Olympus Microscope Manual

General rules:

Never use microscope without training

If you want to use the microscope for more than 2 hours: make a
reservation (list available on bioteclab dropbox)

Contact:

Glenn.vandestaey@cit.kuleuven.be

Kim.vekemans@cit.kuleuven.be

Rob.vandenbroeck@cit.kuleuven.be

1. Sample preparation
Apply three droplets of your sample on a sample glass, as shown in figure 1. Drop
a cover glass on each droplet, while taking care that you only use one cover
glass! Attach the cover glass to the sample glass using nail polish at the borders
(see figure 2).
Wait for the nail polish to dry. When dried, insert the sample on the sample stage
of the microscope with the cover glasses downwards (which is why the cover
glasses are attached with nail polish).

2. Starting up procedure

Turn everything on in the right order:

1. First, turn on the Control Box IX3-CBH (situated on the left hand side of the
computer),
2. then the touch panel controller (located on the left hand side of the
microscope itself) by pressing the on/off button on the backside,
3. and lastly, turn on the computer and the computer screen itself.

Choose the user IX83-user and insert the password obtained during the
training.

Open cellSens Dimension by double clicking the icon on the desktop, as

shown in Figure 1.

Figure 1: cellSens Dimension

Calibrate the stage: always click on ok or next

3. Closing procedure

Remove all stored images from the S-drive. The solid state drive is only used
for quick measuring, not for storage!

Turn everything off in the right order:

1. Turn off the computer and the computer screen,
2. then turn off the touch panel controller,
3. and lastly turn off the Control Box IX3-CBH.

To protect the objects against dust, it is important to cover the objective hole
in the stage!

4. Interface Overview
Error: Reference source not found shows the start-up interface of cellSens
Dimension. The major functions, illustrated with text boxes, will be discussed in
detail in the following subsections.

Figure 1: Interface Overview

4.1.

Camera Control (and Acquisition Settings)

An overview of the top part of the Camera Control Panel is shown in Figure 1.

Figure 1: Camera Control Panel

Click on the icon for Acquisition Settings, displayed in the top left corner as
part of the Camera Control Panel (see Figure 1).

Figure 1: Acquisition Settings

a. Expand Saving; 3 possibilities (see Figure 1)

For Snapshot:

Choose a file type (e.g. vsi or tiff). The vsi extension saves
all information, but can only be opened and manipulated
in the cellSens Dimension program. Tiff on the other hand
is the best option for further analysis using Matlab.

Change the Directory Path to the folder where you need

your images to be saved. This is very important: your
images will otherwise be saved in somebody elses folder.

For Movie and for Process/Experiment: the file type of the

obtained image/movie is always vsi/avi (if you need another
extension, change this manually afterwards). Like with Snapshot,
the directory path needs to be adjusted so that the images will
be saved in the right folder.

Figure 1: Acquisition Settings - Saving

b. Expand Camera and go to General. See that both boxes for horizontal
and vertical mirroring are checked, as shown in Figure 1.

Figure 1: Acquisition Settings - Camera

Live/Snap/Movie recording
a. Click on Live if you want to see what the microscope is seeing that
exact moment and place (this will appear in the middle of the screen),

b. click on Snap if you want to take a snap shot of that specific moment
at that specific place,
c. and check the box on Movie recording if you want to make a video.

The program can adjust the exposure time automatically or you can choose
one manually, as can be seen on Error: Reference source not found. Note that
'eyepieces,camera,50/50' are adjusted under Microscope Control.

The Auto-Focus function can literally be used to automatically bring the image
in focus. If not all objects are in the same plane however, this function
randomly chooses one object to focus upon (which causes the other objects to
be out of focus).

It is possible to take pictures in grey scale and in color, using the Toggle
RGB/Greyscale mode'-button.

4.2.

Microscope Control

A detailed overview of the Microscope Control Panel is given in Figure 1.

You can choose from 5 objectives: 4x, 10x, 20x, 40x, or 100x. The 100x
objective is an oil immersed lens, therefore you need to use oil immersed
samples and clean the microscope with a tissue after use.

As Observation Methods, it is possible to choose fluorescence (options: Blue

Fluorescent Protein, Green Fluorescent Protein, Texas Red, Yellow Fluorescent
Protein, Blue Fluorescent Protein Cyan) or transmission (options: Bright Field,
POLarised light, Phase contrast).
Extra:

Under Device Units, you can specify the Left-Port Changer:

a. choose Eyepieces if you want to observe the sample through the eyepieces,
b. choose Camera if you want to observe the sample through the camera
(shown in the cellSens Dimension program) and not directly, and
c. choose 50/50 if you want to observe the sample in both ways; through
the eye-pieces and through the camera.
If you choose the last option, be sure to turn of the light in the room to
minimize unwanted reflections.

Figure 1: Microscope Control Panel

4.3.

Stage Navigator

4.4.

Process Manager

5. Activated Sludge Imaging