Dynamics of a two-layer immiscible fluid system exposed to ultrasound.

The relocation dynamics of a two-layer immiscible fluid system exposed to bulk acoustic waves using simulations and experiments are reported. A theoretical formulation of the acoustic radiation pressure (ARP) acting on the interface reveals that ARP is a nonlinear function of the impedance contrast. It has been shown that the force acting on the interface is the simple sum of the ARP and the interfacial tension, which is dependent on the angle of the interface. It was discovered that although the acoustic radiation force is directed from high-impedance fluid (HIF) to low-impedance fluid (LIF), the final steady-state configuration depends on the wall-fluid contact angle (CA). Our study reveals that the HIF and LIF would relocate to the channel center for CA>110°, and CA<70°, respectively, while complete flipping of the fluids is observed for intermediate angles. The forces relocate the fluids in the channel, generally, by a clockwise or anticlockwise rotation. Here, it is demonstrated that the direction of this twist can be determined by the relative densities and wettabilities of the two fluids.

Ultrasound resonance in coflowing immiscible liquids in a microchannel.

We study ultrasonic resonance in a coflow system comprising a pair of immiscible liquids in a microchannel exposed to bulk acoustic waves. We show using an analytical model that there are two resonating frequencies corresponding to each of the coflowing liquids, which depend on the speed of sound and stream width of the liquid. We perform a frequency domain analysis using numerical simulations to reveal that resonance can be achieved by actuating both liquids at a single resonating frequency that depends on the speeds of sound, densities, and widths of the liquids. In a coflow system with equal speeds of sound and densities of the pair of fluids, the resonating frequency is found to be independent of the relative width of the two streams. In coflow systems with unequal speeds of sound or densities, even with matching characteristic acoustic impedances, the resonating frequency depends on the stream width ratio, and the value increases with an increase in the stream width of the liquid with a higher speed of sound. We show that a pressure nodal plane can be realized at the channel center by operating at a half-wave resonating frequency when the speeds of sound and densities are equal. However, the pressure nodal plane is found to shift away from the center of the microchannel when the speeds of sound and densities of the two liquids are unequal. The results of the model and simulations are verified experimentally via acoustic focusing of microparticles suggesting the formation of a pressure nodal plane and hence a resonance condition. Our study will find relevance in acoustomicrofluidics involving immiscible coflow systems.

Effects of surface acoustic waves on droplet impact dynamics.

HYPOTHESIS: Surface acoustic waves (SAW) propagating along a solid surface can significantly affect the dynamics of droplet impact. Although droplet impact in presence of SAW has been attempted recently, here, we investigate the effects of surface wettability, droplet size, impact velocity, and SAW power on the impact and spreading dynamics along with post-impact oscillation dynamics of a drop. EXPERIMENTS: Here, we study droplet impact on a surface exposed to traveling SAW produced using an interdigitated electrode patterned on a piezoelectric substrate. The effects of Weber number (We), surface wettability, and SAW power on the impact and spreading dynamics and post-impact oscillation dynamics are studied. FINDINGS: Our study unravels that the interplay between capillary and viscous forces, and inertia forces arising due to pre-impact kinetic energy and SAW-induced bulk acoustic streaming underpins the phenomena. Remarkably, we find that the effect of SAW on droplet impact dynamics is predominant in the case of a hydrophilic (HPL) substrate at a higher SAW power and smaller We and hydrophobic (HPB) substrate irrespective of SAW power. Our study reveals that the maximum droplet spreading diameter increases with SAW power at smaller We for an HPL surface whereas it is independent of SAW power at higher We. Post-impact oscillation of a droplet over an HPL surface is found to be overdamped with a smaller amplitude compared to an HPB substrate, and a faster decay in oscillation amplitude is observed in the case of an HPB surface and higher We. Our study provides an improved understanding of droplet impact on a surface exposed to SAW that may find relevance in various practical applications.

Deicing of Sessile Droplets Using Surface Acoustic Waves.

