Cellular Imaging Core

About Us

The Cellular Imaging Core provides state-of-the-art equipment in microscopy and digital imaging to investigators at CHLA. Imaging technology in modern biological research is constantly undergoing enormous advances. The acquisition, processing, analysis and storage of digital images are an important part of most biomedical research projects. The primary focus of the Core is technology transfer and provision of facilities for acquisition and analysis of histological and cytological preparations. An important aspect of this technology transfer is to provide training in the use of digital imaging devices, as well as in the application of image analysis procedures for generation of quantitative data.

Services

The Imaging Scientist provides expert consultation on the design, execution and analysis of fixed and live cell imaging experiments. Please contact the Imaging Scientist directly to arrange access and training in the utilization of Core facilities.

This Core participates in the CHLA Core Pilot Program. To learn more click here.

CHLA welcomes external users to utilize our Core facilities. Please contact G. Esteban Fernandez, PhD at GEFernandez@chla.usc.edu or call 323-361-2548.

  • Equipment and Software
  • Fee Structure
  • Published Data

The Cellular Imaging Core houses modern light microscopy and digital image processing equipment to support cell biology research at our facility as well as other institutions.

All microscopes are equipped with high-resolution objective lenses and digital cameras to produce the highest quality images and time-lapse movies.

Zeiss Axio Observer 7 fluorescence live cell imaging microscope (Saban 542)

The Zeiss Axio Observer 7 live cell imaging microscope is capable of imaging live specimens using widefield fluorescence, brightfield, phase-contrast, and/or DIC/Nomarski with an sCMOS camera for speed, sensitivity, and a large field of view at high resolution. It is equipped with a robust focus stabilization system and an environmental chamber for stable long-term imaging. A programmable motorized stage allows for automated imaging of multiple positions in parallel, for example of different treatment groups in a multi-well chamber. The objective lenses on this microscope range from 5x to water-immersion 63x. It has an LED light source for gentle illumination and extremely fast switching between colors. An ApoTome structured illumination system is available for optical sectioning to remove out-of-focus haze. The fluorescence excitation LEDs available are:

  • 385 nm for UV-excitable dyes such as DAPI
  • 425 nm for CFP
  • 469 nm for green dyes such as Alexa Fluor 488, FITC, and GFP
  • 511 nm for YFP
  • 555 nm for red dyes such as Alexa Fluor 546, Cy3, rhodamine, RFP, and tdTomato
  • 590 nm for longer red dyes such as Alexa Fluor 568 and 594 and mCherry
  • 631 nm for far-red dyes such as Alexa Fluor 633 and 647, Cy5 and TO-PRO-3

LaVision Biotec UltraMicroscope II fluorescence lightsheet microscope (Saban 538)

Lightsheet microscopy is capable of imaging large specimens, such as whole mouse embryos and organs, in 3D rapidly and at high resolution. In this technique, a thin sheet of light optically slices through a cleared specimen. A 3D image is built by moving the sheet through the depth of the specimen while a synchronized camera captures an image of each optical section. Utilization of sCMOS camera technology, with its high sensitivity and fast data transfer rate, allows for short exposure times and much faster acquisition times than point-scanning confocal. Since illumination is orthogonal to the direction of image capture, each plane is illuminated only once and the specimen suffers substantially less photobleaching. Additionally, the UltraMicroscope II is unique in its ability to image specimens cleared with organic solvents for superior clarity over aqueous techniques.  Superb specimen clarity combined with minimal photobleaching and a high numerical aperture long working distance lens allow the UltraMicroscope to generate high-resolution images at increased depth, up to about 6 mm; it can accomodate specimens up to 1.2 x 1.2 x 0.6 cm. It is equipped with a zoom body to easily change between acquiring fast broad-view images and achieving single-cell resolution (down to 0.5 microns/pixel). The fluorescence excitation lasers available are:

  • 405 nm for blue dyes such as DAPI and CFP
  • 488 nm for green dyes such as Alexa Fluor 488, FITC, GFP, and YFP
  • 561 nm for red dyes such as Alexa Fluors 546 and 568, and 594, Cy3, rhodamine, RFP, tdTomato, and mCherry
  • 640 nm for far-red dyes such as Alexa Fluors 633 and 647, Cy5 and TO-PRO-3

UltraMicroscopeII Lightsheet Microscope A 75%.jpgUntitled.jpg

Zeiss LSM 710 Inverted Confocal Microscope (Saban 338)

  • Specializes in three-dimensional fluorescence imaging
  • Acquisition of up to 8 fluorescence channels simultaneously
  • Motorized stage for tilting/stitching large areas
  • DIC/Nomarski imaging simultaneous with fluorescence
  • Capable of advanced analytical techniques like FRAP, FRET and spectral unmixing

The fluorescence excitation lasers available are:

  • 405 nm for blue dyes such as DAPI
  • 488 nm for green dyes such as Alex Fluor 488, FITC, and GFP
  • 555 nm for red dyes such as Alexa Fluor 546, 568 and 594, Cy3, tdTomato, and mCherry
  • 639 for far-red dyes such as Alexa Fluor 633 and 647, Cy5 and TO-PRO-3

Zeiss-LSM-710 50%.jpg

LSM 700 Inverted Confocal Microscope (Saban 448)

