluorescence microscopy on the same platform. This can be easily pursued by reducing the thickness of one of the CaF2 windows in order to comply with the requirements of a working distance with high numerical aperture and high magnification objectives. Second, this fabrication scheme can allow for more complex fluidic layouts. Multiple observation chambers and functional elements connecting them such as mixers and sorters, can be implemented as long as their geometry defines the open cavities below the windows.

Fourier Transform Infrared Spectro-microscopy (FTIR) has been extensively utilized as a label-free and non-invasive imaging technique to provide detailed chemical information of a sample. This enables the extraction of biochemical information to study the chemistry of biological samples, with a minimum amount of preparation since the absorption spectrum of the specimen carries the intrinsic fingerprints of its chemical composition1,2. Recently, FTIR has been increasingly applied to the study of live biological samples, e.g., cells3. However, water, which is the medium for living cells in most cases, shows a strong absorbance in the mid-IR region. Even as a thin layer, its presence can completely overwhelm the important structural information of the specimens.
For many years, the common approach was fixing or drying samples to completely exclude the water absorption signal in the spectrum. However, this approach does not allow for real time measurements on living cells, which is essential to study the change of their chemical composition and cellular processes with time. One way to obtain reliable absorption spectra from live biological samples, is to limit the total optical path length in the medium of the IR beam to less than 10 &#181;m4.
A well-established approach in living cell experiments has been so far, Attenuated Total Reflection (ATR)-FTIR imaging, which enables measurements independent of the sample thickness, allowing cells to be sustained in a thicker layer of aqueous medium. However, the small depth of penetration of the evanescent wave restricts measurements of samples to only the first few microns from the surface of the ATR crystal5.
Alternatively, the water absorption limitation has been circumvented with the emergence of various microfluidic systems, which are generally classified into two large groups: open channel (where one of the fluid surfaces is exposed to the atmosphere) and closed channel (where two IR-transparent windows are separated by a spacer with a defined thickness).
Loutherback et al. developed an open-channel membrane device that enables long term continuous IR measurements of live cells for up to 7 days6. The method requires high humidity in the environment to prevent evaporation of the medium from the cell surface. The system works best with cells that naturally grow at air-liquid interfaces, such as epithelial tissues of the skin, lung, and eyes, or microbial biofilms7.
A closed-channel configuration aims to create a uniform, thin layer between two parallel IR-transparent windows, where cells are maintained in their aqueous media. The thickness of this cavity is such that the water absorption signal is below saturation. Water background can then be subtracted to obtain the correct sample spectra. Most of the closed-channel methods utilize a plastic spacer separating the two windows to form a demountable liquid chamber3,8,9. An advantage of this method is that it does not require microfabrication; however, structures that are more complex than a measuring chamber with in- and out-let channels are extremely difficult to realize in the thin spacer. There is also an issue with the reproducibility of the path length between IR measurements due to its reliance on mechanical clamping. In order to achieve a more precise control of the spacing for a more reliable spectrum acquisition, optical lithography methods have been implemented to pattern photoresist on top of the IR substrate to define the spacer9,10,11,12. Even though this makes it possible for more complex structures to be defined in the spacer, the method requires access to a microfabrication facility to produce the pattern on every substrate.
In this paper, we present a simple fabrication technique of an IR-compatible microfluidic device, with the aim to reduce the fabrication cost and the requirement of accessing a microfabrication facility. The method presented here (see Figure 1) uses an established process known as soft lithography. Two molds are required in this case. The primary mold is made from a 4-inch silicon wafer using a standard UV lithography process. The secondary mold is its replica made of PDMS, which has a reversed polarity of the pattern in the silicon primary mold and serves as the master mold for subsequent device fabrication.
The device has two separate layers: a first layer with the microfluidic layout (which in the presented case consists of the microfluidic channel, in-let/out-let, and an observation chamber with a CaF2 viewport), and a second layer with a flat surface (which consists of only a CaF2 viewport).
Here an UV-curable optical adhesive, Norland optical adhesive 73 (NOA73, abbreviated as NOA henceforth), is used to form the main plastic body of the device. There are several advantages of using this optical adhesive: low fabrication cost, ease of connectivity to external systems, good optical transparency, low viscosity, and most importantly, biocompatibility13. CaF2 is a suitable choice as the viewport due to its biocompatibility and excellent IR-transparency14.
With this new approach, access to a microfabrication facility is strictly required only for the fabrication of the primary mold. Subsequent fabrication processes for the plastic microfluidic device can be carried out in any laboratory equipped with an UV-illumination source.

