Typhoon imaging system and the 2D gel documentation/analysis system
The facility has two methods for the acquisition of digital gel
images. For the acquisition of Coomassie Blue stained gels, the
Amersham Pharmacia Biotech
ImageScanner flatbed scanner with transparency module can be used.
Once the gel image has been digitized, 2-D gel analysis can be
performed using Progenesis SameSpots from Nonlinear Dynamics.
SameSpots allows for the alignment and analysis of 2D gels in a short amount of time and
returns data that shows the expression of proteins under different biological
conditions. The software presents a list of the most significantly changing spots within a set of
gels, allowing for the rapid screening of protein spots that may be of the most interest in a set
of gels. These spots can then be further analyzed by other equipment in the Protein Facility
(trypsin digestion followed by peptide mass fingerprinting or internal sequencing, direct N-
terminal sequencing of the blotted gels). SameSpots uses pixel-level alignment that allows for 100% spot matching.
This eliminates missing values and reduces the need for spot-editing. Each spot on a set of gels will use the same spot outline making comparisons of replicate gels under different conditions much easier and more accurate.
An older software package (PDQuest from Bio-Rad) is also available.
The Typhoon 8600 from Amersham
Pharmacia Biotech is a multimode image scanner that can be used
for the acquisition of phosphorescence, fluorescence or
chemiluminescent samples. The scanner is capable of imaging gel
sandwiches, agarose and polyacrylamide gels, membranes, microplates
and microarrays. Samples can also be labeled with multiple dyes and
imaged on the Typhoon. After image acquisition, the image can be
analyzed for quantitative data using ImageQuant software from
Amersham Pharmacia Biotech.