Deicing has significant relevance in various applications such as transportation, energy production, and telecommunication. The use of surface acoustic waves (SAWs) is an attractive option for deicing as it offers several advantages such as localized heating, in situ control, low power, and system integration for highly efficient deicing. Here, we report an understanding of the dynamics of deicing of microlitre volume water droplets (1 to 30 µL) exposed to low power (0.3 W) SAW actuation using an interdigitated electrode on a piezoelectric (LiNbO3) substrate. We study the time variation of the volume of liquid water from the onset of SAW actuation to complete deicing, which takes 2.5 to 35 s depending on the droplet volume. The deicing phenomenon is attributed to acoustothermal heating which is found to be greatly influenced by the loss of ice adhesion with the substrate and the acoustic streaming within the liquid water. Acoustothermal heating inside the droplet is characterized by the temperature distribution inside the droplet using infrared thermography, and acoustic streaming is observed using dye-based optical microscopy. A rapid enhancement in deicing is observed upon the detachment of ice from the substrate and the onset of acoustic streaming, marked by a sudden increase in the liquid water volume, droplet temperature, and heat transfer coefficient. The deicing time is found to increase linearly with droplet volume as observed from experiments and further verified using a theoretical model. Our study provides an improved understanding of the recently introduced SAW-based deicing technique that may open up the avenue for a suitable alternative to standard deicing protocols.

Microfluidics-based rapid measurement of nitrite in human blood plasma.

Nitric oxide (NO) is one of the vital gasotransmitters that takes part in many biological pathways such as infection, inflammation and ischemia, immune response, neurotransmission, and cardiovascular systems. Nitrite is one of the primary metabolites of NO and is considered to be a circulating storage pool for NO. Here, we report direct and rapid measurement of nitrite in human blood plasma using a fluorescence-based microfluidic method. The study revealed the factors that affect the endogenous concentration of nitrite in blood plasma, mainly the presence of blood cells, hemoglobin, and soluble proteins. We find that separation of blood plasma immediately after sample collection and subsequent dilution of plasma with buffer at a ratio of 1 : 4 eliminates the interference from cells and proteins, providing reliable measurements. The proposed method can measure plasma nitrite in the concentration range of 0-20 µM with a limit of detection of 60 nM and a sensitivity of 5.64 µM-1 within 10 min of sample collection. By spiking nitrite into plasma, a linear correlation between the nitrite concentration and FL intensity is obtained, which is utilized further to measure the endogenous concentration of nitrite present in the plasma of healthy volunteers and patients. The study revealed that the endogenous nitrite concentration in the blood plasma of healthy humans falls in the range of 0.4-1.2 µM. Furthermore, the study with blood samples obtained from patients showed that nitrite levels are inversely correlated with the total cholesterol and low-density lipoproteins levels, which is in good agreement with the literature.

Acoustic particle trapping driven by axial primary radiation force in shaped traps.

We study particle trapping driven by the axial primary radiation force (A-PRF) in shaped traps exposed to standing bulk acoustic waves (S-BAW) using numerical simulations and experiments. The utilization of the stronger A-PRF as the main retention force is a consequence of standing-wave formation along the flow direction, instead of the orthogonal direction as in the case of traditionally used lateral-PRF S-BAW trapping setups. The study of particle dynamics reveals that the competition between A-PRF and viscous drag force governs particle trajectory. The ratio of the acoustic energy to the viscous work (ß) provides a general criterion for particle trapping at a distinctive off-node site that is spatially controllable. Particles get trapped for ß≥ß_{cr} at some distance away from the nodal plane and the distance varies as ß^{-c} (c=0.6-1.0). The use of A-PRF as the retention force could potentially allow traditional S-BAW trapping systems to envisage high-throughput advancements surpassing the current standards in cell-handling unit operations.

Trapping of Aqueous Droplets under Surface Acoustic Wave-Driven Streaming in Oil-Filled Microwells.

Microwell arrays are ideal platforms for cell culturing, cell separation, and low-volume liquid handling. The ability to manipulate droplets in microwells could open up the opportunity for developing new biochemical assays. Here, we study the trapping of aqueous droplets in an oil-filled microwell driven by the application of nanometer amplitude vibrations called surface acoustic waves (SAW). We elucidate the dynamics of the droplet within the vortex toward the final trapping location and the physics of the trapping phenomenon using a theoretical model by considering the relevant forces. Our study revealed that the combined effect of acoustic radiation and hydrodynamic forces leads to droplet migration and trapping. We demarcate the trapping and nontrapping regimes in terms of the minimum critical input power required for the trapping of droplets of different sizes and densities. We find that the critical power varies as the square of the droplet size and is higher for a denser droplet. The effects of input power and droplet size on the trapping location and trapping time are also studied.