<table><tbody><tr><td>&lt;strong&gt;Chemical&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>Trichloro(1H,1H,2H,2H-perfluorooctyl)silane 97%</td><td>Sigma Aldrich</td><td>448931-10G</td><td></td></tr><tr><td>Sylgard 184 Silicone Elastomer Kit</td><td>Dow Corning</td><td></td><td>Polydimethylsiloxane or in short, PDMS</td></tr><tr><td>Norland Optical Adhesive 73</td><td>Norland Products Inc.</td><td>7304</td><td></td></tr><tr><td>SU8 3010 photoresist</td><td>MicroChem</td><td>Y311060</td><td></td></tr><tr><td>SU8 developer</td><td>MicroChem</td><td>Y020100</td><td></td></tr><tr><td>&lt;strong&gt;Material&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>Silicon wafer, 4 inch, prime grade</td><td>Bonda Technology Pte Ltd</td><td></td><td></td></tr><tr><td>CaF2 IR-grade windows</td><td>Crystran, UK</td><td>CAFP10-1</td><td>10 mm diameter, 1 mm thickness</td></tr><tr><td>Acrylic templates</td><td></td><td></td><td>Custom made</td></tr><tr><td>&lt;strong&gt;Equipment&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>UV-KUB 2 (UV LED exposure system)</td><td>KLOE</td><td></td><td>Emission spectrum 365nm &amp;plusmn; 5nm</td></tr><tr><td>Newport UV lamp</td><td>Newport</td><td>Model 66902</td><td>50-500 Watt Hg arc lamp</td></tr><tr><td>CEE Spin coater</td><td>Brewer Science</td><td>Model 200x</td><td></td></tr><tr><td>MJB4 mask aligner</td><td>SUSS MicroTec</td><td></td><td></td></tr><tr><td>Precision digital hot plate</td><td>Harry Gestigkeit GmbH</td><td>2860SR</td><td></td></tr><tr><td>Plasma Surface Technology</td><td>Diener Electronic GmbH + Co. KG</td><td></td><td>For O&lt;sub&gt;2&lt;/sub&gt; plasma treatment</td></tr><tr><td>IDP-3 Dry Scroll Vacuum Pump</td><td>Agilent Technologies</td><td></td><td>ultimate pressure 3.3 x 10&lt;sup&gt;-1&lt;/sup&gt; mbar</td></tr><tr><td>Bruker IFS 66v/s FTIR Spectrometer</td><td>Bruker</td><td></td><td></td></tr></tbody></table>

soft lithographic procedure for producing plastic microfluidic devices with view-ports transparent to visible and infrared light

Infrared (IR) spectro-microscopy of living biological samples is hampered by the absorption of water in the mid-IR range and by the lack of suitable microfluidic devices. Here, a protocol for the fabrication of plastic microfluidic devices is demonstrated, where soft lithographic techniques are used to embed transparent Calcium Fluoride (CaF2) view-ports in connection with observation chamber(s). The method is based on a replica casting approach, where a polydimethylsiloxane (PDMS) mold is produced through standard lithographic procedures and then used as the template to produce a plastic device. The plastic device features ultraviolet/visible/infrared (UV/Vis/IR) -transparent windows made of CaF2 to allow for direct observation with visible and IR light. The advantages of the proposed method include: a reduced need for accessing a clean room micro-fabrication facility, multiple view-ports, an easy and versatile connection to an external pumping system through the plastic body, flexibility of the design, e.g., open/closed channels configuration, and the possibility to add sophisticated features such as nanoporous membranes.

