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How to conduct a Thrombosis Assay using Cellix's VenaFlux Basic setup with ExiGo pump

Introduction

VenaFlux Solutions mimic human blood vessels

The circulatory system contains many different types of blood vessels which delivers blood cells, nutrients, oxygen, carbon dioxide and hormones to all parts of the body. Blood vessels play a huge role in many medical conditions such as:

  • Thrombosis

  • Inflammation

  • Atherosclerosis

VenaFlux Solutions comprise packages to suit all budgets, with our Starter kit including a precision microfluidic pump, a microenvironmental chamber to hold the biochips which contain engineered micro-capillaries and cell analysis software; to Pro and Elite versions including high-end fluorescent microscopes with a digital camera. The VenaFluxAssay software integrates the microfluidic pump, microscope and camera resulting in a complete automated high-throughput flow assay system for shear stress experiments.


Conducting a Thrombosis Assay using Cellix's VenaFlux Basic setup with ExiGo pump in four simple steps:


STEP 1

Initialise SmartFlo and select syringe

  • Initialising SmartFlo Software enables control of the ExiGo Pump.

  • Open the SmartFlo Software program and double click on the ExiGo Pump to initialise the pump.

  • Double click the syringe to open the "Syringe Database."

  • Select the one mil syringe and click "Set."


STEP 2

ExiGo Pump set-up which involves which involves placing the syringe with the blood sample into the pump

  • Using a 1mL syringe aspirate the blood sample from the eppendorf tube.

  • Ensure that the bubbles are removed by withdrawing sample, gently tapping and dispensing sample until it reaches the tip of the syringe

  • Continue aspirating the blood sample until the syringe is full.

  • Connect lure with tubing to the tip of the syringe.

  • Place output of tubing in a collection vessel and gently dispense sample to refill tubing with blood.

  • Place the syringe in the ExiGo Pump and use the SmartFlo software to drag and drop so that the plunger sits in place.

  • Turn the lead screw to secure the plunger in place.

  • Place the ExiGo syringe clamp and screw to secure the syringe.


STEP 3

Biochip Preparation

  • Place the pre-coated biochip in the microenvironmental chamber.This heats the chip to 37 °C suitable for cell based assays.

  • To prime the channels of the biochips, first pre-wet the input and the output ports of the biochip to prevent evaporation.

  • This is achieved by pipetting 40μL of PBS into the input and output ports of the chip.

  • Ensure the liquid is pipetted in to the microwells and not the channel of the chip.

  • An optional step is to pre-wash each channel of the biochip with PBS.


STEP 4

Run a whole blood thrombosis assay

  • Place the eppendorf and connect to chip via tubing.

  • Start the flow rate of the ExiGo pump at 10μL /min to ensure the tubing is completely pre-filled and the blood sample has started flowing from the out of the tubing before connecting to the chip.

  • After starting blood flow, connect tubing from ExiGo to input of chip.

  • Now review the sheer stress table and select the flow rate corresponding to the sheer stress for the biochip you are using.

  • In this example, we are using a Vena8 Fluoro+ Biochip and running at a sheer stress of 67.5 dyne/cm2 or a sheer rate of 1500 per second so we select a Flow Rate of 60μL /min.

  • Input the flow rate into the SmartFlo Software of the ExiGo Pump and Click Run.

  • Open the digital camera and attach to your microscope so you can take images and videos.

  • Thrombi formation can be seen in the biochip as shown below.


How to get started?

Thinking about trying out similar experiments in your lab? Here's what you'll need as a minimum setup:

  • VenaFlux Basic platform – a semi-automated microfluidic platform designed for conducting studies on cell adhesion, binding, rolling, and migration under shear flow conditions that replicate in vivo flow rates.

  • Vena8 Fluoro+ Biochips – to mimic the shear stress of arterial circulation.

  • ExiGo Microfluidic Pump – a pulse-free syringe pump for low-flow microfluidic applications.

  • Microenvironmental chamber – a temperature-controlled frame that keeps the biochip at 370˚ C.

  • Inverted microscope – we supply the Zeiss AxioVert A1 with the VenaFlux Pro option or the Zeiss AxioObserver7 with the VenaFlux Elite option.

  • Digital camera – to capture images and video recordings. We supply the Prime BSI Express with the VenaFlux Pro and Elite options.

  • Image Pro Cell Analysis software – for image and video analysis.


If you already have some of these items (such as the inverted microscope, camera, or cell analysis software), we recommend the VenaFlux Starter Basic kit. We have options that suit all budgets. You can check them out on our eShop.


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