Chemical analysis of low carbon content coals and their applications as dye adsorbent.

Coal is primarily a fuel material but lately it has been utilized as an adsorbent for removing toxic metal ions. However, its usage for removing organic pollutants is not well studied. We report here a systematic study on the use of coal samples of varying carbon contents as adsorbents for removing Basic Blue 41 as a model cationic dye. The coal samples were collected from coal mines and were thoroughly characterized. The concentrations of carbon, hydrogen, oxygen, nitrogen and sulphur contents were measured by CHNS analyzer. The concentrations of aluminum, silicon, sulphur, titanium and iron were determined by EDXRF, which corresponded to silicon dioxide (quartz) and aluminium silicate (kaolinite) as the major mineral inclusions, corroborated by XRD results and micrographs showing elemental maps determined from SEM-EDAX. The coal samples with low carbon content revealed higher adsorption capacity (qe âˆ¼ 8.0-9.3 mg/g) of Basic Blue dye at optimized adsorbent dose (2 mg/mL), pH 9 and contact time (120 min). The adsorption kinetic studies satisfied pseudo second order model and the intra-particle diffusion of the dye was evident. The dye adsorption followed Langmuir adsorption isotherm, and the qmax values ranged between 17 and 30 mg/g for low carbon content coal. The FT-IR, Brunauer-Emmett-Teller (BET) surface area and zeta potential results of the coal samples could explain the adsorption phenomenon of cationic dye. The kinetic and thermodynamic studies revealed that the adsorption of Basic Blue 41 dye was based on chemisorptions mechanism.

Elastocapillary interaction between a long rectangular membrane and a liquid drop.

We report elastocapillary interaction between a long rectangular membrane fixed along its central axis and a liquid drop dispensed at one of its ends. The introduction of the drop results in the elastocapillary-driven wrapping of the membrane along its width and a concomitant flow in the resulting conduit along its length. Depending upon the drop size (d) and capillary length scale (Lc), we identified general criteria for achieving complete wrapping of the membrane in the dry state from energy considerations. For small droplets satisfying d ≲ Lc, we find that the critical membrane length (Wc) required for complete wrapping is proportional to the elastocapillary length scale (Lec). In the case of large droplets with d > Lc, the wrapping behavior depends on the ratio of membrane width to elastocapillary length scale (W/Lec) and the ratio of capillary length scale to the elastocapillary length scale (Lc/Lec). Our study suggests that the critical membrane width for complete wrapping is smaller in the wet state compared to that in the dry state, which can be attributed to the existence of a transmembrane pressure in the wet state. The effect of membrane thickness and width and drop volume on the length and cross-section of the wrapped conduit and attached width of the wrapped membrane is studied. For small droplets, the resulting elastocapillary flow exhibits an inertial regime at small times, followed by a Washburn regime at intermediate times, and finally an inertial regime, and for large droplets, only an inertial regime is observed throughout.

Reversible Stream Drop Transition in a Microfluidic Coflow System via On Demand Exposure to Acoustic Standing Waves.

Transition between stream and droplet regimes in a coflow is typically achieved by adjusting the capillary numbers (Ca) of the phases. Remarkably, we experimentally evidence a reversible transition between the two regimes by controlling exposure of the system to acoustic standing waves, with Ca fixed. By satisfying the ratio of acoustic radiation force to the interfacial tension force, Ca_{ac}>1, experiments reveal a reversible stream drop transition for Ca<1, and stream relocation for Ca≥1. We explain the phenomenon in terms of the pinching, advection, and relocation timescales and a transition between convective and absolute instability from a linear stability analysis [P. Guillot et al., Phys. Rev. Lett. 99, 104502 (2007)PRLTAO0031-900710.1103/PhysRevLett.99.104502].

Dynamical motion of a pair of microparticles at the acoustic pressure nodal plane under the combined effect of axial primary radiation and interparticle forces.