Figure 6 presents the transmittance spectra of a brand new CaF2 window, the patterned half of the device, and the complete device. All three spectra exhibit excellent transparency to mid IR with transmittance larger than 80%. The interference pattern visible in the spectrum of the full device (yellow curve in the figure) is caused by the air gap in the range of 9-10 &#181;m between the two windows. These spectra demonstrate that the fabrication approach presented here does not alter the transparency of CaF2 in the mid IR range.
Figure 7a shows an example of good replication in NOA of the PDMS secondary mold with the microfluidic lay-out. The structure on top of the CaF2 window is well formed because of the clean peeling of the NOA layer from the PDMS mold after it is partially UV-cured. No NOA should remain on the mold or on the window surface in contact with the mold protrusions. Any NOA stuck to the mold translates into missing NOA structure on the window, which would cause leaks during flow experiments of the final device. In addition, to achieve good sealing of the two halves, NOA should still be tacky after UV exposure of the half layers. NOA is over-cured if it is non-tacky. The exposure dose should be optimized to achieve such outcome.
Figure 7b instead, shows unsuccessful replication in NOA where the pattern on the CaF2 window is not properly defined. It is mostly caused by insufficient UV exposure dose, i.e., under-curing of NOA. In such cases, NOA is still somewhat wet, causing some of it to stick to the PDMS mold. However, if NOA still sticks to PDMS mold even though the correct exposure dose has been given, this can be a symptom of silane coating (i.e., anti-stick layer) being degraded over time. As PDMS is a soft mold, it is not as long lasting as the silicon primary mold. It needs to be replaced after a number of uses.
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/55884/55884fig1.jpg" /> 
Figure 1: Fabrication process of the plastic microfluidic devices: (a-e) Schematic of the fabrication process. (f) Picture of an actual device and schematic of its cross-section. <a href="https://cloudflare-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/55884/55884fig1large.jpg" target="_blank">Please click here to view a larger version of this figure.</a>
<img alt="Figure 2" class="xfigimg" src="/files/ftp_upload/55884/55884fig2.jpg" /> 
Figure 2: Overview of the photomask, silicon primary mold, and PDMS secondary mold replicated from the silicon mold: (a) Photomask with bright field polarity (top left); Lay-out of the pattern in the photomask (right) consisting of: in-let and out-let with 2 mm diameters (1 and 2) and distance of 3 cm between them, channels with 300 &#181;m width (3), two reference chambers with 5.5 mm x 0.75 mm size (4), central chamber with 5 mm x 2.5 mm size (5), diffraction gratings as a visual guide with lines 10 &#181;m wide and 20 &#181;m gap (6); Zoom-in lay-out of the central chamber showing the diffraction grating (bottom left). (b) Picture of the silicon primary mold with the pattern defined in the SU-8 photoresist. (c) Picture of the PDMS secondary mold with reverse polarity with respect to the primary mold. <a href="https://cloudflare-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/55884/55884fig2large.jpg" target="_blank">Please click here to view a larger version of this figure.</a>
<img alt="Figure 3" class="xfigimg" src="/files/ftp_upload/55884/55884fig3.jpg" /> 
Figure 3: The templating tools prepared to ease the device fabrication: (a) Acrylic template: actual template (top) and its schematic cross-sectional view (bottom). (b) PDMS replica of the acrylic template: actual replica (top) and its schematic cross-sectional view (bottom). <a href="https://cloudflare-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/55884/55884fig3large.jpg" target="_blank">Please click here to view a larger version of this figure.</a>
<img alt="Figure 4" class="xfigimg" src="/files/ftp_upload/55884/55884fig4.jpg" /> 
