We use a variety of approaches in our research

Goat blastocyst stained for nucleolin (red) and NANOG (green).

Goat blastocyst

Stained for TRA-1-60

Goat embryo stained for catenin.

Cat seminiferous tubule stained for vimentin (red) and DNA (blue).

Immunocytochemistry

Immunocytochemistry (ICC) and immunohistochemistry (IHC) provide a means of visualizing the expression and location of protein within cells and tissues, respectively. ICC generally involves fixing cells or embryos prior to immunostaining, while IHC involves multiple steps including fixation, embedding, and sectioning the processed tissues.

mESC expressing GFP driven by bNanog promoter

Mouse blastocyst expressing GFP (DNA stained blue)

Incorporation of reporter transgene into mESC chromosome (FISH)

Transgenic Reporters

Genes encoding fluorescent proteins can be linked to promoters and other regulatory elements which control the expression of the fluorescent protein. These can be constitutive promoters such that the cells continually express the reporter protein, which is useful for tracking the cells. Alternatively, the reporter gene can be used to study the regulation of a particular promoter or to monitor a particular cell phenotype.

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Western blot stained for transcription factors NANOG and OCT4.

GC-MS profile of epididymal sperm and fluid (domestic cat)

Molecular Techniques

Fluorescent In Situ Hybridization (FISH), Western blots, RT-PCR & RT-qPCR

Cellular Techniques

Embryo and cell culture, embryo microinjection, somatic cell nuclear transfer (SCNT), & cell fusion

Immuno-based Techniques

ICC, IHC (see above), immunoprecipitation, cell separation (MACS/FACS)

Gas Chromatography-Mass Spectrometry (GC-MS)

GC-MS in collaboration with the Sunny laboratory (ANSC) and the Sriram laboratory (ChBE)

Atomic Force Microscopy (AFM)

AFM in collaboration with the Desai laboratory (MECH)