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Molecular, Imaging & Analytical Core Facility

The Molecular, Imaging, and Analytical Core facility contributes greatly to the research projects conducted by the THRCE & COBRE investigators and provides a comprehensive service for molecular biology, imaging and biochemistry support. This facility serves as the resource to instruments and equipment needed to perform advanced molecular biology, semi-quantitative immuno-histochemistry and bio-analytical experiments, and support training of junior faculty, postdoctoral fellows, graduate and medical students in the areas of physiology, hypertension and cardiovascular and renal diseases.

FACILITIES & RESOURCES
The Molecular and Imaging Core Laboratory consists of the following laboratories:

1. Main laboratory

1-1. DNA workspace
This space is dedicated for DNA work such as genotyping of gene-targeted mice and analyzing for promoter regions of target genes. A well-trained laboratory research technician handle samples. He/She will extract genomic DNA from the snipped tails of gene-targeted mice and perform PCR. He/She will also set up agarose gel electrophoresis and report the final results to the corresponding investigators.

  • Applied Biosystems GeneAmp PCR System,
  • Fisher Scientific’s dual chamber water bath,
  • Sanyo’s microwave,
  • Fisher Scientific’s power supply,
  • Bio Rad’s horizontal agarose gel-electrophoresis system,
  • UVP’s ultraviolet transilluminator,
  • Fisher Scientific’s portable centrifuge machine, and
  • New Brunswick Scientific’s incubator shaker.
  • Biotrack II Plate washer & Jitterbug plate incubator/shaker

1-2. Western blot area
This space is dedicated for Western blot analysis. A well-trained laboratory research technician will handle samples. He/She will extract protein samples from human/rats/mouse samples or cells provided by the investigators. He/She will also set up polyacrylamide gel electrophoresis and report the final results to the corresponding investigators.

  • Licor’s Odyssey system,
  • Pharmacia Biotech’s power supply and
  • Novex’s vertical PAGE system.
  • Alpha Innotech Fluorechem

1-3. Shared open space
This open space is shared as a pipetting station.

  • Eppendorf’s bench-top refrigerated centrifuge machine and
  • Denver Instrument’s high quality scale.

1-4. Histology/Immunohistochemistry workspace
This space is dedicated for histology/immunohistochemistry works. A well-trained laboratory research technician will handle samples. He/She will set up slide glasses of human/rats/mouse samples provided by the investigators. He/She will also perform histology and/or immunohistochemistry staining and return the stained slides to the corresponding investigators. 

  • Dako Cytomation’s Autostainer for immunohistochemistry robot,
  • Fisher Scientific’s chemical hume hood for straining,
  • Fisher Scientific’s slide oven,
  • Thermolyne’s slide shaker, and
  • Fisher Scientific’s dry incubator.

1-5. ELISA station
This space is dedicated for ELISA work. A well-trained laboratory research technician will handle samples. He/She will prepare protein samples for ELISA from human/rats/mouse samples provided by the investigators. He/She will also set up ELISA plates and report the final results to the corresponding investigators.

  • Amersham Biosciences’ ELISA plate washer,
  • Boekel Scientific’s ELISA plate shaker/incubator,
  • BMG Labtech's FLUOstar Optima multi-detection microplate reader, and
  • Bayer’s DCA2000 creatinine/albumin auto-analyzer.

1-6. RNA workspace
This space is dedicated for RNA work such like RNA extraction and real-time RT-PCR. A well-trained laboratory research technician will handle samples. He/She will extract total RNA samples from human/rats/mouse samples or cells provided by the investigators using Qiagen’s robotic system. He/She will also set up real-time RT-PCR and report the final results to the corresponding investigators.

  • Qiagen’s BioRobot EZ1,
  • Amersham Biosciences’ GeneQuant spectrometer,
  • Fisher Scientific’s portable centrifuge machine and
  • Fisher Scientific’s tissue homogenizer.

1-7. Analytical equipment station
This space is dedicated for analytical equipment.

  • Stratagene’s real-time PCR machine Alphainotech’s densitometric machine, and
  • Licor’s Odyssey system.

Major Equipment

A) Licor’s Odyssey system
The Odyssey Infrared Imaging System (LI-COR) uses two completely independent detection channels - one for each infrared dye. Two diode lasers provide excitation light at 685 and 785 nm. Two avalanche photodiodes are filtered to detect fluorescence at 720 and 820 nm. Scans can be performed at resolutions ranging from 21 - 337 µm. Two detection channels allow us to probe two separate targets in the same experiment. Quantification accuracy is increased when the second color is used for normalization. Odyssey provides a flexible, multifunctional platform to accommodate a variety of applications such as ultra-rapid Western blots, in-gel Western, in-cell Western, ELISA/FLISA, electromobility shift assays, Northern blots, Southern blots, and in vivo imaging.
 

