Imaging Facility


General Information

BCIF Facility 1
BCIF Facility 2

Biological Chemistry Imaging Facility (BCIF) provides resources for data acquisition and analysis for radioactive, fluorescent, and photographic samples as well as digital imaging and document production. BCIF started operations in December 1994 as a small departmental facility. Over the years it grew into a core imaging facility serving the majority of departments and subdivisions of the UCLA David Geffen School of Medicine.

BCIF provides round-the-clock access to a cluster of modern equipment such as Typhoons 9410 and 9400 Variable Mode Imagers, Storm 820 and 840 imagers, Laser Densitometer, Gel Documentation System, color printers, and high-resolution scanners. In addition BCIF provides unlimited scientific data storage space on its secure storage arrays to all participating labs, as well as access to a number of software packages for data analysis and digital data processing.

All equipment in the facility is available 24 hours a day, 7 days a week. The facility operates on a walk-in basis and provides technical support and instructions during weekdays, primarily for new users. Each lab is provided with its own account and space on a file server. Any lab wishing to establish a new account is welcome to do so by contacting either Imaging Lab Director Dr. Michael Haykinson (mhaykinson@mednet.ucla.edu or x50786) or Lab Manager Ric Grambo (BCIF@mednet.ucla.edu or x61415).

BCIF Locations
BCIF has two locations: 344C BSRB for labs located at the Bioscience Research Building and 33-089 CHS for labs located at other campus buildings.

Fee Structure
An annual fee of $600 per lab is required for use of the BCIF facility. Please note that the number of users per lab does not affect the charge. This fee also covers black/white and color laser printing on regular paper. Printing on specialty paper or transparencies is charged separately.

Equipment

Typhoon™ 9410 and 9400 Variable Mode Imagers from GE Healthcare are the main equipment pieces at both BCIF locations. Typhoon imagers are versatile systems, which handle agarose and polyacrylamide gels, membranes, mounted or unmounted storage phosphor screens up to 35×43cm, microplates, microarrays, and in situ slides.

Typhoon Machinestorage phosphor

  • direct blue-excited fluorescence (457 nm, 488 nm)
  • direct green-excited fluorescence (532 nm)
  • direct red-excited fluorescence (633 nm)
  • chemiluminescence

 

 

Important Typhoon features:

  • Unique Typhoon property: Highly sensitive confocal optics enables direct and highly quantitative imaging of fluorescent Western blots without intermediate exposure to films or screens
  • Powerful excitation sources and confocal optics allow for the sensitive detection of very low-abundance targets
  • Red-, green-, and two blue-excitation wavelengths and a wide choice of emission filters enable imaging of a variety of fluorophores across visible spectrum
  • Automated four-color fluorescence scanning allows multiplexing of multiple targets in the same sample (i.e. Cy2, Cy3, and Cy5)
  • Storage phosphor autoradiography delivers high-resolution imaging and accurate quantitation of 3H,14C,125I, 32P, 33P, 35S, and other sources of ionizing radiation
  • Typhoon 9410 imager can image microarrays and in situ slides at 10 micron resolution
  • The Typhoon exhibits outstanding linearity (4-5 orders of magnitude), quantitative accuracy, and extremely low limits of detection.

TECHNICAL SPECIFICATIONS

Scan area
35 × 43 cm
Sensitivity (limit of detection) 
Storage Phosphor: 14C (1 h exposure, 200 and 100 µm only): <2 dpm/mm2;32P: 5-10-fold lower than 14C
488 nm excited fluorescence: 100 amol FAM end-labeled DNA primer in 12% polyacrylamide gel sandwich, 0.4 mm thick
532 nm excited fluorescence: 200 amol HEX, TMR, ROX, and 400 amol FAM end-labeled DNA primer in 12% polyacrylamide gel sandwich, 0.4 mm thick
633 nm excited fluorescence: 200 amol Cy5 end-labeled DNA primer in 0.4 mm thick, 12% polyacrylamide gel with TBE buffer.
Pixel size
1000, 500, 200, 100, 50, 25 (9400 model), and 10 µm (9410 model), selectable
Spatial resolution
Autoradiography:2 line pairs/mm for lines drawn with14C ink.
Blue-excited fluorescence: 10 line pairs/mm
Green-excited fluorescence: 10 line pairs/mm
Red-excited fluorescence: 10 line pairs/mm
Uniformity
± 5% over entire scan area.
Pixel accuracy
± 0.15%
Data format
16-bit (65,536 levels) TIFF
Linear dynamic range and linearity
Five orders of magnitude (100,000:1) with less than 7.5% relative standard deviation for entire dynamic range.
Red light source
Type: 10 mW Helium-Neon laser Estimated average lifetime: ~ 10,000 h Wavelength: 632.8 nm
Green light source
Type: 20 mW solid-state doubled frequency SYAG laser  Estimated average lifetime: ~10.000 h Wavelength: 532 nm
Blue light source
Type: 30 mW all blue lines Argon ion laser  Estimated average lifetime: ~5,000 h Wavelength: 488 nm (20 mW) and 457 nm (4 mW)
External interface
10 Base-T Ethernet using TCP/IP protocol
Software
Scan control software for Windows™ operating system.
Power requirements
115/230 V (switchable), 50-60 Hz, < 500 W
Dimensions
(W × D × H) 
Instrument: 118 × 78 × 48 cm
Blue laser module: 30 × 78 × 48 cm

Microarray Slide


A microarray slide, spotted with cDNA and hybridized with 25 pmol of Cy3-labeled skeletal muscle mRNA and Cy5-labeled liver mRNA. The slide was scanned on Typhoon 9410 at 10 µm pixel size resolution.

 

2D Protein Gel

 

 

Two-dimensional 2-D protein gel stained with Deep Purple™ Stain. Scanning: Typhoon 9400: Excitation, green laser (532 nm); emission 560LP filter, PMT 530 V, 100 µm resolution. The gel image shows the pH 3-8 region, where most proteins are present.

 

Four-color competitive gel shift assayFour-color competitive gel shift assay to determine the relative affinities of a protein binding td four different oligonucloetides labeled with FAM, HEX, TAMRA and ROX respectively at the same time. The intensities
for each fluorophore in a lane are related directly to the relative affinity of the repressor protein for the oligonucleotide used.

updated October 17, 2013

2009 Department of Biological Chemistry. All Rights Reserved.