Research

Facilities

Cell Culture Room

The cell culture laboratory has dedicated vertical laminar flow hoods for primary and immortalised cell culture. Stem cell isolation is carried out in self-contained tissue culture room.

 

Microscopy Room

The Microscopy Room at RDU@NHC amongst others, also houses a Carl Zeiss LSM 510 NLO laser confocal microscope. This confocal system is equipped with 3 laser lines (488nm, 543nm and 643nm) as well as an NLO laser, allowing for high resolution imaging, Ca²⁺ spark detection as well as deep tissuing imaging. Other microscopes also include a Leica stereomicroscope and two Carl Zeiss LSM 200 widefield inverted microscopes, one of which is fully motorised and uses either the CoolSNAP HQ or the high speed Cascade 512B camera to capture images.

 

Histology

RDU’s histology section is furnished with comprehensive facilities for cryo, paraffin and plastic resin sample preparation. Cryo samples are embedded in tissue freezing medium prior to sectioning using the Leica CM1850 refrigerated microtome. In contrast, paraffin samples are processed, embedded and sectioned using the Leica RM2135 microtome. Immunohistological tissue analyses can be performed after the desired tissue thickness is achieved by the sectioning.

In addition, RDU has expertise in stent histomorphometry using Leica SP1600 saw microtome and Leica SP2600/SM2500 ultramilling microtome. This setup is also suitable for hard and brittle materials using plastic resin embedding method.

 

 

Flow Cytometry

NHCS RDU houses a state-of-the-art BD FACSAria II Cell Sorter equipped with 3 built-in lasers(488nm blue laser, 633nm red laser and 355nm UV laser). Its revolutionary optics system consist of octagon and trigon-shaped collection devices which allow simultaneous detection of 15 parameters. This high-speed sterile cell sorter has the ability to perform plate sorting, slide sorting and up to 4-way tube sorting to fully meet the challenges of demanding scientific questions.

With our in-house expertise in flow cytometry, NHCS RDU also offers flow acquisition and cell sorting services to external users.

 

Cardiac Electrophysiology

RDU has acquired the Micro Electrode Arrays (MEA) system from Multichannel Systems that served as a good platform for cardiac electrophysiological studies. The MEA allows simultaneous cultivation and recording of the extracellular field potentials (FPs) of the spontaneous contracting cardiac samples at 37oC with the temperature control. This enables the electrophysiological and pharmacological characterization of cardiomyocytes derived from human induced pluripotent stem cells. Hence, the cardiomyocytes cultured on the MEA is a valuable assay in regenerative medicine and also in cardiac toxicity and safety pharmacology testing such as QT prolongation and arrhythmia. FPs recording with the MEA system can be combined with other techniques such as intracellular calcium imaging and patch clamp. 

 

Patch-Clamp Electrophysiology

Whole-cell patch-clamp recordings were conducted using an amplifier (Axon Instrument, Axopatch 200B) and an inverted microscope equipped with differential interface optics (Nikon, TE2000-S). The glass pipettes were prepared using borosilicate glass (Sutter Instrument, B150-86-10) using a micropipette puller (Sutter Instrument, Model P-97). The patch-clamped cell was superfused by means of a temperature controlled micro-superfusor (Warner instrument, TC-324B). Voltage-clamp measurements were conducted to record multiple ion channel of cardiomyocytes generated from iPSCs. Current-clamp recording were conducted to recording action potential and classify the different cardiac subtypes. A pClamp software (Axon Instrument , Clampex 10.0) was used to generate voltage-pulse protocols, acquire and analyze data.

 

Anton Paar Rheometer MCR101

Anton Paar Rheometer is a versatile machine in recording and analyzing the mechanical stimulations and responses of various soft materials. It gives results such as complex tensile modulus, complex shear modulus, ultimate strength, etc. which are relevant for our application in cardiac tissue engineering. Based on the results, we study the cellular responses of the cells seeded onto scaffolds of varying modulii; and then design a scaffold with compatible modulus for engineered tissue patch.

 

 

Cardiac Ultrasound

RDU has recently equipped with VisualSonics Vevo 2100 imaging system and high frequency transducer central frequency 30mHz, axial resolution up to 50µm), which is specially designed for small animal with ever better image quality as compared to usual ultrasound machine (axial resolution usually at 200µm for small animal). It provides real time assessment of haemodynamic changes and cardiovascular function. Therefore, it allows researchers to assess the progress of cardiovascular disease models such as, myocardial infarction, diastolic dysfunction, cardiac hypertrophy and valvular heart disease. Beside the conventional modalities such as 2D, m-mode, color doppler and pulse wave doppler, Vevo 2100 provides more complication analysis such as tissue doppler imaging and strain study. Vevo 2100 also allows physiological parameters such as body temperature, ECG, heart rate and respiration to be monitored throughout an imaging session. In additional, it allows user to capture images by hand free method with the use of its Integrated Rail System.

Research and Development Unit is also equipped with GE Vivid 7 ultrasound system for big animal imaging.