Figure 4: Process flow for the fabrication of the patterned half of the device: (a) Place the PDMS template on a flat surface. Here, we used soda lime glass. (b) CaF2 is placed centered on top of the PDMS plug. (c-d) PDMS mold is placed face down towards the CaF2 with the fluidic chamber aligned to the center of the window. Ensure that all the elements are in good contact and aligned. (e-f) Casting of NOA via the in-let and allowing it to slowly fill the cavity. (g) NOA is cured by exposing it under UV light. Exposure dose may vary depending on the energy of the UV source used. (h) Carefully peel the PDMS mold and template to release the cured NOA. (i) Completed device layer with microfluidic structures in NOA. <a href="https://cloudflare-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/55884/55884fig4large.jpg" target="_blank">Please click here to view a larger version of this figure.</a>
<img alt="Figure 5" class="xfigimg" src="/files/ftp_upload/55884/55884fig5.jpg" /> 
Figure 5: Vacuum press setup for pressing the two halves of the device to form a complete device: (a) PDMS circular disc (8 mm diameter) and PDMS rectangle (4 x 2.5 cm) with pre-cut openings. Both are cut from a 1 mm thick PDMS sheet. (b) Overview of the layer-stack before pressing. (c) Overview of the vacuum press. <a href="https://cloudflare-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/55884/55884fig5large.jpg" target="_blank">Please click here to view a larger version of this figure.</a>
<img alt="Figure 6" class="xfigimg" src="/files/ftp_upload/55884/55884fig6.jpg" /> 
Figure 6: Mid-IR transmittance spectra of a bare CaF2 window (red), a patterned half of the device (blue), and a complete device (yellow). All three spectra exhibit excellent transparency to mid IR with transmittance larger than 80% <a href="https://cloudflare-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/55884/55884fig6large.jpg" target="_blank">Please click here to view a larger version of this figure.</a>
<img alt="Figure 7" class="xfigimg" src="/files/ftp_upload/55884/55884fig7.jpg" /> 
Figure 7: Patterned half of the device: (a) Example of a well formed microfluidic structure in NOA. Darker area on the window shows a well defined structure, showing clearly the central chamber, two reference chambers, and the channels. (b) Example of a poorly formed microfluidic structure in NOA due to undercuring. There is reflow of NOA as indicated by red arrows. One of the reference chambers is also missing (green arrow). <a href="https://cloudflare-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/55884/55884fig7large.jpg" target="_blank">Please click here to view a larger version of this figure.</a>
<img alt="Figure 8" class="xfigimg" src="/files/ftp_upload/55884/55884fig8.jpg" /> 
Figure 8: Microfluidic jig connecting the fabricated device to external fluidic circuitry. <a href="https://cloudflare-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/55884/55884fig8large.jpg" target="_blank">Please click here to view a larger version of this figure.</a>
<img alt="Figure 9" class="xfigimg" src="/files/ftp_upload/55884/55884fig9.jpg" /> 
Figure 9: Placement of PDMS mold during fabrication of the patterned half of the device: (a) Example of good placement of the PDMS mold on the CaF2 window. The window is in contact with only the protrusion on the PDMS mold, showing clearly the central chamber, two reference chambers, and the channels (indicated by the darker area). The diffraction gratings around the device and at the center of the central chamber are meant for a visual guide during placement of the PDMS mold. (b) Example of a bad placement of the PDMS mold on the CaF2 window. The darker area shows that there is also an unwanted area of the mold, which is in contact with the window, as indicated by the red arrows. <a href="https://cloudflare-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/55884/55884fig9large.jpg" target="_blank">Please click here to view a larger version of this figure.</a>

