CHAPTER 3
20
Operation
Microplates that meet certain physical characteristics can be
scanned directly on the Odyssey
®
scanning surface. Proper selection
of microplates significantly affects the results of your analysis as each
plate has its own characteristics including well depth, plate autoflu-
orescence, and well-to-well signal crossover. Some general consid-
erations for microplate selection are provided below.
•
Plate dimensions must be such that the distance from the Odyssey
scanning surface to the target detection area of the plate is 4.0 mm or less.
•
In order to avoid well-to-well signal spread, black-walled, clear bottom
plates should be used for assays that involve imaging of a liquid. Since
In-Cell Western
™
assays use detection at the well surface with no liquid
present, both clear and black-walled plates can be used. Consult protocols
in the Application Protocols manual or on the LI-COR
®
website
(
http://biosupport.licor.com
) for specific recommendations.
•
Do not use plates with white walls because the autofluorescence from the
white surface will create significant noise.
•
For In-Cell Western assays requiring sterile plates for tissue culture growth,
the following plates are recommended by LI-COR Biosciences.
96-well format Nunc
®
(P/N 161093) Clear
96-well format Nunc (P/N 165305) Black
96-well format Falcon
™
(P/N 353075) Clear
96-well format Falcon (P/N 353948) Black
384-well format Nunc (P/N 164688) Clear
384-well format Nunc (P/N 164730) Black
384-well format Falcon (P/N 353961) Clear
384-well format Falcon (P/N 353962) Black
•
Before plate scanning, clean the bottom plate surface with a moist, lint-
free paper to remove any obstructions. Additionally, the Odyssey scanning
surface should be thoroughly cleaned using the procedures described
earlier in this chapter.
Using Microplates
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