Intelligent Transportation of Heterogeneous droplets in

DMFB: A Preferential Deviation Based Technique

Pranab Roy Sarit Chakrabarty Rupam Bhattacharya
J.K.Lakshmipat University Govt. College of Engineering Institute of Engineering and Management
Jaipur,India and Leather Technology, Kolkata, India Kolkata,India
pranab.roy@jklu.edu.in sarit@gcelt.gov.in th_rup@yahoo.co.in

Priyatosh Jana Paramita Dey Sushanta Chakraborty

Haldia Institute of Technology Govt. College of Engineering IIEST,Shibpur, India
West Bengal,India and Ceramic Technology, Kolkata,India sc@cs.iiests.ac.in
priyatoshjana@gmail.com dey.paramita77@gmail.com

Abstract— In order to enhance the performance of DMFBs, the a unit cell in the digital microfluidic biochip. This set of electrodes
parallel routing of droplets plays a crucial role. By implementing forms two parallel plates. The base plate, that is a ground plate built
this approach in a single bio-assay, we can effectively optimize the with a set of separately controllable electrode whereas upper plate is
2023 International Symposium on Devices, Circuits and Systems (ISDCS) | 979-8-3503-1504-2/23/$31.00 ©2023 IEEE | DOI: 10.1109/ISDCS58735.2023.10153566

latest arrival time. However, the various fluidic constraints present glazed by continuous ground electrode. As shown in Figure 1, a
on DMFB platform raise different challenges such as collisions and droplet placed on a hydrophobic surface of an electrode. These
contaminations, especially when dealing with heterogeneous droplet droplets are transmitted by incorporating a control voltage to a
routing. Therefore, it becomes imperative to devise an efficient neighboring electrode as well as deactivating the electrode on which
algorithm that not only satisfies all the constraints but also droplet is placed. This method produces interfacial tension gradients
minimizes issues like stalling, collisions, and contaminations caused which transmit the droplet towards charged electrode. In this way,
by parallel routing. By tackling these obstacles head-on, we can by applying electrowetting process, the droplets can be routed to any
significantly improve the overall efficiency and reliability of position of the two-dimensional array of electrode.
DMFBs. In this work, we propose an algorithm for heterogeneous Digital microfluidics offers several advantages over continuous
droplet transportation. The proposed intelligent deviation preference flow based digital microfluidics. In digital microfluidic, each
scheme serves as a valuable tool in enhancing the overall independent droplets are under operation in a discrete manner
performance and reliability of the system, allowing for seamless instead of a flow of the entire fluids, and thus we can have more
parallel routing and improved cell utilization with latest arrival time. precise control on those droplets individually.
Requirement of sample and reagent reduced to nano liter to picolitre
Keywords—Digital Microfluidic Biochip, Droplet volume, which in turn reduces the operational cost and almost make
Transportation, Cross contamination, Deviation preference. digital microfluidic based biochip a disposable one.
A group of microfluidic operations such as mixing of droplets,
I. INTRODUCTION splitting, routing or transportation of droplets to any location within
two dimensions can be easily executed in digital microfluidic
Microfluidics is a field of science dedicated to the manipulation and
biochips.
control of fluids within networks of channels, typically ranging from
Such devices are highly reconfigurable and can perform several
microliters (10-6) to picoliters (10-12), with dimensions in the range of
biomedical and pathological tests and operations in parallel.
tens to hundreds of micrometers. The utilization of microfluidic
devices in biomedical research and the development of clinically
relevant technologies offers numerous advantages. These devices
significantly reduce reagent consumption, provide precise control
over mixing and sample manipulation, enable the integration and
parallel automation of multiple assays, and facilitate high-precision
detection through imaging and tracking. Lab-on-chips, also known as
microfluidic biochips, have emerged as a cutting-edge replacement
for traditional laboratory processes in various biomedical
applications. However, in terms of adaptability, reconfigurability,
and fault tolerance the prior generation of continuous flow-based
biochips fall short with various limitations. Digital microfluidic
biochips (DMFBs), the latest type of biochips, have been designed
to tackle such issues.

