Molecular Analysis

AFP “Mini Medical Series” X-Ray Film Processor

Description – Automated X-ray film processing for Westerns etc.

  • 4X4 inch (10X10cm) through 14X36 inch (35X91cm) film
  • Capable of processing up to 85 sheets per hour
  • Time and temperature adjustable for controlled development of film

Location: IRF Basement
IRF Contact: Jeff Davis

 

BioRad BioPlex

Description — The BioPlex system is a multiplex analysis system that permits the simultaneous analysis of up to 100 different biomolecules (proteins, peptides, or nucleic acids) in a single microplate well. By multiplexing with the Bio-Plex system, researchers can dramatically increase the amount of useful information from rare or volume-limited samples such as mouse or rat serum, and decipher complex interrelationships among proteins involved in signal transduction pathways. The microplate platform allows the automated analysis of 96-well plates with a throughput of more than 1,800 assay points (for a 23–plex assay) in 30 minutes. The Bio-Plex suspension array is built around three core technologies:

  • The first is the family of fluorescently dyed microspheres, or beads, from Luminex Corp.
  • The second is a flow cytometer with two lasers and associated optics to measure biochemical reactions that occur on the surface of the microspheres.
  • The third is a high-speed digital signal processor to efficiently manage the fluorescent output.
Applications
  • Multiplex analysis of cytokines and phoshoproteins with prepared kits available from BioRad
  • Multiplex analysis of a variety of proteins by conjugating antibodies to beads via kits available from BioRad

Location —The BioPlex system is located in the Building 1 Room B60 lab of the IRF.
IRF Contact: Udai Singh
Websitewww.Biorad.com/

Kodak Image Station 2000MM

Description — The Kodak ISM 2000MM is a fluorescence based molecular imaging station equipped with long wavelength full field illumination. Depending on available filters placed in the system green to near IR excitation and emission can be used for imaging luminescent, fluorescent, radiographic and colorimetric labeled biomolecules in a variety of formats including membranes, electrophoresis gels, plates and tissues.

  • Available excitation filters are for wavelengths of 465nm, 535nm and 620nm. Corresponding emission filters are for 533nm, 600nm and 700nm.
  • Kodak 1 D Image Analysis Software
Applications — Documentation and analysis of:

  • DNA, RNA and protein electrophoresis gels
  • Southern, Northern and Western blots
  • Dot and Slot blots
  • Microplate assays
  • Macro Arrays
  • Automated cell colony counting
  • Plaque screening
  • Thin layer chromatography

Location - The IS2000MM is located in Building 1 Room A10
IRF Contact: Jay Potts
Website - www.jaewootr.com/kodak/IS2000MM/is2000mm.ht

Agilent 2100 Bioanalyzer

Description — The Agilent 2100 Bioanalyzer is a microfluidics based platform for the analysis of DNA, RNA, proteins and cells. The lab-on-a-chip based system is designed for rapid, automated, digital acquisition and analysis of data.

Applications

  • Optimization of PCR reactions for gene expression, sequencing, cloning and typing
  • Quality control analysis for ribosomal RNA contamination in mRNA prior to microarray analysis
  • DNA and RNA LabChip kits for quality control of probes and targets
  • Analysis of RNA integrity
  • Protein 200 Plus Lab Chip assay for rapidly (30 minutes or less) quantifying and sizing protein samples
  • Protein analysis in cell lysate samples
  • Analysis of protein purit
  • Quality control for analysis of antibodies

Location- The Agilent 2100 Bioanalyzer is located in the Building 1 Room B60 laboratory of the IRF’
IRF Contact:  Jay Potts

BioRad iCycler iQ and Applied Biosystems Step One Plus Real Time PCR Systems

Description — Real time PCR, in conjunction with the appropriate fluorescent detection strategies, allows the initial amount of nucleic acid in a reaction to be determined. Advances in the technology have led to significant improvements in the accuracy of DNA quantitation and gene expression analysis. Quantitation is achieved by measuring the increase in fluorescence during the exponential phase of the PCR reaction. The system in the IRF is equipped for:

  • Three color analysis for amplification of three dependant targets
  • Taqman, Sybr Green, primer based system and molecular beacon analyses
  • Multiple sample block formats for 0.2ml tubes, 96 well plates and 384 well PCR plates
  • Real time display of data acquisition
Applications
  • Quantitative comparison of genes of interest in housekeeping genes in a single sample
  • Quantitative detection of gene expression in genetically modified organisms
  • Analysis of single nucleotide polymorphisms (SNPs)
  • When used in conjunction with laser capture microdissection, real time PCR provides an excellent technique for comparison of genetic expression in specific cell types such as cancer vs. normal cells from a single region of tissue.

