Jones Lab:Protocols:Kup

Luciferase imaging

To capture data

1) Focus camera in brightfield mode
I do this in the dark, with the door open. Remember to set gains high!

2) Click 'Run TSI' from Circadian toolbar

3) Change settings as appropriate- time is in seconds
Remember to save the TRI file, set ND filter to 100% (ie. no filter)

4) Once data collection is complete load TRI from menu
Click first file in sequence

5) Check overview of data using CRT
TRI -> Show Count Rate Trend

To create a sequence file

1) Click on 'Load frames' on Circadian toolbar

2) Choose first image in folder

3) Open into a buffer

4) Doubleclick to open buffer

To annotate image with OAPs

Right click to cancel at any time

1) Click 'Circle OAP'

2) Tap on image to add OAP

3) Resize to suit

4) Click to confirm size

5) Click to confirm position

6) Label

7) Copy using copy command

To extract data

1) Click 'Extract OAP' command on circadian toolbar

Make sure selection = all frames (eg. selection 1-48)

2) Export to Excel