How to make glass microfluidics device? chemical wet etching and room temperature bonding

[First part of this post is here where I wrote step by step procedure for mask design and photolithography.]

After completing the photolithographic procedure, we use two different solutions for selectively removing chromium and glass from the channel network on the bottom substrate. 

Chemical wet etching solutions. On left is BOE with temperature control
and on right is Chrom etchant. Middle one is DI water.

At first, chromium layer from channel network is removed by immersing the substrate plate into a Teflon coated jar containing chromium etchant solution for about 15 min. One should note that leaving substrate plate in this solution for long time (hours and hours-this happens when you forget to remove it) could eat away even the photoresist layer from the entire plate. We buy the chrome etchant from Transene Company, INC, MA, USA and according to manufacturer specification the solution contains ceric sulfate (5-10%), nitric acid (5-10%), sulfuric Acid (1-5%), water (~ 75%). The entire substrate is then washed with DI water and dried by blowing N₂ gas. At this point one can clearly see through the substrate in channel patterning region.

Cartoon depicting the removal of photoresist and chromium layer from channel pattern

After this step, we cut the substrate into individual chips (1" x 2") using a glass-cutter (altogether 8 chips from one 8” x 8” substrate). Then, put extra layer of photoresist, dry in oven (80 ^o C), take out from the oven, and let it cool.

The substrate plate-individual chip is then immersed into a 1:10 buffered oxide etchant (BOE) solution bought from Transene Company, INC, MA, USA. According to the company specifications, this solution contains a mixture of hydrofluoric (HF) (1-25%), ammonium fluoride (NH₄F) (2-40%), distilled water. Hydrofluoric acid is a hazardous chemical, therefore must be handled with extra precaution. The BOE solution is continuously stirred at the rate of 120 rpm using a magnetic stirrer and the temperature is set at 55 ^oC. The substrate plate is removed from the BOE solution at a certain interval of time ~10-15 min (depending on the etching rate of BOE and channel depth desire), washed with DI water, and is dried with N₂. With our settings, etching rate is ~0.5 micron per minute. 

However, this rate can be manipulated based on your need. 

Cartoon showing the substrate plate after removing glass material.
Remember the glass etching is in all direction (not like in this cartoon)

The depth of the channel is measured using a XP series stylus profiler (Ambios Technology, CA, USA). This step is repeated until one gets desired channel depth.  

Profiler

Once desired channel depth is obtained, ~1 mm diameter access holes were punched at the channel terminals using a microabrasive power blasting system (Vaniman) by blowing sand particles from back side of the substrate. The miroabrasive system utilizes mechanical erosion on the substrate by bombarding with high-kinetic energy sand particles and results in conically shaped holes. This step is followed by removing the photoresist and chromium layer on remaining parts of the bottom substrate by using acetone and chromium etchant, respectively. 

Sandblaster 

Bonding two glass plates: The etched channels in the bottom substrate were sealed using cover plate. The two plates (bottom substrate and cover plate) were cleaned and brought together in a beaker containing DI water for bonding. They were removed from the beaker together with a tweezer, water was removed by gently pressing the plates with hand in paper towel, and then put under couple of heavy books for ~2 hrs. Then the bonded chip was kept in an oven at 80 ^oC for ~1hrs to strengthen the bonding between two glass plates. Bonding is a crucial step to create closed fluidic microchip networks. 

The bonding strength between two surfaces is proportional to the density of individual chemical bonds established between the two plates. Our bonding method is simple and does not require clean room facilities, programmed high-temperature furnaces, pressurized water sources, and adhesives. The bonding process can be completed in ~3 hrs. One of the most important factors affecting successful bonding of planner glass chips is the cleanliness of the bonding surfaces of glass substrates. Unsuccessful bonding events are often associated with solid particles or organic material remaining on the glass surfaces, for example, dust, residual photoresist, chromium or glass particles etc., prior to bonding. Some other critical factors for glass bonding are flatness of the glass plates and chip area.

no-instrument measurement in paper-based device

Paper-based devices (PBDs) have emerged as a new class of inexpensive devices with potential use in medical diagnostics, environmental applications etc. They are not only inexpensive but also proven to be sensitive, quantitative, and reliable in many applications.

The colorimetric detection is one of the popular methods of quantitation of analytes in PBDs. Color developing reagents are used on the micro-reaction chambers on paper. The color developing reagent then reacts with the analyte of interest when the later is applied specifically and quantitatively. Simple and widely available instrumentations like a office scanner or a camera (could be a mobile phone camera) are used to capture the color which is then quantitated by measuring the signal using image processing software.  If you don't have camera and/ or image processing software, don't worry. Scientists from Colorado State University have developed even a simpler method to quantitate analytes. This interesting work has recently been published in Lab on a Chip Journal.

The new technique relies on the length of the color developed. They call this as a simple distance-based detection. You just need a ruler to quantitate the analyte concentration: no camera, no scanner, no software.

How does this work?

1.

First of all create a paper-based device. In their work, they have used wax printing to make PBDs. Their design has a straight channel, like a thermometer. There is one circular reservoir at one end. This reservoir is used for sample addition and/or transfer of analyte to the detection zone (long conduit).

2.



Colorimetric detection reagents are deposited along the flow channel. They tested two different methods for this purpose: spray application or pipetting. Once the reagents are dry, the device is ready to use.

3.

Sample solution in water is then added to the sample reservoir which flows into the detection/flow channel. The analyte reacts with reagent and develops color. Once all of the analyte is consumed, the color development stops. Analyte quantification is then done by measuring the length of the colored region in the detection zone by a ruler.

Authors of this paper have demonstrated their technique by measuring three different analytes: nickel, reduced glutathion, and glucose.

This new measurement technique is claimed to be accurate and sensitive compared to conventional methods.

In this figure, you can see that higher the concentration of analyte-the longer is the color developed region in detection zone.