The dynamical motion of a pair of microparticles exposed to acoustic standing waves and located at the pressure nodal plane is studied using numerical simulations and experiments. The insight into their dynamical behavior along the pressure nodal plane due to the competition between the axial primary radiation and interparticle forces is elucidated. An expression for axial primary radiation force acting on a particle is derived, and the particle dynamics is simulated using fluid-structure interaction model based on the arbitrary Lagrangian-Eulerian method. Considering the total radiation force acting on a particle is the sum of the axial primary radiation force and the interparticle force, three distinct dynamical regimes are observed depending upon the relative magnitudes of the acoustic forces which in turn depend on the gradient of the acoustic energy density. Acceleration, deceleration, and constant velocity motion of the pair of approaching particles are observed, which are explained by the interplay of the acoustic and non-acoustic forces. The dynamical motion of the pair of particles predicted using the model is in very good agreement with the experimental observations.

Particle encapsulation in aqueous ferrofluid drops and sorting of particle-encapsulating drops from empty drops using a magnetic field.

Handling and manipulation of particle-encapsulating droplets (PED) have profound applications in biochemical assays. Herein we report encapsulation of microparticles in aqueous ferrofluid droplets in a primary continuous phase (CP) and sorting of PED from empty droplets (ED) at the interface of the CP in coflow with a second continuous phase using a magnetic field. We find that the encapsulation process results in a size contrast between the PED and ED that depends on the flow regime - squeezing, dripping, or jetting - which in turn is governed by the ratio of the discrete phase to the continuous phase capillary number, Car. The difference between the volume fractions of ferrofluid in the PED and ED, ΔαPED, is utilized for sorting, and is found to depend on the ratio of the capillary numbers, Car. The difference ΔαPED is found to be maximum in the jetting regime, suggesting that the jetting regime is most suitable for encapsulation and sorting. The sorting criterion is represented in terms of a parameter ξ, which is a function of the ratios of the magnetic force to the interfacial force experienced by the PED and ED. Our study revealed that sorting is possible for ξ < 0, which corresponds to ΔαPED > 0.25. The maximum sorting efficiency of our system is found to be ∼95% at a throughput of ∼100 drops per s.

Direct and rapid measurement of hydrogen peroxide in human blood using a microfluidic device.

The levels of hydrogen peroxide ([Formula: see text]) in human blood is of great relevance as it has emerged as an important signalling molecule in a variety of disease states. Fast and reliable measurement of [Formula: see text] levels in the blood, however, continues to remain a challenge. Herein we report an automated method employing a microfluidic device for direct and rapid measurement of [Formula: see text] in human blood based on laser-induced fluorescence measurement. Our study delineates the critical factors that affect measurement accuracy-we found blood cells and soluble proteins significantly alter the native [Formula: see text] levels in the time interval between sample withdrawal and detection. We show that separation of blood cells and subsequent dilution of the plasma with a buffer at a ratio of 1:6 inhibits the above effect, leading to reliable measurements. We demonstrate rapid measurement of [Formula: see text] in plasma in the concentration range of 0-49 µM, offering a limit of detection of 0.05 µM, a sensitivity of 0.60 µM-1, and detection time of 15 min; the device is amenable to the real-time measurement of [Formula: see text] in the patient's blood. Using the linear correlation obtained with known quantities of [Formula: see text], the endogenous [Formula: see text] concentration in the blood of healthy individuals is found to be in the range of 0.8-6 µM. The availability of this device at the point of care will have relevance in understanding the role of [Formula: see text] in health and disease.

Coalescence of Droplets in a Microwell Driven by Surface Acoustic Waves.

Microwell arrays are amongst the most commonly used platforms for biochemical assays. However, the coalescence of droplets that constitute the dispersed phase of suspensions housed within microwells has not received much attention to date. Herein, we study the coalescence of droplets in a two-phase system in a microwell driven by surface acoustic waves (SAWs). The microwell structure, together with symmetric exposure to SAW irradiation, coupled from beneath the microwell via a piezoelectric substrate, gives rise to the formation of a pair of counter-rotating vortices that enable droplet transport, trapping, and coalescence. We elucidate the physics of the coalescence phenomenon using a scaling analysis of the relevant forces, namely, the acoustic streaming-induced drag force, the capillary and viscous forces associated with the drainage of the thin continuous phase film between the droplets and the van der Waals attraction force. We confirm that droplet-droplet interface contact is established through the formation of a liquid bridge, whose neck radius grows linearly in time in the preceding viscous regime and proportionally with the square root of time in the subsequent inertial regime. Further, we investigate the influence of the input SAW power and droplet size on the film drainage time and demarcate the coalescence and non-coalescence regimes to derive a criterion for the onset of coalescence. The distinct deformation patterns observed for a pair of contacting droplets in both the regimes are elucidated and the possibility for driving concurrent coalescence of multiple droplets is demonstrated. We expect the study will find relevance in the demulsification of immiscible phases and the mixing of samples/reagents within microwells for a variety of biochemical applications.

