Mitsubishi P91DW User Manual download pdf (Page 57)

Alpha Innotech User’s Manual, version 4

3-12

Flat Field Calibrate

Flat Field Calibrate is a function that is used to ‘flatten’ the image so that the pixel data

is even across the entire image area. This is a function that is useful for large gels and

other applications that use the entire field of view for an image.

Creating flats can be art in itself; there are many documents on the internet that can help

users interested in this arena to create the ideal flat for the application. However, some

useful flats that have been created in the past involve very simple tools like a piece of 8.5

x 11 regular low quality copy paper (higher quality paper contains watermarks that will

show up in the final image). It is essential that both the flat and the gel images be

identical, including the aperture, zoom (if applicable) and focus settings on the lens.

Step-by-Step Flat Field Calibration:

1) Place the gel or other application in the cabinet or dark room.

2) Adjust the aperture, zoom (if applicable) and focus on the lens.

3) Use auto-expose set to the normal selection and acquire an image of the gel.

(Alternatively, it is possible to select show saturation and then use expose

preview and adjust the exposure time manually to just under saturation.)

4) Save the image of the gel.

5) Next remove the gel from the UV transilluminator or white light tray and clean

and/or dry off the surface if necessary using glass cleaner.

6) Place the white piece of paper onto the appropriate surface. (For example, if

the UV transilluminator was used, place the paper onto the UV

transilluminator; if the white light tray was used, place the piece of paper onto

the white light tray.)

7) Turn on the appropriate light source used (white light, UV transilluminator, epi

lights, etc.).

8) Without changing anything on the lens acquire another image of the ‘Flat’

image following step #3 again.

9) Save the Flat image.

10) Open the original gel or other application image.

11) Select Flat Field Calibrate from the Image pull down menu.

12) Browse the directories for the ‘Flat’ image created.

13) Click on open. Make sure to save the flat field calibrated image for future use.

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Mitsubishi P91DW User Manual Page 57