Watch this Scientific Journal Video about Soft Lithographic Procedure for Producing Plastic Microfluidic Devices with View-ports Transparent to Visible and Infrared Light at JoVE.com

Soft Lithographic Procedure for Producing Plastic Microfluidic Devices with View-ports Transparent to Visible and Infrared Light

A protocol for the fabrication of plastic microfluidic devices with transparent view-ports for visible and infrared light imaging is described.

Infrared (IR) spectro-microscopy of living biological samples is hampered by the absorption of water in the mid-IR range and by the lack of suitable ...

soft-lithographic-procedure-for-producing-plastic-microfluidic

Nanyang Technological University

Research

JoVE Journal

Bioengineering

13.0K Views.  National University of Singapore. A protocol for the fabrication of plastic microfluidic devices with transparent view-ports for visible and infrared light imaging is described.

Video: Soft Lithographic Procedure for Producing Plastic Microfluidic Devices with View-ports Transparent to Visible and Infrared Light

Microfluidic Devices for Characterizing Pore-scale Event Processes in Porous Media for Oil Recovery Applications

Microfluidic devices are versatile tools for studying transport processes at a microscopic scale. A demand exists for microfluidic devices that are resistant to low molecular-weight oil components, unlike traditional polydimethylsiloxane (PDMS) devices. Here, we demonstrate a facile method for making a device with this property, and we use the product of this protocol for examining the pore-scale mechanisms by which foam recovers crude oil. A pattern is first designed using computer-aided design (CAD) software and printed on a transparency with a high-resolution printer. This pattern is then transferred to a photoresist via a lithography procedure. PDMS is cast on the pattern, cured in an oven, and removed to obtain a mold. A thiol-ene crosslinking polymer, commonly used as an optical adhesive (OA), is then poured onto the mold and cured under UV light. The PDMS mold is peeled away from the optical adhesive cast. A glass substrate is then prepared, and the two halves of the device are bonded together. Optical adhesive-based devices are more robust than traditional PDMS microfluidic devices. The epoxy structure is resistant to swelling by many organic solvents, which opens new possibilities for experiments involving light organic liquids. Additionally, the surface wettability behavior of these devices is more stable than that of PDMS. The construction of optical adhesive microfluidic devices is simple, yet requires incrementally more effort than the making of PDMS-based devices. Also, though optical adhesive devices are stable in organic liquids, they may exhibit reduced bond-strength after a long time. Optical adhesive microfluidic devices can be made in geometries that act as 2-D micromodels for porous media. These devices are applied in the study of oil displacement to improve our understanding of the pore-scale mechanisms involved in enhanced oil recovery and aquifer remediation.

The goal of this procedure is to easily and rapidly produce a microfluidic device with customizable geometry and resistance to swelling by organic fluids for oil recovery studies. A polydimethylsiloxane mold is first generated, and then used to cast the epoxy-based device. A representative displacement study is reported.

Microfluidic devices are versatile tools for studying transport processes at a microscopic scale. A demand exists for microfluidic devices that are ...

Environment

A Microfluidic Platform to Study Bioclogging in Porous Media

Bacterial biofilms are found in several environmental and industrial porous media, including soils and filtration membranes. Biofilms grow under certain flow conditions and can clog pores, thereby redirecting the local fluid flow. The ability of biofilms to clog pores, the so-called bioclogging, can have a tremendous effect on the local permeability of the porous medium, creating a pressure buildup in the system, and impacting the mass flow through it. To understand the interplay between biofilm growth and fluid flow under different physical conditions (e.g., at different flow velocities and pore sizes), in the present study, a microfluidic platform is developed to visualize biofilm development using a microscope under externally-imposed, controlled physical conditions. The biofilm-induced pressure buildup in the porous medium can be measured simultaneously using pressure sensors and, later, correlated with the surface coverage of the biofilm. The presented platform provides a baseline for a systematic approach to investigate bioclogging caused by biofilms in porous media under flow conditions and can be adapted to studying environmental isolates or multispecies biofilms.

The present protocol describes a microfluidic platform to study biofilm development in quasi-2D porous media by combining high-resolution microscopy imaging with simultaneous pressure difference measurements. The platform quantifies the influence of pore size and fluid flow rates in porous media on bioclogging.

Bacterial biofilms are found in several environmental and industrial porous media, including soils and filtration membranes. Biofilms grow under certain ...