Location:    The BioRad iCycler iQ and ABI StepOne Plus Real Time PCR Systems are located in the Building 1 Room B60 laboratory of the IRF

IRF Contact: Jay Potts

Websites – www.BioRad.com/ and www.amebioscience.com/pcr.htm

 

BioRad Molecular Imager FX

Description — The FX is an isotopic and fluorescent imaging system capable of collecting and analyzing radioisotope emission, chemiluminescence and fluorescence data

  • Dectection of 32P, 33P, 35S, 14C, 3H, and 125I, HRP-activated chemiluminescent substrates and fluorescence using phosphor screen technology
  • Dual wavelenght 532 and 1064 internal laser
  • External 488 and 635 lasers
  • Scan resolutions of 800, 200, 100 and 50 microns
  • Quantity One Software and PDQuest software packages
Applications

  • Scans phosphor screens, gels, blots, microtiter plates, high density arrays
  • Imaging and quantitation of a broad range of sample types over a large dynamic range
  • Location- The FX is located in Building 1 Room B51
  • Identification of spots in 2D gels
  • DIGE – Fluorescent image acquisition for Differential Gel Electrophoresis

Location – The FX is located in Building 1 Room A2

IRF Contact: Jay Potts

Website - www.BioRad.com/

Websitewww.BioRad.com

BD FACS Aria II

Description – The FACS-Aria II high performance cell sorter is a flow cytometer capable of sorting upto 70,000 events per second while maintaining viability and function of cells in culture. This makes it an ideal system for sterile cell sorting for separating rare cell events, such as stem cells, dendritic cells, and NKT cells that often represents less than 1% to 2% of the total cells in a large population.

  • Solid state 488 and 640 lasers and sorting capabilities.
  •  Automated Cell Deposition Unit and Aerosol Management system.
  • The 488 line has forward and side scatter and 4 fluorescent detectors. The 640 line has 2 fluorescent detectors.
  • Two and Four way sorting into 12X75mm tubes, 15 ml tubes, slides or culture plates.
  • Fluidics with 70, 85 and 100 micron nozzle tips.
  • Temperature control option for cooling of sort collection devices – tubes and plates
  • Optical filters for the 488 laser include FITC, PE, PE-Texas Red, PerCP, CY5.5 and PECY7.
  • Optical Filters for the 640 laser includes APC-CY7, Alexa700, APC
  • Computer and FACSDIVA software
  • Printer

Applications:

  • Analysis of cell surface antigens and biomarkers
  • Immunophenotyping
  • Cell size and granularity
  • Stem cells and hematopoietic cell lineage studies
  • Hematology
  • Apoptosis, live/dead cell assay
  • Cell cycle studies
  • Stem cell 4 way sorting
  • Cell viability and Functional assays
  • Rare cell population sorting
  • Slides and culture plate sorting

Location – The FACS Aria II is located in Building 1 Room B44 of the IRF

IRF Contact: Udai Singh

Website – http://www.dakocytomation.us

Miltenyi autoMACS Pro Separator

Description – A benchtop automated magnetic cell sorter for the isolation of virtually any cells type from any species based on the Miltenyi MACS technology. Cells are sorted by use of 50nm magnetic beads with attached antibodies

  • Automated processing of  up to 6 samples at a time
  • Sample volumes of 0.2 to 50ml
  • Separates up to 107 cells/second

Applications

  • Separation of rare cells such as CD 34+ and CD133+ stem and progenitor cells
  • Separation of antigen speficic T-cells
  • Separation of dendritic cells and their subtypes
  • Separation of frequent cells such as CD4+ T helper cells, CD8+ cytotoxic T cells, monocytes and B cells
  • Cell sorting directly from whole blood volumes of up to 15ml
  • Can be used as a pre-sorting technique for increased speed in flow cytometric sorting