  • Specializes in confocal and three-dimensional flourescence imaging
  • Acquistion two fluorescence channels simultaneously
  • Capable of advanced analytical techniques like FRAP, FRET and spectral unmixing
  • DIC/Nomarski imaging simultaneous with fluorescence
  • Motorized stage for tiling/stitching large areas

Leica DMI6000B Inverted Microscope (Smith Tower 1030B)

  • Versatile microscope that can image virtually any specimen on a slide or culture dish/plate
  • DIC/Nomarski and phase-contrast optics
  • Color CCD camera for histology and immunohistochemistry slides
  • Fast, ultra-sensitive high-resolution sCMOS camera for epifluroescence
  • Motorized stage for tiling/stitching large areas

Fluorescence filters for:

  • UV-excited dyes such as DAPI
  • Green dyes such as Alex Fluor 488, FITC, and GFP
  • Red dyes such as Alexa Fluor 546, 568 and 594, Cy3, tdTomato, and mCherry
  • Far-red dyes such as Alexa Fluor 633 and 647, Cy5 and TO-PRO-3

Leica MCFLIII Fluorescence Stereoscope & Optical Projection Tomography (OPT) Microscope (Saban 439)

  • Specializes in 3D imaging of organs, embryos and other whole specimens
  • Filters for UV, green, red, and far red fluorescent dyes and protein
  • Capable of reflected and transmitted light
  • Motorized focus (z) drive for “extended depth of focus” images
  • Capable of tomographic 3D imaging (OPT) of fluorescent and colorimetric labeling

Leica_MZFLIII_Stereo_Microscope 50%.jpg

Zeiss Axioplan Upright Microscope (Smith Tower 1030B)

  • Specializes in color brightfield imaging
  • Color CCD camera

Zeiss_Axioplan_Upright_Microscope 50%.jpgColor_Brightfield - Zeiss Axioplan Upright Microscope 50%.jpg

CARV II Spinning Disk Live Cell Imaging Confocal (Smith Tower 1030B)

  • Nipkow spinning disk confocal system with lamp-based excitation (no lasers) for gentler confocal imaging of live cells
  • Enclosed in environmental chamber to control temperature, CO2, and O2 for imaging live cells over extended time
  • Capable of standard blue, green, red and far red fluorescence in confocal or widefield mode
  • Optics for DIC/Nomarski and phase contrast imaging

Image Processing Workstations and Software 
(Saban 543 and Smith Tower 322C)

  • Arivis Vision4D - for visualization and analysis of static and timelapse 3D images, including cell/particle tracking. Specialized for efficiently processing very large datasets, such as lightsheet images. 
  • MetaMorph – powerful 2D and 3D image processing and analysis including morphometry and movie making
  • AutoDeblur – very user-friendly automated deconvolution of widefield, confocal, and lightsheet data to reduce blurring and noise and increase resolution
  • Photoshop – image preparation for publication and presentation
  • FoveaPro – scientific image processing and analysis add-ons for Photoshop
  • ImageJ – versatile, customizable, open-source scientific image processing and analysis software.  The core’s Imaging Scientist can write custom software for ImageJ to suit your needs
  • MATLAB Image Processing Toolbox - fast, powerful programming environment for image processing

Cellular Imaging Core Fee Structure

Pricing Table.jpg

1= 2D imaging techniques available: fluorescence; color brightfield, phase-contrast & differential interference contrast (DIC); automated tiling & merging of large areas 

2 OPT= optical projection tomography 

Access for Non-Hospital Researchers

Access to Cellular Imaging Core facilities for researchers at USC and other institutions can be arranged. Interested researchers should contact the Imaging Scientist to inquire about availability and rates.

How to cite the Core in publications:

The authors thank G. Esteban Fernandez and the Cellular Imaging Core at the Saban Research Institute of Children's Hospital Los Angeles for expert assistance with image acquisition and/or analysis. 

Cellular Imaging Core Publications and Data 

3D Confocal

Vimentin-mediated signaling is required for IbeA+ E. coli K1 invasion of human brain microvascular endothelial cells.
2010. Chi F, Jong TD, Wang L, Ouyang Y, Wu C, Li W, Huang SH. Biochemical Journal 427(1):79-90. 

Calcium Imaging In Live Cells 

Escherichia coli K-1 Interaction with Human Brain Micro-vascular Endothelial Cells Triggers Phospholipase C-γ1 Activation Downstream of Phosphatidylinositol 3-Kinase
2003. Sukumaran SK, McNamara G, Prasadarao NV. Journal of Biological Chemistry 278(46):45753-45762. 

Confocal Image Quantitation

Hypothalamic neural projections are permanently disrupted in diet-induced obese rats.
2008. Bouret SG, Gorski JN, Patterson CM, Chen S, Levin BE, Simerly RB. Cell Metabolism 7(2):179-85.   

Fluorescence Image Quantitation

The Contribution of Bone Marrow–Derived Cells to the Tumor Vasculature in Neuroblastoma Is Matrix Metalloproteinase-9 Dependent
2005. Jodele S, Chantrain CF, Blavier L, Lutzko C, Crooks GM, Shimada H, Coussens LM, DeClerck YA. Cancer Research 65(8):3200-3208. 

Spectral Imaging

Hepatitis C Virus Causes Uncoupling of Mitotic Checkpoint and Chromosomal Polyploidy through the Rb Pathway
2009.  Machida K, Liu J-C, McNamara G, Levine A, Duan L, Lai MMC. Journal Of Virology 83(23):12590–12600.