Unlike their predecessors, DMFBs utilize liquid droplets instead of

continuous liquid flow, offering enhanced flexibility, broader
reconfigurability, and improved fault tolerance. Digital microfluidics
Figure 1. Unit cell of EWOD-based digital microfluidic biochip
is sometimes called planar microfluidics because it involves
movement, merging, and dividing droplets on a planar 2-
Execution of bioassays in parallel on a digital-microfluidic
diamensional surface. There are two means of actuation: acoustic
biochip needs system-level design automation tools to handle the
and electric. In digital microfluidics, the means of actuation
biochip’s increasing design complexity. The major microfluidic
followed are largely electric. More precisely, this is electrowetting
operations namely mixing, merging, splitting, transportation, and
on dielectrics. The concept of electrowetting is deployed to control
detection are accomplished within the 2D layout of the DMFB. Out
and manipulates that micro or nanoliter volume droplets on a 2D
of these operations droplet routing or transportation is considered as
array of electrode [8]. This unit volume of droplets mainly consists
one of the major microfluidic operations.
of biochemical or biomedical samples. A couple of electrodes form

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 For the biological assays to produce appropriate results, seamless
transportation of the discrete droplets is the most crucial criteria. In
the DMFB architecture, a droplet migrates from the source electrode
to the target electrode over the pre-calculated routes with the help of
electrode actuations. The characteristics of the droplets may be
heterogeneous while they are being transported in parallel using 2
or 3 pin nets. The term heterogeneous denotes here that each net is
either constructed for the parallel execution of multiple bioassays or
is derived from various participants for the execution of a single
bioassay schedule.
II. MOTIVATION AND GOALS
During the transportation of droplets, there is a possibility of
unforeseen mixing between droplets when their route paths
intersect. This phenomenon is referred to as cross-contamination.
When functional droplets drops residual leftover on the electrode
cells as they pass through, and if other droplets incidentally passes
through any such cells along their path, the phenomenon of cross-
contamination takes place. Therefore, when multiple droplets are
routed in parallel, it is a significant challenge to ensure that they
arrive at the desired location while intelligently avoiding
contamination.
To avoid cross-contamination, there are two approaches that can be
taken. The first is to prevent contamination entirely during droplet
path estimation. The second is to handle overall contamination by
incorporating wash droplet routing. In either case, the route paths of
different droplets should ideally be kept separate from each other to
avoid cross-contamination.
In transportation of droplets heterogeneous in nature, the primary
focus is on determining disjoint transportation paths to prevent
undesired contamination due to crossovers of droplets. Such
contamination can lead to inaccurate detection followed by analysis
of the output, resulting in significant errors in the diagnosis process.
Therefore, it is necessary to develop intelligent techniques that may
allow for the estimation of route paths that are disjoint from each
other, minimizing or eliminating contamination.
The main objectives for this work are as follows:
• Estimating disjoint route paths for each droplet to avoid
cross-contamination.
• Allowing minimal crossover if disjoint paths are not
feasible.
• Developing intelligent deviation techniques to avoid
crossovers while minimizing the deviation to ensure
optimal utilization of cells.
• Maintaining optimal routing parameters, such as average
arrival time and latest arrival time, while avoiding
contamination.
The rest of the paper is organized as follows. Section III discusses
contemporary works in cross-contamination of biochips. Section IV
states the proposed method and Section V depicts an example.
Section VI shows the experimental results followed by brief
analysis. Finally, section VII concludes the work with suggestions
for future improvements.
III. CONTEMPORARY CONTRIBUTIONS
Droplet transportation in digital microfluidic biochips (DMFBs)
poses significant challenges related to route time, collisions, and
cross-contamination. Among these challenges, cross-contamination
is particularly problematic for heterogeneous droplets as it directly
affects the accuracy of detection outcomes.
In a study by researchers [9], an algorithm was developed to address
efficient routing of heterogeneous droplets in DMFBs with a focus
on contamination elimination. The algorithm begins by considering
two Manhattan paths and selects the path with the least
contamination. It then allows detours to further minimize
contamination. Experimental results demonstrated a notable
improvement in contamination avoidance. Additionally, the
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algorithm aims to satisfy timing constraints and a complete routing
simulator was implemented using Java.