Website – www.miltenyibiotec.com/en/PG_651_679_autoMACS_Pro_Starting_Kit.aspx

IRF Contact: Udai Singh

Location – The Miltenyi autoMACS Pro is located in the Building 1 Basement lab of the IRF

Beckman Coulter FC500 Flow Cytometer

Description – The Beckman Coulter FC-500 is a flow cytometer designed for qualitative and quantitative measurements of biological and physical properties of cells and fluorescent particles. This system is equipped with:

  • Argon ion 488 nm 20mW output laser
  • Red Helium-neon 633 nm 20mW output laser
  • Optical filters for 5 color analysis
  • Flow rates up to maximum 3300 events per second
  • 12X75 glass or plastic tubes
  • Single tube sampling mode
  • 32 tube Multi Carousel loader (designed for vortexing each tube individually).
  • Computer and CXP software
  • Printer

Applications:

  • Analysis of cell surface antigens and biomarkers
  • Immunophenotyping
  • Cell size and granularity
  • Stem cell and hematopoietic cell lineage studies
  • Hematology
  • Apoptosis, live/dead cell assay
  • Cell cycle studies
  • Cell viability and Functional assays
  • DNA analysis

Location – The FC 500  is located in Building 1 Room B44 of the IRF

IRF Contact: Udai Singh

Website – http://www.beckmancoulter.com/

Sysmex KX21N Hematology Analyzer

Description - the Sysmex KX-21N is ideal for near patient, clinic or satellite lab testing of hematology samples

  • Offers 19 parameters including a 3-part WBC differential plus histograms for RBC, PLT and WBC counts
  • Whole Blood Mode (50ml sample) – WBC, RBC, HGB, HCT, MCV, MCH, MCHC, PLT, LYM%, MXD%, NEUT%, LYM#, MXD#, NEUT#, RDW-SD, RDW-CV, PDW, MPV, P-LCR
  • Predilute Mode (20ml sample) - WBC, RBC, PLT, HGB, HCT, MCV, MCH, MCHC
  • Completely automatic START-UP function, one button sampling and single button daily maintenance
  • Six QC files on-board – Results are automatically entered into the L-J or X bar chart for comprehensive daily quality control

Location: The Sysmex KX21N system is located in the Building 1 Basement lab of the IRF

IRF Contact: Udai Singh

Websitehttp://www.labsco.com/intranet/uploads/1kx-21n_flyer.pdf

Zeiss PALM CombiSystem for Laser Capture Microdissection with Optical Tweezers

Description – The PALM laser capture microdissection system is designed for the isolation of specific cell types or regions of tissue from sections of tissues processed by paraffin embedding or cryo-sectioning. This allows for the molecular analysis and comparison of pure populations of cells or specific regions of tissues. The optical tweezers allows movement and positioning of specific cells from cell cultures. The IRF PALM system is equipped with:

  • Zeiss Axiovert 200 inverted microscope platform
  • A range of objectives used for fluorescence imaging and cutting
  • Nitrogen 337nm and Infrared lasers
  • Red and green fluorescence capabilities

Applications

  • Isolation and removal of pure cell populations from complex tissues such as cancer cells from the surrounding stromal tissue or normal cells within a tumor
  • Isolation of fluorescently labeled cells such as cells transfected with GFP from the surrounding population of cells and tissue
  • Capture of live cells for further culture with use of Pen Membrane frames
  • Analysis of DNA, RNA and proteins from paraffin embedded and cryo-sectioned material

Location: The PALM system is located in Bldg 1 Room B54 of the IRF

IRF Contacts: Bob Price or Anna Harper

Website – http://www.zeiss.com/palm

Beckman Coulter DU800 UV/Visible Spectrophotometer

Description – The DU 800 The DU 800 Spectrophotometer is a PC controlled system intended for use in quantitative and qualitative analysis in biological and industrial procedures that require spectrophotometric measurements in the UV and visible region of the electromagnetic spectrum.

  • Detection from 190-1100nm wavelengths in !1.8nm bandwidths
  • Equipped for small volume samples
  • Detects absorbance and transmittance

Applications

  • Useful for kinetics and time measurements, nucleic acid, protein and enzyme analyses

Location – The DU 800 is located in Building 1 Room A2

IRF Contact: Udai Singh

Website - www.BeckmanCoulter.com/