Fabrication of Refractive-index-matched Devices for Biomedical Microfluidics

The use of microfluidic devices has emerged as a defining tool for biomedical applications. When combined with modern microscopy techniques, these devices can be implemented as part of a robust platform capable of making simultaneous complementary measurements. The primary challenge created by the combination of these two techniques is the mismatch in refractive index between the materials traditionally used to make microfluidic devices and the aqueous solutions typically used in biomedicine. This mismatch can create optical artifacts near the channel or device edges. One solution is to reduce the refractive index of the material used to fabricate the device by using a fluorinated polymer such as MY133-V2000 whose refractive index is similar to that of water (n = 1.33). Here, the construction of a microfluidic device made out of MY133-V2000 using soft lithography techniques is demonstrated, using O2 plasma in conjunction with an acrylic holder to increase the adhesion between the MY133-V2000 fabricated device and the polydimethylsiloxane (PDMS) substrate. The device is then tested by incubating it filled with cell culture media for 24 h to demonstrate the ability of the device to maintain cell culture conditions during the course of a typical imaging experiment. Finally, quantitative phase microscopy (QPM) is used to measure the distribution of mass within the live adherent cells in the microchannel. This way, the increased precision, enabled by fabricating the device from a low index of refraction polymer such as MY133-V2000 in lieu of traditional soft lithography materials such as PDMS, is demonstrated. Overall, this approach for fabricating microfluidic devices can be readily integrated into existing soft lithography workflows in order to reduce optical artifacts and increase measurement precision.

This protocol describes the fabrication of microfluidic devices from MY133-V2000 to eliminate artifacts that often arise in microchannels due to the mismatching refractive indices between microchannel structures and an aqueous solution. This protocol uses an acrylic holder to compress the encapsulated device, improving adhesion both chemically and mechanically.

The use of microfluidic devices has emerged as a defining tool for biomedical applications. When combined with modern microscopy techniques, these devices ...

Engineering

A Microfluidic Platform for High-throughput Single-cell Isolation and Culture

Studying the heterogeneity of single cells is crucial for many biological questions, but is technically difficult. Thus, there is a need for a simple, yet high-throughput, method to perform single-cell culture experiments. Here, we report a microfluidic chip-based strategy for high-efficiency single-cell isolation (~77%) and demonstrate its capability of performing long-term single-cell culture (up to 7 d) and cellular heterogeneity analysis using clonogenic assay. These applications were demonstrated with KT98 mouse neural stem cells, and A549 and MDA-MB-435 human cancer cells. High single-cell isolation efficiency and long-term culture capability are achieved by using different sizes of microwells on the top and bottom of the microfluidic channel. The small microwell array is designed for precisely isolating single-cells, and the large microwell array is used for single-cell clonal culture in the microfluidic chip. This microfluidic platform constitutes an attractive approach for single-cell culture applications, due to its flexibility of adjustable cell culture spaces for different culture strategies, without decreasing isolation efficiency.

Here, we present a protocol for isolating and culturing single cells with a microfluidic platform, which utilizes a new microwell design concept to allow for high-efficiency single cell isolation and long-term clonal culture.

Studying the heterogeneity of single cells is crucial for many biological questions, but is technically difficult. Thus, there is a need for a simple, yet ...

Microfluidic Fabrication Techniques for High-Pressure Testing of Microscale Supercritical CO2 Foam Transport in Fractured Unconventional Reservoirs

Pressure limitations of many microfluidic platforms have been a significant challenge in microfluidic experimental studies of fractured media. As a result, these platforms have not been fully exploited for direct observation of high-pressure transport in fractures. This work introduces microfluidic platforms that enable direct observation of multiphase flow in devices featuring surrogate permeable media and fractured systems. Such platforms provide a pathway to address important and timely questions such as those related to CO2 capture, utilization and storage. This work provides a detailed description of the fabrication techniques and an experimental setup that may serve to analyze the behavior of supercritical CO2 (scCO2) foam, its structure and stability. Such studies provide important insights regarding enhanced oil recovery processes and the role of hydraulic fractures in resource recovery from unconventional reservoirs. This work presents a comparative study of microfluidic devices developed using two different techniques: photolithography/wet-etching/thermal-bonding versus Selective Laser-induced Etching. Both techniques result in devices that are chemically and physically resistant and tolerant of high pressure and temperature conditions that correspond to subsurface systems of interest. Both techniques provide pathways to high-precision etched microchannels and capable lab-on-chip devices. Photolithography/wet-etching, however, enables fabrication of complex channel networks with complex geometries, which would be a challenging task for laser etching techniques. This work summarizes a step-by-step photolithography, wet-etching and glass thermal-bonding protocol and, presents representative observations of foam transport with relevance to oil recovery from unconventional tight and shale formations. Finally, this work describes the use of a high-resolution monochromatic sensor to observe scCO2 foam behavior where the entirety of the permeable medium is observed simultaneously while preserving the resolution needed to resolve features as small as 10 &#181;m.