B) BMG Labtech's FLUOstar Optima multi-detection microplate reader
BMG Labtech's FLUOstar Optima multi-detection microplate reader can take care of:

  • UV/Visible absorbance: ELISA, RNA determination, etc.
  • Luminescence: Dual Luciferase assay
  • Fluorescence: GFP, intracellular calcium, etc.

 

C) Qiagen’s BioRobot EZ1
The BioRobot EZ1 workstation (Qiagen) purifies high-quality nucleic acids (DNA or RNA) from tissues as well as from cells. From 1 to 6 samples are processed in 15-20 minutes. The rapid speed of nucleic acid purification is achieved through magnetic particle technology: nucleic acids in sample lysates are isolated in one step through their binding to the silica surface of magnetic particles. Reagents are supplied in pre-filled EZ1 Reagent Cartridges, which ensures speed and convenience in loading the BioRobot EZ1. Protocols for nucleic acid purification are supplied on pre-programmed EZ1 Cards.
 

These protocols provide both on-screen instructions for the operator and operating commands for the workstation. The range of kits and protocols for the BioRobot EZ1 is continuously expanding to allow nucleic acid purification from a wide range of sample types.
 

D) Dako Cytomation’s Autostainer
Dako Cytomation’s Autostainer for immunohistochemistry robot can take care of up to 48 slices in one run, which is required to do immunohistochemistry in a semi- quantitative manner. Moreover, we can apply individual conditions on each slide - different antibody concentration and different incubation time, which saves significant time to optimize the immunohistochemistry condition.
 


2. Cell culture workroom
A separated room is dedicated for cell cultures.

  • Labconco’s purifier class II biosafety cabinet Heraeus’s cell incubator, and
  • Fisher Scientific’s inverted microscope.

Major Equipment

A) Nexcelom Cellometer Automated Cell Counter
Nexcelom's Cellometer is an automated cell counter, which allows us to count cells, determine viability, measure cell sizes, etc. all with a click of a button in less than 30 sec. This set up will improve data quality, and provide additional data such like cell sizes and cell images, which can ultimately improve the research quality.
 

B) Next Advance Inc.'s Bullet Blender BLUE
Next Advance Inc.'s Bullet Blender BLUE is the homogenizer. It makes the homogenized samples from cells and tissues for protein, RNA or DNA work in a few minutes with air cooling.
 


3. Microscope Workroom
A separate room dedicated for microscope and image analyses. 

  • Olympus’s (general) microscope equipped to real-time digital image capturing system,
  • Olympus’s fluorescence-ready microscope equipped to real-time digital image capturing system, and
  • Image-Pro plus image handling/analyzing system.

Major Equipment

EVOS fl Fluorescence Microscope
EVOS fl digital inverted microscope combines advanced ergonomic design, an on-board microprocessor, LED illumination, fully integrated high-sensitivity fluorescence camera, and a highly streamlined user interface to deliver unprecedented flexibility and ease-of-use. EVOS fl on-board computer lets us capture and manage multi- channel fluorescence image data right on the microscope through "insanely" easy to use software.
 


4. RIA and Chemistry Core Labs

These two labs allow for the quantification of various parameters from various tissues, like plasma, tissues, and urine.  

  • HPLC separation of various angiotensin peptides from plasma and tissue extracts
  • Detection of radioactivity signal from specific antibody-isotope-labelled ligand complexes
  • Assessment of millimolar concentrations of Na+ and K+ in plasma and urine
  • Measurement of millimole/kg concentrations of solutes in plasma and urine samples
  • Quantification of fluorescence signals from ATP assays and Luciferase assays
  • Calculation of concentrations of unknowns for a given enzyme-linked immunoanalysis assay

Some of the major pieces of equipment include:

  • Waters 600 controller, 2487 Dual X absorbance detector, 2475 Multi X fluorescence detector, and ISCO Foxy fraction collector
  • Packard Multiprobe II EX robotic liquid handling system
  • Wallac Wizard 1470 Automatic Gamma Counter
  • Instrumentation Labs IL 943 Flame Photometer with an automatic sampler
  • Wescor Vapro Vapor Pressure Osmometer
  • Turner Biosystems 20/20n Luminometer
  • Amersham Biosciences Biotrak II Plate Reader.

Major Equipment

A) Thermo Scientific's NanoDrop 2000
Thermo Scientific's NanoDrop 2000 is a UV/Vis spectrometer that allows for the quantification of DNA, RNA, protein and peptide samples. Because it needs only 0.5 ul of sample for measurement, we can save the volume of samples. Concentration measurement capability is from 2 to 15,000 ng/ul (dsDNA), which eliminates the need to dilute samples. The measurement time is less than 5 seconds to each sample.
 

B) Fisher Scientific's accuSpin Micro17R Microcentrifuge
Fisher Scientific's accuSpin Micro17R Microcentrifuge is a refrigerated microcentrifuge. It can centrifuge 1.5 ml or 2.0 ml tubes up to 24 at a time. The cooling system preserves the sample quality.