LSPR based on-chip detection of dengue NS1 antigen in whole blood.

The development of a biosensor for rapid and quantitative detection of the dengue virus continues to remain a challenge. We report a lab-on-chip device that combines membrane-based blood plasma separation and a localized surface plasmon resonance (LSPR) based biosensor for on-chip detection of dengue NS1 antigen from a few drops of blood. The LSPR effect is realized by irradiating UV-NIR light having a spectral peak at 655 nm onto nanostructures fabricated via thermal annealing of a thin metal film. We study the effect of the resulting metal nanostructures on the LSPR performance in terms of sensitivity and limit of detection, by annealing silver films at temperatures ranging from 100 to 500 °C. The effect of annealing temperature on the nanostructure size and uniformity and the resulting optical characteristics are investigated. Further, the binding between non-targeted blood plasma proteins and NS1-antibody-functionalized nanostructures on the LSPR performance is studied by considering different blocking mechanisms. Using a nanostructure annealed at 200 °C and 2X-phosphate buffer saline with 0.05% Tween-20 as the blocking buffer, from 10 µL of whole blood, the device can detect NS1 antigen at a concentration as low as 0.047 µg mL-1 within 30 min. Finally, we demonstrate the detection of NS1 in the blood samples of dengue-infected patients and validate our results with those obtained from the gold-standard ELISA test.

An optomicrofluidic device for the detection and isolation of drop-encapsulated target cells in single-cell format.

Single-cell analysis has emerged as a powerful method for genomics, transcriptomics, proteomics, and metabolomics characterisation at the individual cell level. Here, we demonstrate a technique for the detection and selective isolation of target cells encapsulated in microdroplets in single-cell format. A sample containing a mixed population of cells with fluorescently labelled target cells can be focused using a sheath fluid to direct cells in single file toward a droplet junction, wherein the cells are encapsulated inside droplets. The droplets containing the cells migrate toward the centre of the channel owing to non-inertial lift force. The cells present in the droplets are studied and characterised based on forward scatter (FSC), side scatter (SSC), and fluorescence (FL) signals. The FL signals from the target cells can be used to activate a selective isolation module based on electro-coalescence, using suitable electronics and a program to sort droplets containing the target cells in single-cell format from droplets containing background cells. We demonstrated the detection and isolation of target cells (cancer cells: HeLa and DU145) from mixed populations of cells, peripheral blood mononuclear cells (PBMC) + cervical cancer cells (HeLa) and PBMC + human prostate cancer cells (DU145), at a concentration range of 104-106 ml-1 at 300 cells per s. The performance of the device is characterised in terms of sorting efficiency (>97%), enrichment (>1800×), purity (>98%), and recovery (>95%). The sorted target cells were found to be viable (>95% viability) and showed good proliferation when cultured, showing the potential of the proposed sorting technique for downstream analysis.

Effect of surface energy and roughness on cell adhesion and growth - facile surface modification for enhanced cell culture.

In vitro, cellular processing on polymeric surfaces is fundamental to the development of biosensors, scaffolds for tissue engineering and transplantation. However, the effect of surface energy and roughness on the cell-surface interaction remains inconclusive, indicating a lack of complete understanding of the phenomenon. Here, we study the effect of surface energy (E s) and roughness ratio (r) of a polydimethylsiloxane (PDMS) substrate on cell attachment, growth, and proliferation. We considered two different cell lines, HeLa and MDA MB 231, and rough PDMS surfaces of different surface energy in the range E s = 21-100 mJ m-2, corresponding to WCA 161°-1°, and roughness ratio in the range r = 1.05-3, corresponding to roughness 5-150 nm. We find that the cell attachment process proceeds through three different stages marked by an increase in the number of attached cells with time (stage I), flattening of cells (stage II), and elongation of cells (III) on the surface. Our study reveals that moderate surface energy (E s ≈ 70 mJ m-2) and intermediate roughness ratio (r ≈ 2) constitute the most favourable conditions for efficient cell adhesion, growth, and proliferation. A theoretical model based on the minimization of the total free energy of the cell-substrate system is presented and is used to predict the spread length of cells that compares well with the corresponding experimental data within 10%. The performance and reusability of the rough PDMS surface of moderate energy and roughness prepared via facile surface modification are compared with standard T-25 cell culture plates for cell growth and proliferation, which shows that the proposed surface is an attractive choice for efficient cell culture.