Another approach presented in [10] aims to detect and minimize
routing errors. They propose a heuristic to reduce overall time for
routing and the number of utilization of electrodes in the
transportation process, as described in [11]. The proposed algorithm
was tested using standard benchmarks, showing promising
outcomes.
To eliminate or minimize the probability of errors or defects
resulting from multiple redundant resource usage, a routing scheme
was formulated in [12]. This scheme enables concurrent routing of
multiple droplets while avoiding redundant resource utilization. A
variable is assigned to each electrode, and its value is modified after
each use in each iteration. Experimental simulations demonstrated
significant improvements compared to existing approaches.
In [13], a technique for routing of test droplets was proposed. The
technique activates specific control pins of electrodes based on
allocation methods, allowing droplets to move within the 2D layout
of DMFB. Additionally, an algorithm for multi-objective
optimization was developed in [14] to reduce the use of multiple
resources during bioassay execution in a DMFB. Furthermore, [15]
introduces a heuristic-based method for parallel routing of droplets
and electrode actuation. This method minimizes the number of
electrodes used and achieves the required completion time for
routing.
A technique for routing of parallel droplets, combined with a method
for cross-contamination avoidance, was presented in [16]. For
unavoidable cross-contaminations, the authors proposed routing of
wash droplets through contaminated sites. The method utilizes a
heuristics based on hierarchical partitioning, creating unbalanced
tree structure using routing nets at different nodes.
In [17], a contamination-aware droplet routing method is described,
utilizing a k-shortest path routing approach to minimize
contamination within a sub-problem. Number of wash droplets are
thereby deployed to remove native contaminations in cells. The
method also employs a look-ahead prediction technique to identify
contaminations between droplets from different sub-problems.
For simultaneous path allocation to a set of droplets, a routing
algorithm was developed in [18]. The algorithm incorporates stall to
avoid collisions between droplets routed in parallel, accommodating
both two-pin and three-pin nets.
Addressing the issue of cross-contamination, [19] proposes a
solution by employing k-shortest path techniques for routing to
minimize complexities and utilization of cells, thereby causing
reduction in the number of spots marked as contaminated.
In [20], a technique is introduced along with genetic operators for
the NSGAII for handling the issue of droplet routing. The objective
is to optimize the number of used cells and the last droplet arrival
time. Furthermore, [21] presents an exact routing method that uses
the observation that blockages usually occur at specific intervals.
This approach guarantees exact solutions by determining routing
schemes with the fewest time steps.
In this paper we proposed a route zone-based deviation preference
technique to access suitable deviation of routes for intelligent
crossover avoidance. The method shows improvements over the
contemporary techniques for cross contamination aware droplet
transportation.
IV. PROPOSED ROUTE ZONE BASED PATH ESTIMATION
For a specific DMFB 2 dimensional layout - locations of source,
mixer and target for a net already are predtermined. Hence the
route zone of a 2 pin net is described as a rectangle formed
comprising of the cells with respective source location and target
location being its opposite end points. diagonally
Initially Manhattan paths for each is planned. Next, estimated are
the requisite deviations to avoid modules termed as hard blockages.
Finally, we identify the tentative crossover sites – where multiple
 net paths share the same cell locations. For these conditions, certain
net routes involved in the crossover are deviated eventually.
Out of multiple nets it is found to be necessary to determine
preference for deviation with a number of nets. Such nets- once
deviated can formulate disjoint paths – to reduce or eliminate net
crossovers. For any nets - deviation preference may be determined
following a route zone based technique.
The technique proposes to plot one route zone per single 2 pin
net. Thereby estimations are done for route zones’ intersections.
Such intersection involving the number of cell coverage may display
an initial estimation for cross contamination possibilities. So, such
intersection can allow only a single of the multiple intersecting nets
to pass along. Deviation is necessary for the remaining ones. Here
attempt is made to identify the nets having minimum area of
intersection within its own route zone. If a net involves in higher
number of intersections within its stipulated route zone – it turns out
to be a suitable candidate for deviation in its route path.
Fig 2.a) Nets having preference in deviation;
Fig 2.b) Net transportation routes having no cross-contamination.
Formulation of Problem
We consider a square DMFB having 2D layout of n x n dimension.
pre-located 2 pin nets are = x and pre-located 3 pin nets are =y.
