Hidex Plate CHAMELEON V (quick start guide) · Turning ON 1. Turn on the computer 2. Turn on the power source (big black box behind luminometer) 3. Turn on the luminometer · MikroWin 2000 1. Start MikroWin (doubleclick on MikroWin 2000 icon on desktop) 2. Join MikroWin to luminometer - Read --> Options, choose Chameleon in Device field and press Ok · Setting up the measurement 1. Click on the icon ( ) next to file name in the lower left part of the screen to open the option screen 2. In the lower right part of the option screen choose which wells do you want to measure - black dots are selected wells, and if you want to measure by columns or by rows 3. In the lower left part of the option screen you can activate Temperature Unit and set the targeted temperature (2 - 42 °C), !!!the machine starts to heat up after starting any measurement, so if you wish to heat it up just run any random measurement first!!!, luminometer can not cool down, only heat 4. Click on Add Tech icon and choose Luminescence (other two possibilities does not work on our machine) 5. Click on Edit Tech icon to set up parameters of your measurements, for basic measurements use following set up: § In Measurement Mode screen set Operation Mode to Single Measurement if you want to measure your wells just once or Range Kinetic if you want to measure continuous measurement - set Interval Time equal to the number of wells you want to measure (10 wells = 10 sec IT), Number of Repeats depends on how long you want to measure and § how many wells you selected, if you want to measure one hour 10 wells then the NoR would be 360 (3600/10) - measurement will be a little bit longer than one hour but it does not matter § In the Shaker Unit screen you can Activate Shaker if you need it - Shake Mode enable you to set when do you want to shake, usually Ranges are used for shaking before every plate cycle, set the Shake Time and Shake Amplitude - for 96 well microtitre plate use 2 - 3 interval (the number means length in mm which the plate will be moved - bigger number means longer distance but slower shaking and the other way around) § click Ok § If you need more sophisticated settings (Measurement mode, shaker, dispenser...), you can set up more parameters clicking on Edit Item icon · Starting measurement 1. Type the name of the measurement to the File Name column and press start icon 2. Click on no when it ask you to set the parameters as default 3. After luminometer opens the lid, place your plate inside and push start · During measurement 1. You can see the exact luminescence numbers (in CPS) for each well when choosing Measurement Data on left side column in basic screen 2. You can also see curves for each well or for selected wells when choosing Kinetics or Selection, to see the curves you need to set up the Y axis range by clicking on icon placed in upper left side of the screen to appropriate number (when the values you are measuring are about 1000 CPS - set Y axis range to e.g. 2000), in selection mode you need to select wells you want to see by clicking on them - little red triangle shows selected well · Closing measurement 1. Wait until the measurement ends, lid will be automatically open, take away your plate and click proceed (the lid will be automatically closed), if you were heating the machine up choose if you want to keep heating or not, the data will be saved in the data folder by a name you chose before measurement 2. If you wish to stop measurement before it ends automatically just click on cancel icon on the bottom of the screen and then choose what do you want to do , usually click on end counting and the rest is the same like in the previous section · Extracting data 1. To extract your data in the *.txt file which can be opened in MS Excel click on Export icon in the upper side of the main screen, choose Raw Data Export Driver in the Active Export Driver field and press ok, then you will find your file in the Transfer folder, its shortcut can be easily found on the computers desktop, if you wish to save your file somewhere else, just set up a destination you want · Turning OFF 1. Check if there is no plate presented in the luminometer chamber, if not take it out! (Instrument--> Open lid/Close lid) 2. Close MikroWin software and computer 3. Turn off the luminometer 4. Turn off the power source PS.: If you wish to use dispenser or set more sophisticated options for measurements, please contact dr. Vojtek, Dobeš (room 116) or dr. Hyršl (room 113) and they will kindly help you.