Attraction and Repulsion between Liquid Droplets over a Liquid-Impregnated Surface.

We show that adjacent liquid droplets exhibit long-range attraction and repulsion on an immiscible liquid impregnating a surface when either the drop or the impregnating liquid is volatile. Remarkably, we find that at small times the interaction is attractive, analogous to the "Cheerios effect", but at large times the interaction becomes repulsive depicting the "reverse-Cheerios effect". Our study reveals that the interaction is underpinned by wetting and capillarity, buoyancy, and evaporation phenomena. We experimentally observe the interaction between a pair of droplets and provide a theoretical framework to quantitatively predict their transport behavior.

Continuous electrical lysis of cancer cells in a microfluidic device with passivated interdigitated electrodes.

Cell lysis is a critical step in genomics for the extraction of cellular components of downstream assays. Electrical lysis (EL) offers key advantages in terms of speed and non-interference. Here, we report a simple, chemical-free, and automated technique based on a microfluidic device with passivated interdigitated electrodes with DC fields for continuous EL of cancer cells. We show that the critical problems in EL, bubble formation and electrode erosion that occur at high electric fields, can be circumvented by passivating the electrodes with a thin layer (∼18 µm) of polydimethylsiloxane. We present a numerical model for the prediction of the transmembrane potential (TMP) at different coating thicknesses and voltages to verify the critical TMP criterion for EL. Our simulations showed that the passivation layer results in a uniform electric field in the electrode region and offers a TMP in the range of 5-7 V at an applied voltage of 800 V, which is well above the critical TMP (∼1 V) required for EL. Experiments revealed that lysis efficiency increases with an increase in the electric field (E) and residence time (tr): a minimum E ∼ 105 V/m and tr ∼ 1.0 s are required for efficient lysis. EL of cancer cells is demonstrated and characterized using immunochemical staining and compared with chemical lysis. The lysis efficiency is found to be ∼98% at E = 4 × 105 V/m and tr = 0.72 s. The efficient recovery of genomic DNA via EL is demonstrated using agarose gel electrophoresis, proving the suitability of our method for integration with downstream on-chip assays.

Understanding of the role of dilution on evaporative deposition patterns of blood droplets over hydrophilic and hydrophobic substrates.

Blood is a complex colloidal suspension which carries myriads of information about human health. Understanding the evaporation dynamics and its consequent deposition patterns have direct relevance in disease detection. We report evaporation dynamics of whole and diluted blood droplets over hydrophilic (glass) and hydrophobic (PDMS, polydimethylsiloxane) substrates. Our experiments show that blood drops evaporating on a hydrophilic substrate exhibit radial and orthoradial cracks in the coronal region and random cracks in the central region. Using Griffith's energy criterion, we show that crack formation takes place when the capillary pressure and the resulting compressive stress inside the evaporating droplet exceeds critical stress which depends on the elastic modulus, interfacial energy, and the particle concentration of the system. The width of the coronal region (w), the film thickness (h) at the contact line, and the crack pitch (p) decrease with increasing blood dilution. In the dilution range of 2.0-0.8% HCT (hematocrit), the transition from the cracking to the non-cracking regime is observed, which can be attributed to inadequate compressive stress available even after the evaporation of the blood droplet is completed. For the hydrophobic substrate, buckling instead of cracking is observed for the whole blood droplets, which can be attributed to the distinct wetting and evaporation kinetics. The buckling of the blood drop on a hydrophobic surface is attributed to the competition between capillary pressure originated due to the formation of an elastic network of RBCs (red blood cells) and the menisci formed between adjacent RBCs, and the critical buckling pressure. With increasing blood dilution, a transition from buckling (between 21 and 42% HCT) to cracking (between 21 and 2.0% HCT) of the droplets, and eventually to the non-cracking regime (between 2.0 and 0.8% HCT) is observed. Our study unravels the interesting attributes about one of the important physico-chemical factors (i.e. % HCT) that affect the evaporation of blood droplets and the resulting deposition patterns on substrates with different hydrophobicity.