We resolve them into a set of 2 pin nets m = x + 3y.
Now m routing zones are formed (one for each 2 pin net).
Our goal is to estimate m route paths having optimal deviation.
These paths might result in minimal number of cross-
contaminations combined with possible shortest length path for
every net involved.
Path Estimation Process
Given number of Two pin nets = m.
For every single net a Route Zone is formed.
number of route zones formed = m.
However, intersection of route zones 3 pin nets are not considered.
1. We assume intersection regions within these route zones are = s.
route zone graph named as GRZ (D, E) is drawn.
each vertex d ε D represents one Route zone and zone intersection
is represented by each edge e ε E.
2. We define GRZ has m number of vertices and s number of edges.
3. Each edge e is assigned with weight w. Weight w is computed as
= number of cells involved in intersection region represented by e.
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4. For each net i. Manhattan path from respective 2 pin nets is
plotted.
5. For specific paths running through hard blockages - deviations
using Hadlock’s technique is initiated to reach the targeted
destination
6. This tentatively ensures possible shortest paths to the target with
least number of diversions.
7. Respective route paths length for each net is computed.
8. Based on the graph GRZ - a table is formed listing of each node
of the graph. In rest of the columns, respective degree for each node,
path length computed so far and the overall weight for respective
edges associated to the total intersection for any single net is
enlisted.
9. Identified are the nets having maximum degree. Maximum degree
of nets are assigned with higher order of preference for deviation.
With two or more nets having same degree – higher preference is
assigned to the one with lower number of cells of intersection
coverage depicted as total weight for the edge under consideration.
10. Nets with equal degree and equal overall edge weight – higher
preference for deviation goes to one covering lower route distance.
11. Based on the assigned preferences, the path for net is plotted
with highest preference for deviation together with minimum
possible deviation.
12. Check for contaminations whether being reduced.
13. Continue deviation for nets with next level of preference.
14. Process of path estimation is terminated when no crossovers are
left or possible deviation for all the route paths for each net is
completed. (Sequentially as per the deviation preference)
15. If still there exists any further contamination – we consider it as
native contamination. These has been handled using wash droplets
with intelligent schedules planned afterwards.
Route Implementation
Once the path estimation phase is completed – the estimated route
information within the 2D layout is stored. Accordingly the route
implementation or detailed routing phase starts.
Step1 - Sort all the nets in descending order of their estimated
distance.
Step 2 – Identify all friction regions for the estimated route paths
Step 3 – For each friction region
Identify the route segments of the two involved in
friction with lower path length
Step 4 – Plan deviation for the identified route path
Step 5 – If deviation is hindered by another path segment of the
third net
Perform deadlock detour (refer to algorithm 3.1.i)
Step 6 – If deviation results in additional contaminations.
Plan deviation for the route segment of the net with
higher path length
Step 7 – If this involves contaminations.
check number of contaminations generated for
deviation of both segments
Step 8 – Accept deviation for the segment with lower number
of contamination
Step 9 – If no of contaminations are same in both cases.
Accept deviation for the segment with lower path
length
Step 10 – Accept such contamination as native contamination
(unavoidable)
Step 11 – Assume timestamp t=0 for each net.
Step 12 – Sort all nets in descending order of finally estimated path
after generated deviations.
Step 13 – Assign preference in ascending order to each net in sorted
sequence
Step 14 – Start with net number 1 in the sorted sequence
Step 15 – Perform route along the estimated with each increment
of timestamp by 1 for each transition between two adjacent cells.
 Step 16 – If collision is observed or fluidic constraints violation
occurs
perform stall of the net with lower preference
Step 17 – Continue transportation with increment of timestamp
until target is reached
Step 18 – Compute arrival time for the specific net
Step 19 – Continue for the next net number in the sorted sequence
Step 20 – Repeat from step 14 – 19 until transportation of all nets
are complete.
Step 21 – Identify the latest arrival time (e) (f)
Step 22 – If latest arrival time violates timing constraint
Perform recheck of the route path with alteration of
necessary deviation or stall
Step 23 – If violation persists
Identify the nets causing deviation and re-estimate
their route paths
Step 24 – If timing constraint violation still persists
back annotate for change in placement
procedure within the 2D layout
Else (g) (h)
Compute average arrival time
Compute cell utilization
Compute remaining contamination in
the overall transportation process.