Microfluidic Fabrication Techniques for High-Pressure Testing of Microscale Supercritical CO2 Foam Transport in Fractured Unconventional Reservoirs

This paper describes a protocol along with a comparative study of two microfluidic fabrication techniques, namely photolithography/wet-etching/thermal-bonding and Selective Laser-induced Etching (SLE), that are suitable for high-pressure conditions. These techniques constitute enabling platforms for direct observation of fluid flow in surrogate permeable media and fractured systems under reservoir conditions.

Pressure limitations of many microfluidic platforms have been a significant challenge in microfluidic experimental studies of fractured media. As a ...

Window on a Microworld: Simple Microfluidic Systems for Studying Microbial Transport in Porous Media

Microbial growth and transport in porous media have important implications for the quality of groundwater and surface water, the recycling of nutrients in the environment, as well as directly for the transmission of pathogens to drinking water supplies. Natural porous media is composed of an intricate physical topology, varied surface chemistries, dynamic gradients of nutrients and electron acceptors, and a patchy distribution of microbes. These features vary substantially over a length scale of microns, making the results of macro-scale investigations of microbial transport difficult to interpret, and the validation of mechanistic models challenging. Here we demonstrate how simple microfluidic devices can be used to visualize microbial interactions with micro-structured habitats, to identify key processes influencing the observed phenomena, and to systematically validate predictive models. Simple, easy-to-use flow cells were constructed out of the transparent, biocompatible and oxygen-permeable material poly(dimethyl siloxane). Standard methods of photolithography were used to make micro-structured masters, and replica molding was used to cast micro-structured flow cells from the masters. The physical design of the flow cell chamber is adaptable to the experimental requirements: microchannels can vary from simple linear connections to complex topologies with feature sizes as small as 2 &mu;m. Our modular EcoChip flow cell array features dozens of identical chambers and flow control by a gravity-driven flow module. We demonstrate that through use of EcoChip devices, physical structures and pressure heads can be held constant or varied systematically while the influence of surface chemistry, fluid properties, or the characteristics of the microbial population is investigated. Through transport experiments using a non-pathogenic, green fluorescent protein-expressing Vibrio bacterial strain, we illustrate the importance of habitat structure, flow conditions, and inoculums size on fundamental transport phenomena, and with real-time particle-scale observations, demonstrate that microfluidics offer a compelling view of a hidden world.

Microfluidic devices can be used to visualize complex natural processes in real time and at the appropriate physical scales. We have developed a simple microfluidic device that mimics key features of natural porous media for studying growth and transport of bacteria in the subsurface.

Microbial growth and transport in porous media have important implications for the quality of groundwater and surface water, the recycling of nutrients in ...

Biology

Multi-step Variable Height Photolithography for Valved Multilayer Microfluidic Devices

Microfluidic systems have enabled powerful new approaches to high-throughput biochemical and biological analysis. However, there remains a barrier to entry for non-specialists who would benefit greatly from the ability to develop their own microfluidic devices to address research questions. Particularly lacking has been the open dissemination of protocols related to photolithography, a key step in the development of a replica mold for the manufacture of polydimethylsiloxane (PDMS) devices. While the fabrication of single height silicon masters has been explored extensively in literature, fabrication steps for more complicated photolithography features necessary for many interesting device functionalities (such as feature rounding to make valve structures, multi-height single-mold patterning, or high aspect ratio definition) are often not explicitly outlined. 
Here, we provide a complete protocol for making multilayer microfluidic devices with valves and complex multi-height geometries, tunable for any application. These fabrication procedures are presented in the context of a microfluidic hydrogel bead synthesizer and demonstrate the production of droplets containing polyethylene glycol (PEG diacrylate) and a photoinitiator that can be polymerized into solid beads. This protocol and accompanying discussion provide a foundation of design principles and fabrication methods that enables development of a wide variety of microfluidic devices. The details included here should allow non-specialists to design and fabricate novel devices, thereby bringing a host of recently developed technologies to their most exciting applications in biological laboratories.

