Standard Operating Procedure - BioTek® Epoch™ 2 Microplate Spectrophotometer

Fisher Scientific S/N 1712184

Startup
1. Sign into the Epoch™2 logbook with your name, lab or PI, date, plate type (ie. 96-well, 384-
well etc.), and start time.
2. Turn on the touchscreen power button located on the top left of the screen.
3. Turn the power switch located on the bottom of the mouse to the on position.
4. Locate the power on/off switch on the front of the instrument, next to the carrier eject button
and turn to the on position. The microplate carrier will open briefly then close. The LED button
will flash green as the reader performs a power-up system test and there will be a slight
‘buzzing’ noise. DO NOT DISTURB the instrument during this time. Once complete, the reader
will extend the microplate carrier. Press the carrier eject button located on the bottom right hand
side of the instrument to close the microplate carrier until ready to use.
Using the Software and Setting up a protocol – 96 well or half area plates
1. The Gen5 TS v2.09 software will automatically be launched and the Task Manager window
will open.
2. Select desired protocol, create new or select read now.
3. The read now option located in the task manager is used to perform a quick read when no
other changes to a protocol have to be made.
Absorbance readings
1. To change the absorbance (OD) value, under the procedure tab at the top of the screen (this is
the ‘go-to’ tab to make most protocol changes) click on the ‘read’ option under the action
heading then click ok. You may select any absorbance value(s) between 200-999nm (± 2nm) –
you can select up to 6 different wavelengths to measure.
2. Under this tab you can also change the read type between endpoint, area scanning or spectral
scanning. You can also change the options for temperature, shake mode or modify kinetic
options.
3. To change the sample layout (what wells are read), click on ‘plate layout’ at the top of the
screen. You can ‘add’ or ‘delete’ well types here as needed.
4. Select desired well type between assay control, standard curve or sample and once complete
select ‘ok’.
Programmed OD600 – Bacterial Induction
1. Once the protocol is selected, the well selection window will open
BLK BLK BLK SPL SPL SPL SPL
(BLK – Blank. SPL – Sample)
2. Hold down the left button on the mouse to highlight the desired well and click ‘ok’.
3. The plate reader will eject. Gently place the 96 well plate with the A1 well located at the top
right then click ‘ok’.
4. The plate reader will close and automatically begin reading. Once complete, the microplate
carrier will eject and remove your plate. Close by pressing the carrier eject button.
Programmed OD595 – Bradford Assay
1. Once the protocol is selected, the well selection window will open
STD STD STD STD STD STD STD STD STD STD STD STD
STD STD STD STD STD STD STD STD STD STD STD STD
SPL SPL SPL SPL SPL SPL SPL SPL SPL SPL SPL SPL
 SPL SPL SPL SPL SPL SPL
(STD* – Standard. SPL – Sample)
These Standard concentrations have already been input into the program
2. Hold down the left button on the mouse to highlight the desired well and click ‘ok’.
3. The plate reader will eject. Gently place the 96 well plate with the A1 well located at the top
right then click ‘ok’.
4. The plate reader will close and automatically begin reading. Once complete, the microplate
carrier will eject and remove your plate. Close by pressing the carrier eject button.
Export a file - notepad
1. Click on the export button located at the top right of the screen
2. An ERROR message will appear but ignore it by clicking ‘ok’.
3. Save your file under the appropriate folder on the desktop.
Take3 Multi-volume plate
1. For measurements with very low (2 ul) sample volume, or to use with the BioCell and/or
stoppered Cuvettes.
2. Open the Gen5 TS v.2.09 software. The task manager will open and select the desired
application under the TAKE3 heading. The pre-programmed applications are:
Nucleic Acid Quantification – dsDNA, RNA or ssDNA (detection limit ~ 2ng/ul dsDNA
typical)
Protein A280 – BSA, IgG, Lysozyme
3. To access the microspots (16), gently open the lid to expose the glass slide (shown below).
4. Remove the protective foam covering. Pipet the samples (2 ul each) into the desired locations
and gently close the lid to avoid splashing. Read the plate immediately.

5. Place the plate in the microplate carrier by aligning the A1 position at the top right with the A1
position on the carrier.
6. Press the carrier eject button to close the microplate carrier.
7. In the Gen5 Take3 module, select the empty wells, blank wells or sample well locations by
using the drop-down menu and clicking on the location of the wells on the microspot image.
8. Select the samples to be read and click on ‘read’.
9. Once the read is complete, scroll the mouse over the well locations to indicate the A260, A280
and concentrations.
10. Once complete, the microplate carrier will eject and remove your plate. Close by pressing
the carrier eject button. Gently wipe off the microspots using a kim wipe. Replace the
protective foam covering and place back into the carrying case.
Shutdown
1. Ensure that you have REMOVED your sample from the microplate carrier and that it is
CLOSED.
2. Exit out of the Gen5 TS v2.09 software but clicking on the red X button on the right-hand side
of the screen.
3. Power down the machine by swiping left on the touch screen to bring up the main menu, tap
on the window icon and select log-out. The screen will go black. Swipe up on the main screen
and at the bottom right on the screen, tap on the power icon and select shut down. Cover the
microplate reader when complete.
4. If the Take3 plate was used, gently ensure to blot the fluid from both sides of the plate and use
another wipe to clean up any remaining fluid.
5. Replace the protective foam sheet and gently close the lid. Store the Take3 plate back into the
case.
6. If the touch screen was used on the reader, ensure that you have GENTLY wiped it off with
ONLY the provided BioTek® cleaning cloth.
7. Cover the instrument once all shutdown steps have been completed.
8. Turn the power switch located on the bottom of the mouse to the off position.
9. Sign out of the logbook with the time completed.
*** ! IMPORTANT ! ***
If ANY errors or issues are encountered prior to or during instrument use, you must FIRST
IMMEDIATELY inform your GAs and instructor or PI. They will then get in contact with the
Departmental Technician
Biochemistry Departmental Technician Contact Information
Joe Leicaa Email: jlichaa@uwindsor.ca
Phone: ext.4238 Office: 385-3 Essex Hall