V. AN ILLUSTRATIVE EXAMPLE

A 16 x 16 layout of a DMFB 2D layout is considered here. It contains

preplaced source and target locations for 6 droplets- a, b, c, d, e, f
each referring to one 2 pin net as shown in figure 3 a).Four
microfluidic modules are placed within the layout viz. M1, M2, M3,
M4 . Hence Formed are the 6 routing zones, for the specified nets
(A,B,C,D,E,F) [ figure 3 b)]. (i)

(a) (b)

(j)

Fig. 3 a) preplaced 2D Layout of DMFB. b ) Each net route zone

within the Layout. c) Intersection areas among route zones d)
Plotted intersection graph for the route zones e) Initial plan of routes
with contaminations. f) route path intersection graph g) After
deviation Route path being re-estimated. h) Additional deviation of
net a resulted in increase in overall path length . i) Estimated path
(c) (d) after iterative movement with preferential deviation. j) The
implementation of final route with timestamps showing the
computed arrival time and requisite stalls

Eventually intersection among route zones and the shaded

intersection regions are represented in figure 3 c). These are named
as as A-B – implying intersection of zones A and B and with others
so on. Assigned are the route deviation preferences according to the
technique proposed as stated in section IV. We form the intersection
graph for Route zone GRZ (V, E). In this graph - each vertex v

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 presents the route zone and corresponding intersection is contaminations, overall Arrival time and utilization of Cells for the
represented by each edge e. Assigned are the weight w to respective 4 testbenches with two of the state of the art contemporary works.
edges. The weight w presents the number of cells involved in the We observe marginal deterioration in the contamination number
zone of intersection. For zones having equal degrees, one with lower with the proposed method. This is due to the fact that we never
cell coverage is assigned with higher deviation preference. The compromised upon the other route parameters just in order to avoid
reason is that - scope for finding alternate cells for contamination crossovers – with application of deviation preference techniques
avoidance within the intersection zone is reduced. This is more stated above.
advantageous specifically for the intersection regions comprising of
total cells lower in number.
Manhattan path estimated with deviation is plotted only to Table 2. result of simulation for transportation of droplets using
divert from the hard blockages /modules. The initial routes are benchmark suite III
shown in figure 3 e) with possible contaminations (while they
remain within Route zones- deviation is being considered only to
bypass hard blockages).
The Graph for path intersection GInt (V, E) is displayed in figure
3 f). Here vertex v presents a 2 pin net, while each edge e is assigned
for intersection between paths. A box relates to each net presents
estimated Manhattan distance for each respective path of each net
so far being estimated.
Now We arrive at the final phase of bypassing the contamination.
Hence route Zone A is found to be the most preferred deviation Table 3. route simulation results comparison with contemporary
candidate. Next deviation preference goes to zone F. The respective works
result is presented in figure 3 g). Finally, we are left with a single
native contamination.
Further deviation possibilities are explored for net a to avoid
single contamination remained so far as shown in figure 3 h). The
result even worsens with the single native contamination remaining
as it is with an increase in distance of a. Hence the results accepted
for the path estimation is displayed clearly in figure 3 i).
The route implementation with latest arrival times and the requisite
stalls is shown in figure 3 j). Table 1 represents the result for the
route performance.
The latest arrival time(LAT) is computed = 25 units and electrode /
cell utilization = 83. Remaining native contaminations =1.

Table 1. Route performance table

Fig.4. Bar graph comparison for the number of

contaminations after route implementation

VI . EXPERIMENTAL RESULTS
Benchmark suite III comprised of four testbenches are used for
simulation of route performance deploying the proposed application
of Route Zone based technique. The esteemed testbenches are
namely -In-vitro1, In-vitro 2, Protein-1 and Protein-2 – each one is
constituted of a number of subproblems constituting of one bioassay
for one testbench each. Table 2 computes the results of simulations
using 4 testbenches presenting standard parameters for route Fig 5. Bar graph comparison for the total arrival time after
performances - maximum arrival time, average arrival time, number route implementation
of contaminations and the cells or electrodes utilization. Table 3
compares Bioassay execution performance in terms of droplet
transportation applying the method proposed and the contemporary
state of the art contributions.
It is observed that for both – longest arrival time and cell
utilization parameters - proposed method generates better outcome.
Figures 4, 5 and 6 display the bar graphs for comparison of No of

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Fig. 6. Bar graph comparison for the Cell/Electrode Utilization after
route implementation
VII. CONCLUSION
In this work, we have introduced a novel approach for efficient and
error free routing of heterogeneous droplets. The primary objective
of this work is to minimize the occurrence of intersections among
the droplets, which effectively results into minimization of required
wash droplets for residual or native contaminations. This innovative
technique establishes a designated routing zone for each network
and determines the preferred course of action when encountering
intersections. The ultimate goal is to optimize the overall
performance of the routing process by considering factors such as
contaminations, the timing of droplet arrivals, and efficient
utilization of cells. Through simulations using benchmark suite III,
we have observed enhanced routing performance with our proposed
method. However, it is worth noting that the effectiveness of this
technique may be diminished when applied to layouts involving a
significantly larger number of networks which needs further
exploration of intelligent pathfinding techniques.
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