Multilayer microfluidic devices often involve the fabrication of master molds with complex geometries for functionality. This article presents a complete protocol for multi-step photolithography with valves and variable height features tunable to any application. As a demonstration, we fabricate a microfluidic droplet generator capable of producing hydrogel beads.

Microfluidic systems have enabled powerful new approaches to high-throughput biochemical and biological analysis. However, there remains a barrier to ...

Rapid Fabrication of Custom Microfluidic Devices for Research and Educational Applications

Microfluidic devices allow for the manipulation of fluids, particles, cells, micro-sized organs or organisms in channels ranging from the nano to submillimeter scales. A rapid increase in the use of this technology in the biological sciences has prompted a need for methods that are accessible to a wide range of research groups. Current fabrication standards, such as PDMS bonding, require expensive and time consuming lithographic and bonding techniques. A viable alternative is the use of equipment and materials that are easily affordable, require minimal expertise and allow for the rapid iteration of designs. In this work we describe a protocol for designing and producing PET-laminates (PETLs), microfluidic devices that are inexpensive, easy to fabricate, and consume significantly less time to generate than other approaches to microfluidics technology. They consist of thermally bonded film sheets, in which channels and other features are defined using a craft cutter. PETLs solve field-specific technical challenges while dramatically reducing obstacles to adoption. This approach facilitates the accessibility of microfluidics devices in both research and educational settings, providing a reliable platform for new methods of inquiry.

Here we present a protocol to design and fabricate custom microfluidic devices with minimal financial and time investment. The aim is to facilitate the adoption of microfluidic technologies in biomedical research laboratories and educational settings.

Microfluidic devices allow for the manipulation of fluids, particles, cells, micro-sized organs or organisms in channels ranging from the nano to ...

Microfabrication of Implantable Optics Integrated in a Microstructured Imaging Window for Advanced In Vivo Imaging

In the context of biomaterials and drug testing in animal models, this study presents a streamlined protocol for fabricating a novel implantable integrated imaging window. The micro-device comprises a sophisticated system of microlenses coupled with micro-scaffolds specifically designed for in vivo quantification of the immune response using advanced non-linear excitation microscopy. The protocol is based on two-photon polymerization (2PP) of the biocompatible photoresist SZ2080, which enables the fabrication of micro-scaffolds and micro-lenses in a continuous sequence to enhance manufacturing efficiency and precision. To further improve speed, accuracy, and structural integrity, a hybrid optics fabrication approach was implemented, involving the 2PP of the microlens outer shell followed by UV bulk crosslinking of the inner core. This innovative technique optimizes the optical properties of the microlenses while streamlining the production process. The resulting micro-device demonstrates high reproducibility and mechanical stability, making it an effective method for prototyping microscale optical systems for a range of biomedical applications.

This protocol describes the fabrication of an implantable integrated imaging window using 3D laser printing. The window consists of a system of microlenses coupled with micro-scaffolds. The method involves two-photon polymerization (2PP) of the biocompatible photoresist SZ2080 in a continuous sequence, optimizing manufacturing efficiency and alignment between the different components.

In the context of biomaterials and drug testing in animal models, this study presents a streamlined protocol for fabricating a novel implantable ...

Microfabrication via Photolithography

The fabrication of BioMEM&#160;devices is often done using a microfabrication technique called photolithography. This widely used method utilizes light to transfer a pattern onto a silicon wafer, and provides the basis for the fabrication of many types of BioMEM&#160;devices.
This video presents the photolithography technique, shows how the process is performed in the cleanroom, and introduces some applications of the process.

Microfabrication of BioMEM Devices via Photolithography

The fabrication of BioMEM&#160;devices is often done using a microfabrication technique called photolithography. This widely used method utilizes light to ...