GE InCell Analyzer 2500 HS - 1: Start up and perform a screen

Log-in with your ZMB credentials.

Start the "IN Cell Analyzer" software via the desktop icon.

In the cabinet close to the entrance door you can find the accessories including different sample holders.

Either use a well plate or choose a suitable sample holder.

In the software click on "Eject" to open the microscope door.

Insert your plate or sample holder.

Make sure that the orientation is correct (A1 should be on the top left corner).

In the software click on "Load".

Select a plate/slide template.

Select an objective.

Choose a suitable polychroic:

BGOFR_1: Blue (390/18), Green (475/28), Orange (524/27), Far Red (632/22)

BGOFR_2: Blue (390/18), Green (475/28), Red (575/25), Far Red (632/22)

Add the wanted channels by using the "+" icon.

Hoovering with the mouse over a channel opens a context information box with additional information.

To move the stage, click in the center of a well or cover slip.

Activate the "Laser Autofocus". The needed power level depends on the objective as well as the sample.

Check the laser auto focus trace by clicking "Verify":

If the signal is saturated, reduce the auto focus laser power.

If the "Measured Parameters" deviate more than 10% of the "Protocal/Plate Parameters" adjust the plate parameters.

Click "Apply Measured Parameters" for doing that.

Click "Apply" in the "Plate Editor" for adjusting the plate parameters to the measured values.

Check the "Software Autofocus" checkmark to engage the software autofocus.

Specify a z axis search range.

Define the channel as well as the frequency for performing a software auto focus.

Click on "Auto Offset" to automatically measure the offset for the selected channels.

When done, the software shows you the resulting offsets. They should be in a similar range for all channel. If a channel shows a quite different value then you can:

Move to another position and try again.

Check the signal in this channel, adjust the exposure (see next step) and try again.

Manually adjust the offset to the same value as in another channel.

The measured offsets will be set in the "AF Offset" window. They also can be manually adjusted there.

Select the first channel and press "AF". A laser auto focus will be performed, the offset applied and an image generated.

Check the image statistics. This microscope has a 16 Bit scmos camera so the dynamic range can be very high.

Depending on your statistics, adjust the exposure time.

Click on the camera symbol icon to make an image with the new exposure setting. When using the camera symbol no auto focus is performed.

Check again your image statistics.

Repeat this step for all channels.

In the left menu select the "Fields" settings.

Here you can select the number of fields acquired in each well.

Next you can adjust the field placement, spacing as well as the acquisition strategy for the fields in a well.

In the plate view you can select/unselect wells for acquisition.

Shift and holding the left mouse button can be used to unselect wells.

Control and holding the left mouse button can be used to select wells.

Before running a screen, you have to save your modified acquisition protocol.

You can give an additional experiment/plate ID which is used for naming the folders.

Specify a storage location for your image data. We recommend saving on DataMover (X:). You can later on move your data to the ZMB data network drive via the DataMover (see also the corresponding guide).

Click on "Scan" to start your scan.

For some imaging modalities such as when using deconvolution, it is necessary to reopen the dataset after acquisition and automatic processing.

Here you can change the displayed parameters/preview images for the plate overview.

Select a field for its data being displayed.

Explore the channels by adjusting the image intensities.

"Eject" and remove your sample. Click again on "Load" to close the microscope door.

Close the InCell software.

"Sign out" from the computer.