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Rocky Mountain Neurological Disorders Center Core

Microscopy Core

Mark Dell'Acqua, Director

Advanced Light Microscopy Core provides advanced light microscopy services including 2-photon, SHG (second harmonics generation), lifetime imaging (FLIM) modalities, CARS (Coherent anti-Stokes Raman spectroscopy), fluorescence correlation techniques (FCS, RICS), high resolution laser scanning confocal and fast spinning disk confocal with FRAP (fluorescence recovery after photobleaching) and FRET (fluorescence resonance energy transfer) capabilities, TIRF (total internal reflection fluorescence) as well as super-resolution imaging (i.e. STED - stimulated emission depletion, PALM - photoactivated localization microscopy, STORM - stochastic optical reconstruction microscopy). (see for the full list of instruments). Individuals who want to use the light microscopy facility should contact Radu Moldovan.


Targeting Vector Core

The core specializes in generating gene targeting and transgenic mouse models.  The core has been serving the University of Colorado community since 2005.  In that time, we have made over seventy vectors including knock-in, knock-out, conditional knock-out, traditional transgenic, and BAC transgenic.  We have also developed specialized vector models for knock-in at the ROSA26 locus.  For gene targeting models, our success rate for correct ES cell targeting is 95% and for germline transmission is 100%.   The core also offers additional support services such as ES cell screening, BAC purification and mouse genotyping.  

For new project requests, please contact Wallace Chick.

Wendy Macklin - Director
Wallace Chick - Co-Director

Vector Types

Conditional Knock-out  
This vector achieves conditional knock-out of your GOI by deleting a pertinent exon.  The exon is floxed and can be deleted by introducing Cre to the system.  The vector contains a Neomycin positive selection marker and a TK negative selection marker. 


This vector is useful for introducing a mutation to the genome.  The mutation could be a base pair change, an amino acid change or fusing the expression of your gene to a marker (ex. Cre).  The vector contains a Neomycin positive selection marker and a TK negative selection marker.


Knock-in at the ROSA26 Locus by RMCE 
This model is useful for either constitutive or conditional expression of your cDNA at the ROSA26 locus.  This method is fast, efficient and results in a more consistent expression of cDNA as compared to traditional transgenic models.  Your cDNA can be cloned into one of our pre-designed exchange vectors.  The exchange vector is then electroporated into ES cells which already contain the tagged ROSA26 allele.  Efficient exchange between the FRT and F3 sited results in incorporation of your cDNA into the ROSA26 locus. 

Flex Switch
This model achieves a simultaneous deletion of an exon and activation of a marker by utilizing the Flex Switch technique described by Schnutgen et. al. FLEx switch.pdf  
Initially, the GOI functions normally and no marker is expressed.  By utilizing specific recombination sites, loxP and lox511, deletion of exon and activation of marker occurs simultaneously. The expression of the marker can be driven by the endogenous promoter or a strong exogenous promoter (ex. CAG).



BAC Transgenic
The expression of a marker is driven by the promoter of GOI.  The use of BACs has several advantages 
including the presence of regulatory elements and minimal positional effects.  The marker can be of 
several types including fluorescent tags and Cre derivatives.   


For clients outside of the University of Colorado, add another 10% to the prices.
Prices are valid for the 2017 fiscal year
Transgenic or RMCE Vector:                          $3,385.00
KI/KO, BAC Transgenic Vector:                      $3,826.00
Flex Switch Vector:                                       $4,708.00
ES Cell Targeting:                                          $2,967.06
ES Cell Screening (per 96-well plate):               $685.00
BAC purification:                                              $169.12 
Gel Purification:                                                $121.97
Genomic DNA isolation (ES cells):                     $229.00 set up, $12 per clone
Long Range PCR:                                             $114.00 set up, $12 per sample
Mouse Genotyping (per sample):                       $12.50 (min. $50)

Zebrafish Transgenic Core

Angie Ribera, Director

The NINDS P30 Center Zebrafish Core has three Specific Aims:

  1. Create transgenic lines of zebrafish that express different fluorescent proteins in specific populations of pre- and postsynaptic neurons
  2. Create transgenic zebrafish strains that express genetically-encoded calcium indicator dyes in specific neuronal populations
  3. Maintain transgenic and wild type zebrafish strains for the UCD neuroscience community

During the first year of the grant, the fish facility at the Fitzsimons campus is being optimized and we are focusing on Aim 3. In subsequent years, Aims 1 and 2 will be a priority. The Core can cover the expenses associated with breeding wild type fish for pilot experiments. If you would like to use zebrafish embryos for pilot experiments, please contact Angie Ribera for further information.

Zebrafish transgenic lines express GFP in specific neuronal subpopulationsZebrafish transgenic lines express GFP in specific neuronal subpopulations. In the Figure, GFP expression is driven by the flh promoter. (This transgenic line was developed by Drs. Marnie Halpern and Josh Gamse - Carnegie Institute of Washington). At 48 hours post fertilization, GFP expression (green) is present in specific populations of motor neurons that extend axons either ventrally (down, asterisks) or dorsally (up, arrowhead). A subset of the axons also expresses the zn-8 epitope (red).  Zn-8 is a marker for axons of later-born (secondary) motor neurons.
Scale Bar: 25 µm.

Machine Shop

Michael Hall, Core Director

The Neuroscience core machine shop is located in room NG003 on the ground floor of Building 500 (northwest corner) at the Anschutz Medical Campus (View Map). The shop's equipment and staff are there to meet the custom design and machining needs of all the faculty, staff and students of the UC Denver-Anschutz Medical Campus.


Click to view larger image of LAGUN 


The shop machinery includes:

    • 1 manual milling machine with digital readout
    • 1 CNC milling machine (full 3-axis automation)
    • 2 Lathes (one high-precision)
    • 3 wood/metal-cutting band saws
    • 1 table saw
    • 1 precision surface grinder
    • 1 drill press
    • 1 12" disk/6" belt sander
    • 1 52" shear
    • Extensive tooling & precision measuring devices

Click to view larger image of SHELDONThe shop routinely stocks a modest amount of material (acrylic, aluminum, brass, hardware) for small projects.  The shop labor rate is $68/hour for members of the Center for Neuroscience and $92/hr for all others. Charges are based upon project time and material costs. Payment for services is made with a current university speedtype number or can be billed to outside sources.


The shop is staffed by one machinist 24 hours/week although there are currently no routine hours of operation. Contact Michael Hall at 303-724-1335 or, preferably, by e-mail at to discuss a project, to arrange a meeting or with questions.